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1.
Chemosphere ; 361: 142487, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38821129

RESUMO

This study unveils the detoxification potential of insecticide-tolerant plant beneficial bacteria (PBB), i.e., Ciceribacter azotifigens SF1 and Serratia marcescens SRB1, in spinach treated with fipronil (FIP), profenofos (PF) and chlorantraniliprole (CLP) insecticides. Increasing insecticide doses (25-400 µg kg-1 soil) significantly curtailed germination attributes and growth of spinach cultivated at both bench-scale and in greenhouse experiments. Profenofos at 400 µg kg-1 exhibited maximum inhibitory effects and reduced germination by 55%; root and shoot length by 78% and 81%, respectively; dry matter accumulation in roots and shoots by 79% and 62%, respectively; leaf number by 87% and leaf area by 56%. Insecticide application caused morphological distortion in root tips/surfaces, increased levels of oxidative stress, and cell death in spinach. Application of insecticide-tolerant SF1 and SRB1 strains relieved insecticide pressure resulting in overall improvement in growth and physiology of spinach grown under insecticide stress. Ciceribacter azotifigens improved germination rate (10%); root biomass (53%); shoot biomass (25%); leaf area (10%); Chl-a (45%), Chl-b (36%) and carotenoid (48%) contents of spinach at 25 µg CLP kg-1 soil. PBB inoculation reinvigorated the stressed spinach and modulated the synthesis of phytochemicals, proline, malondialdehyde (MDA), superoxide anions (O2•-), and hydrogen peroxide (H2O2). Scanning electron microscopy (SEM) revealed recovery in root tip morphology and stomatal openings on abaxial leaf surfaces of PBB-inoculated spinach grown with insecticides. Ciceribacter azotifigens inoculation significantly increased intrinsic water use efficiency, transpiration rate, vapor pressure deficit, intracellular CO2 concentration, photosynthetic rate, and stomatal conductance in spinach exposed to 25 µg FIP kg-1. Also, C. azotifigens and S. marcescens modulated the antioxidant defense systems of insecticide-treated spinach. Bacterial strains were strongly colonized to root surfaces of insecticide-stressed spinach seedlings as revealed under SEM. The identification of insecticide-tolerant PBBs such as C. azotifigens and S. marcescens hold the potential for alleviating abiotic stress to spinach, thereby fostering enhanced and safe production within polluted agroecosystems.


Assuntos
Antioxidantes , Inseticidas , Folhas de Planta , Raízes de Plantas , Serratia marcescens , Poluentes do Solo , Spinacia oleracea , Spinacia oleracea/efeitos dos fármacos , Spinacia oleracea/fisiologia , Spinacia oleracea/metabolismo , Poluentes do Solo/toxicidade , Poluentes do Solo/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/microbiologia , Folhas de Planta/efeitos dos fármacos , Serratia marcescens/fisiologia , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/metabolismo , Antioxidantes/metabolismo , Inseticidas/toxicidade , Praguicidas/metabolismo , Praguicidas/toxicidade , Biodegradação Ambiental , Estresse Oxidativo/efeitos dos fármacos , Bacillaceae/metabolismo , Bacillaceae/fisiologia , Fotossíntese/efeitos dos fármacos , Microbiologia do Solo , Solo/química , Germinação/efeitos dos fármacos
2.
Biotechnol Lett ; 43(12): 2243-2257, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34652635

RESUMO

The biopreservation strategy allows extending the shelf life and food safety through the use of indigenous or controlled microbiota and their antimicrobial compounds. The aim of this work was to characterize an inhibitory substance with bacteriocin-like activity (Sak-59) produced by the potentially probiotic L. sakei strain from artisanal traditional Kazakh horse meat product Kazy. The maximum production of Sak-59 occurred at the stationary phase of the L. sakei growth. Sak-59 showed inhibitory activity against gram-positive meat spoilage bacteria strains of Listeria monocytogenes, Staphylococcus aureus, and pathogenic gram-negative bacteria strains of Serratia marcescens and Escherichia coli, but not against the tested Lactobacilli strains. Sak-59 activity, as measured by diffusion assay in agar wells, was completely suppressed after treatment with proteolytic enzymes and remained stable after treatment with α-amylase and lipase, indicating that Sak-59 is a peptide and most likely not glycosylated or lipidated. It was concluded that Sak-59 is a potential new bacteriocin with a characteristic activity spectrum, which can be useful in the food and feed industries.


Assuntos
Bacteriocinas/genética , Microbiologia de Alimentos , Latilactobacillus sakei/química , Produtos da Carne/microbiologia , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Bacteriocinas/isolamento & purificação , Bacteriocinas/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/patogenicidade , Armazenamento de Alimentos , Cavalos/microbiologia , Humanos , Latilactobacillus sakei/genética , Peptídeos/química , Peptídeos/farmacologia , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/patogenicidade , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/patogenicidade
3.
Biomolecules ; 11(2)2021 01 30.
Artigo em Inglês | MEDLINE | ID: mdl-33573343

RESUMO

Green synthesis of metal nanoparticles using plant extracts as capping and reducing agents for the biomedical applications has received considerable attention. Moreover, emergence and spread of multidrug resistance among bacterial pathogens has become a major health concern and lookout for novel alternative effective drugs has gained momentum. In current study, we synthesized gold nanoparticles using the seed extract of Trachyspermum ammi (TA-AuNPs), assessed its efficacy against drug resistant biofilms of Listeria monocytogenes and Serratia marcescens, and evaluated its anticancer potential against HepG2 cancer cell lines. Microwave-assisted green synthesis of gold nanoparticles was carried out and characterization was done using UV-vis spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), and dynamic light scattering (DLS). Most nanoparticles were observed as spherical and spheroidal with few anisotropies with an average crystalline size of 16.63 nm. Synthesized TA-AuNPs demonstrated significant biofilm inhibitory activity against L. monocytogenes (73%) as well as S. marcescens (81%). Exopolysaccharide (EPS), motility, and CSH, key elements that facilitate the formation and maintenance of biofilm were also inhibited significantly at the tested sub-minimum inhibitory concentrations (sub-MICs). Further, TA-AuNPs effectively obliterated preformed mature biofilms of S. marcescens and L. monocytogenes by 64% and 58%, respectively. Induction of intracellular ROS production in TA-AuNPs treated bacterial cells could be the plausible mechanism for the reduced biofilm formation in test pathogens. Administration of TA-AuNPs resulted in the arrest of cellular proliferation in a concentration-dependent manner. TA-AuNPs decrease the intracellular GSH in HepG2 cancer cell lines, cells become more prone to ROS generation, hence induce apoptosis. Thus, this work proposes a new eco-friendly and rapid approach for fabricating NPs which can be exploited for multifarious biomedical applications.


Assuntos
Antineoplásicos/farmacologia , Apiaceae/metabolismo , Ouro/química , Nanopartículas Metálicas/química , Espécies Reativas de Oxigênio , Sementes/metabolismo , Anisotropia , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Sobrevivência Celular , Glutationa Transferase/metabolismo , Química Verde , Células Hep G2 , Humanos , Luz , Peroxidação de Lipídeos , Listeria monocytogenes/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Transmissão , Micro-Ondas , Extratos Vegetais/farmacologia , Polissacarídeos Bacterianos/química , Espalhamento de Radiação , Serratia marcescens/efeitos dos fármacos , Sais de Tetrazólio/química , Tiazóis/química , Difração de Raios X
4.
Microb Drug Resist ; 27(8): 1105-1116, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33439767

RESUMO

Aim: To evaluate the antimicrobial and antibiofilm effects of chelerythrine (CHE) against carbapenem-resistant Serratia marcescens (CRSM). Materials and Methods: The minimum inhibitory concentration (MIC) of CHE against CRSM was determined using the agar dilution method. Changes in intracellular adenosine triphosphate (ATP) concentration, intracellular pH, cell membrane potential, and cell membrane integrity were investigated to assess the influence of CHE on the cell membrane. The effects of CHE on cell morphology were observed by field emission scanning electron microscopy (FESEM) and transmission electron microscopy. The antibiofilm formation of CHE was measured by crystal violet staining and visualized with confocal laser scanning microscopy (CLSM) and FESEM. The influence of CHE on biofilm components was further investigated using CLSM specific combined with double-dyeing methods. Results: Our results showed that CHE had an MIC at 125 µg/mL against CRSM was capable of inhibiting the growth of CRSM and destroying its cell membrane integrity, as well as obviously changing the cell morphology. Sub-MIC CHE displayed robust inhibitory effects against CRSM biofilm formation by mediating the production of biofilm components. In addition, CLSM- and FESEM-mediated evaluation of the damage of biofilm cells and biofilm persistence revealed that at high concentrations, CHE could compromise the cells within biofilms and remove preformed biofilms. Conclusion: CHE shows promise as a natural antimicrobial substance against biofilm-positive CRSM, with the potential to serve as an alternative therapeutic agent.


Assuntos
Antibacterianos/farmacologia , Benzofenantridinas/farmacologia , Biofilmes/efeitos dos fármacos , Enterobacteriáceas Resistentes a Carbapenêmicos/efeitos dos fármacos , Serratia marcescens/efeitos dos fármacos , Membrana Celular/fisiologia , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana
5.
Drug Deliv ; 27(1): 1271-1282, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32885688

RESUMO

Antibiotic resistance amongst microbial pathogens is a mounting serious issue in researchers and physicians. Various alternatives to overcome the multidrug-resistant bacterial infections are under search, and biofilm growth inhibition is one of them. In this investigation, a polymeric drug delivery system loaded with multi-serratial drugs to improve the delivery of drugs against urinary tract infection causative Serratia marcescens. The chitosan grafted pyromellitic dianhydride - cysteine (CS-g-PMDA-CYS) was conjugated with AuNPs by using the -SH group of CYS and RF (rifampicin) and INH (isoniazid) were loaded in AuNPs-fused CS-g-PMDA-CYS system. Several physicochemical techniques characterized this fabricated AuNPs/RF/INH/CS-g-PMDA-CYS system. The successful encapsulation of RF and INH in AuNPs-fused CS-g-PMDA-CYS polymer had confirmed, and it observed the loading capacity for RF and INH was 9.02% and 13.12%, respectively. The in vitro drug discharge pattern was perceived high in pH 5.5 compared with pH 7.4. The AuNPs/RF/INH/CS-g-PMDA-CYS escalates 74% of Caenorhabditis elegans survival during Serratia marcescens infection by aiming biofilm development and virulence in S. marcescens. Author postulate that the fabricated system is a promising drug carrier and delivery system for inhibition of multidrug-resistant bacterias like S. marcescens.


Assuntos
Antibacterianos/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Compostos de Ouro/administração & dosagem , Nanopartículas Metálicas/administração & dosagem , Serratia marcescens/efeitos dos fármacos , Animais , Antibacterianos/química , Benzoatos/administração & dosagem , Benzoatos/síntese química , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/microbiologia , Caenorhabditis elegans/fisiologia , Quitosana/administração & dosagem , Quitosana/síntese química , Cisteína/administração & dosagem , Cisteína/síntese química , Farmacorresistência Bacteriana Múltipla/fisiologia , Compostos de Ouro/síntese química , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana/métodos , Infecções por Serratia/tratamento farmacológico , Serratia marcescens/fisiologia , Infecções Urinárias/tratamento farmacológico , Difração de Raios X/métodos
6.
PLoS One ; 15(7): e0236505, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32701970

RESUMO

Multidrug resistance prompts the search for new sources of antibiotics with new targets at bacteria cell. To investigate the antibacterial activity of Cinnamomum cassia L. essential oil (CCeo) alone and in combination with antibiotics against carbapenemase-producing Klebsiella pneumoniae and Serratia marcescens. The antimicrobial susceptibility of the strains was determined by Vitek® 2 and confirmed by MALDI-TOF/TOF. The antibacterial activity of CCeo and its synergism with antibiotics was determined using agar disk diffusion, broth microdilution, time-kill, and checkboard methods. The integrity of the bacterial cell membrane in S. marcescens was monitored by protein leakage assay. CCeo exhibited inhibitory effects with MIC = 281.25 µg.mL-1. The association between CCeo and polymyxin B showed a decrease in terms of viable cell counts on survival curves over time after a 4 hour-treatment with a FIC index value of 0.006. Protein leakage was observed with increasing concentrations for CCeo and CCeo + polymyxin B treatments. CCeo showed antibacterial activity against the studied strains. When associated with polymyxin B, a synergistic effect was able to inhibit bacterial growth rapidly and consistently, making it a potential candidate for the development of an alternative treatment and drug delivery system for carbapenemase-producing strains.


Assuntos
Infecções por Klebsiella/tratamento farmacológico , Óleos Voláteis/farmacologia , Polimixina B/farmacologia , Infecções por Serratia/tratamento farmacológico , Antibacterianos/farmacologia , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/genética , Cinnamomum aromaticum/química , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla/genética , Sinergismo Farmacológico , Humanos , Infecções por Klebsiella/genética , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/patogenicidade , Infecções por Serratia/genética , Infecções por Serratia/microbiologia , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/patogenicidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , beta-Lactamases/genética
7.
Biofouling ; 36(3): 351-367, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32401555

RESUMO

Pseudomonas aeruginosa and Serratia marcescens are prominent members belonging to the group of ESKAPE pathogens responsible for Urinary Tract Infections (UTI) and nosocomial infections. Both the pathogens regulate several virulence factors, including biofilm formation through quorum sensing (QS), an intercellular communication mechanism. The present study describes the anti-biofilm and QS quenching effect of thiazolinyl-picolinamide based palladium(II) complexes against P. aeruginosa and S. marcescens. Palladium(II) complexes showed quorum sensing inhibitory potential in inhibiting swarming motility behaviour, pyocyanin production and other QS mediated virulence factors in both P. aeruginosa and S. marcescens. In addition, the establishment of biofilms was prevented on palladium (II) coated catheters. Overall, the present study demonstrates that thiazolinyl-picolinamide based palladium (II) complexes will be a promising strategy to combat device-mediated UTI infections.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Complexos de Coordenação/farmacologia , Paládio/farmacologia , Ácidos Picolínicos/química , Tiazóis/química , Cateteres Urinários/microbiologia , Antibacterianos/química , Antibacterianos/toxicidade , Biofilmes/crescimento & desenvolvimento , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/química , Complexos de Coordenação/toxicidade , Infecção Hospitalar/prevenção & controle , Humanos , Células MCF-7 , Paládio/química , Paládio/toxicidade , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Piocianina/metabolismo , Percepção de Quorum/efeitos dos fármacos , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/metabolismo , Infecções Urinárias/microbiologia , Infecções Urinárias/prevenção & controle , Virulência , Fatores de Virulência/metabolismo
8.
Sci Rep ; 10(1): 5279, 2020 03 24.
Artigo em Inglês | MEDLINE | ID: mdl-32210346

RESUMO

Melia azedarach-rhizosphere mediated degradation of benzo(a)pyrene (BaP), in the presence of cadmium (Cd) was studied, using efficient rhizobacterial isolate. Serratia marcescens S2I7, isolated from the petroleum-contaminated site, was able to tolerate up to 3.25 mM Cd. In the presence of Cd, the isolate S2I7 exhibited an increase in the activity of stress-responsive enzyme, glutathione-S-transferase. Gas Chromatography-Mass spectroscopy analysis revealed up to 59% in -vitro degradation of BaP after 21 days, while in the presence of Cd, the degradation was decreased by 14%. The bacterial isolate showed excellent plant growth-promoting attributes and could enhance the growth of host plant in Cd contaminated soil. The 52,41,555 bp genome of isolate S. marcescens S2I7 was sequenced, assembled and annotated into 4694 genes. Among these, 89 genes were identified for the metabolism of aromatic compounds and 172 genes for metal resistance, including the efflux pump system. A 2 MB segment of the genome was identified to contain operons for protocatechuate degradation, catechol degradation, benzoate degradation, and an IclR type regulatory protein pcaR, reported to be involved in the regulation of protocatechuate degradation. A pot trial was performed to validate the ability of S2I7 for rhizodegradation of BaP when applied through Melia azedarach rhizosphere. The rhizodegradation of BaP was significantly higher when augmented with S2I7 (85%) than degradation in bulk soil (68%), but decreased in the presence of Cd (71%).


Assuntos
Benzo(a)pireno/metabolismo , Biodegradação Ambiental/efeitos dos fármacos , Cádmio/toxicidade , Melia azedarach/efeitos dos fármacos , Rizosfera , Serratia marcescens/metabolismo , Microbiologia do Solo , Poluentes do Solo/toxicidade , Proteínas de Bactérias/metabolismo , Catecol 1,2-Dioxigenase/metabolismo , Catecol 2,3-Dioxigenase/metabolismo , Catecóis/metabolismo , DNA Bacteriano/genética , Cromatografia Gasosa-Espectrometria de Massas , Genoma Bacteriano , Glutationa Transferase/metabolismo , Hidroxibenzoatos/metabolismo , Melia azedarach/crescimento & desenvolvimento , Óperon , Filogenia , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/genética , Serratia marcescens/isolamento & purificação , Ácido Succínico/farmacologia
9.
Arch Insect Biochem Physiol ; 103(1): e21626, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31562754

RESUMO

Insects can produce various antimicrobial peptides (AMPs) upon immune stimulation. One class of AMPs are characterized by their high proline content in certain fragments. They are generally called proline-rich antimicrobial peptides (PrAMPs). We previously reported the characterization of Spodoptera litura lebocin-1 (SlLeb-1), a PrAMP proprotein. Preliminary studies with synthetic polypeptides showed that among the four deductive active fragments, the C-terminal fragment SlLeb-1 (124-158) showed strong antibacterial activities. Here, we further characterized the antibacterial and antifungal activities of 124-158 and its four subfragments: 124-155, 124-149, 127-158, and 135-158. Only 124-158 and 127-158 could agglutinate bacteria, while 124-158 and four subfragments all could agglutinate Beauveria bassiana spores. Confocal microscopy showed that fluorescent peptides were located on the microbial surface. Fragment 135-158 lost activity completely against Escherichia coli and Staphylococcus aureus, and partially against Bacillus subtilis. Only 124-149 showed low activity against Serratia marcescens. Negative staining, transmission, and scanning electron microscopy of 124-158 treated bacteria showed different morphologies. Flow cytometry analysis of S. aureus showed that 124-158 and four subfragments changed bacterial subpopulations and caused an increase of DNA content. These results indicate that active fragments of SlLeb-1 may have diverse antimicrobial effects against different microbes. This study may provide an insight into the development of novel antimicrobial agents.


Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Proteínas de Insetos/farmacologia , Spodoptera/química , Animais , Peptídeos Catiônicos Antimicrobianos/química , Bacillus subtilis/efeitos dos fármacos , Beauveria/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Proteínas de Insetos/química , Serratia marcescens/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos
10.
Rev Soc Bras Med Trop ; 52: e20180502, 2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31271619

RESUMO

INTRODUCTION: Plant products are sources for drug development against multidrug resistant bacteria. METHODS: The antimicrobial activity of Origanum vulgare L. essential oil (OVeo) against carbapenem-resistant strains was assessed by disk-diffusion, microdilution (REMA-Resazurin Microtiter Assay), and time kill assays. RESULTS: Carbapenemase production was confirmed for all strains. OVeo exhibited a minimum inhibitory concentration of 0.059% v/v for Klebsiella pneumoniae and Serratia marcescens, and of 0.015 % v/v for Acinetobacter baumannii. A decrease in cell count was observed after a 4 h treatment. CONCLUSIONS: OVeo antimicrobial effect was rapid and consistent, making it a candidate for developing alternative therapeutic options against carbapenem-resistant strains.


Assuntos
Acinetobacter baumannii/efeitos dos fármacos , Antibacterianos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Klebsiella pneumoniae/efeitos dos fármacos , Óleos Voláteis/farmacologia , Origanum/química , Serratia marcescens/efeitos dos fármacos , Acinetobacter baumannii/crescimento & desenvolvimento , Antibacterianos/classificação , Proteínas de Bactérias , Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana Múltipla , Bactérias Gram-Negativas/crescimento & desenvolvimento , Humanos , Klebsiella pneumoniae/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Serratia marcescens/crescimento & desenvolvimento , beta-Lactamases
11.
Carbohydr Polym ; 218: 243-249, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31221327

RESUMO

Natural cellulose fibers were extracted from a fast growing perennial grass Eulaliopsis binata (commonly known as Sabai) and characterized for their structure and properties. The untreated sabai grass has been used as reinforcement for polypropylene composites and properties of the composites have been investigated. Although the composition of the sabai grass is typical to other lignocellulosic sources, there is a high content of flavonoids (630 mg/g) and phenols (510 mg/g) which provides high antibacterial, and antifungal properties to the fibers and composites developed. Fiber bundles extracted from the grass had tensile strength of 493 MPa and tensile modulus of 21 GPa, similar to common natural cellulose fibers. Both tensile and flexural properties of polypropylene composites increased with increasing ratio of sabai grass. Polypropylene composites reinforced with sabai grass show high noise insulation and thermal resistance properties suggesting their suitability for automotive and building applications.


Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Celulose/farmacologia , Extratos Vegetais/farmacologia , Poaceae/química , Antibacterianos/química , Antifúngicos/química , Aspergillus niger/efeitos dos fármacos , Bacillus cereus/efeitos dos fármacos , Celulose/química , Cryptococcus/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Teste de Materiais , Extratos Vegetais/química , Maleabilidade , Polipropilenos/química , Serratia marcescens/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Resistência à Tração , Condutividade Térmica
12.
Mol Med Rep ; 20(1): 745-754, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31180518

RESUMO

The present study aimed to further clarify the genetic mechanisms responsible for the antimicrobial resistance of Serratia marcescens (S. marcescens) using RNA sequencing. Three drug­susceptible S. marcescens strains (named MYQT1, MYQT2, and MYQT3) and three multidrug­resistant S. marcescens strains (named MYQT4, MYQT5, and MYQT6) were isolated from six different patients and subjected to RNA sequencing. Differentially expressed genes (DEGs) between the multidrug­resistant S. marcescens strains and drug­susceptible strains were screened and compared, followed by functional enrichment analysis. In addition, a protein­protein interaction (PPI) network was constructed, and significant modules were extracted from it. Genes enriched in the significant modules were subjected to further enrichment analysis. MYQT3 had very a different expression pattern from MYQT1 and MYQT2, and thus, MYQT3 was excluded from the following analysis. A total of 225 DEGs were identified, of which SMDB11_RS09300 (GTP cyclohydrolase FolE2) was the most significantly upregulated with a log2 FC of 6.4; these DEGs were enriched in different GO terms, including hydrogen sulfide biosynthetic process, sulfur compound transmembrane transporter activity, and ABC transporter complex. Additionally, several genes were identified to be important genes in the PPI network, including SMDB11_RS17755 (upregulated; glutamate synthase large subunit), SMDB11_RS00590 (upregulated; sulfite reductase subunit α), and SMDB11_RS04505 (upregulated; cystathionine ß­synthase). Thus, SMDB11_RS09300, SMDB11_RS17755, SMDB11_RS00590, and SMDB11_RS04505 may play significant roles in the antimicrobial resistance of S. marcescens by participating in folate metabolism or the integrity of cell membranes. However, further experiments are required to clarify these findings.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/genética , Regulação Bacteriana da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Mapeamento de Interação de Proteínas , Infecções por Serratia/tratamento farmacológico , Infecções por Serratia/microbiologia , Serratia marcescens/metabolismo , Transcriptoma
13.
Cell Microbiol ; 21(9): e13045, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31099073

RESUMO

Serratia marcescens is an opportunistic pathogen with increasing incidence in clinical settings. This is mainly attributed to the timely expression of a wide diversity of virulence factors and intrinsic and acquired resistance to antibiotics, including ß-lactams, aminoglycosides, quinolones, and polypeptides. For these reasons, S. marcescens has been recently categorised by the World Health Organization as one priority to strengthen efforts directed to develop new antibacterial agents. Therefore, it becomes critical to understand the underlying mechanisms that allow Serratia to succeed within the host. S. marcescens ShlA pore-forming toxin mediates phenotypes that alter homeostatic and signal transduction pathways of host cells. It has been previously demonstrated that ShlA provokes cytotoxicity, haemolysis and autophagy and also directs Serratia egress and dissemination from invaded nonphagocytic cells. However, molecular details of ShlA mechanism of action are still not fully elucidated. In this work, we demonstrate that Ni2+ selectively and reversibly blocks ShlA action, turning wild-type S. marcescens into a shlA mutant strain phenocopy. Combined use of Ni2+ and calcium chelators allow to discern ShlA-triggered phenotypes that require intracellular calcium mobilisation and reveal ShlA function as a calcium channel, providing new insights into ShlA mode of action on target cells.


Assuntos
Proteínas de Bactérias/antagonistas & inibidores , Canais de Cálcio/metabolismo , Proteínas Hemolisinas/antagonistas & inibidores , Níquel/farmacologia , Serratia marcescens/efeitos dos fármacos , Fatores de Virulência/metabolismo , Animais , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/toxicidade , Toxinas Bacterianas/metabolismo , Células CHO , Cálcio/metabolismo , Cricetulus , Eritrócitos/microbiologia , Proteínas Hemolisinas/metabolismo , Proteínas Hemolisinas/toxicidade , Hemólise/efeitos dos fármacos , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Humanos , Fenótipo , Serratia marcescens/metabolismo , Serratia marcescens/patogenicidade
14.
Rev. Soc. Bras. Med. Trop ; 52: e20180502, 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1041563

RESUMO

Abstract INTRODUCTION: Plant products are sources for drug development against multidrug resistant bacteria. METHODS The antimicrobial activity of Origanum vulgare L. essential oil (OVeo) against carbapenem-resistant strains was assessed by disk-diffusion, microdilution (REMA-Resazurin Microtiter Assay), and time kill assays. RESULTS Carbapenemase production was confirmed for all strains. OVeo exhibited a minimum inhibitory concentration of 0.059% v/v for Klebsiella pneumoniae and Serratia marcescens, and of 0.015 % v/v for Acinetobacter baumannii. A decrease in cell count was observed after a 4 h treatment. CONCLUSIONS OVeo antimicrobial effect was rapid and consistent, making it a candidate for developing alternative therapeutic options against carbapenem-resistant strains.


Assuntos
Humanos , Serratia marcescens/efeitos dos fármacos , Óleos Voláteis/farmacologia , Acinetobacter baumannii/efeitos dos fármacos , Origanum/química , Bactérias Gram-Negativas/efeitos dos fármacos , Klebsiella pneumoniae/efeitos dos fármacos , Antibacterianos/farmacologia , Serratia marcescens/crescimento & desenvolvimento , Proteínas de Bactérias , beta-Lactamases , Testes de Sensibilidade Microbiana , Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana Múltipla , Acinetobacter baumannii/crescimento & desenvolvimento , Bactérias Gram-Negativas/crescimento & desenvolvimento , Klebsiella pneumoniae/crescimento & desenvolvimento , Antibacterianos/classificação
15.
Microb Pathog ; 125: 189-195, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30227230

RESUMO

This work illustrates a new role for the membranotropic peptide gH625 and its derivative gH625-GCGKKK in impairing formation of polymicrobial biofilms. Mixed biofilms composed of Candida and bacterial species cause frequently infections and failure of medical silicone devices and also show a major drug resistance than single-species biofilms. Inhibition and eradication of biofilms were evaluated by complementary methods: XTT-reduction, and crystal violet staining (CV). Our results indicate that gH625-GCGKKKK, better than the native peptide, strongly inhibited formation of mixed biofilms of clinical isolates of C. tropicalis/S. marcescens and C. tropicalis/S. aureus and reduced the biofilm architecture, interfering with cell adhesion and polymeric matrix, as well as eradicated the long-term polymicrobial biofilms on silicone surface.


Assuntos
Anti-Infecciosos/metabolismo , Biofilmes/efeitos dos fármacos , Candida tropicalis/efeitos dos fármacos , Peptídeos/metabolismo , Serratia marcescens/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Proteínas do Envelope Viral/metabolismo , Candida tropicalis/crescimento & desenvolvimento , Formazans/análise , Violeta Genciana/análise , Serratia marcescens/crescimento & desenvolvimento , Coloração e Rotulagem , Staphylococcus aureus/crescimento & desenvolvimento
16.
Antonie Van Leeuwenhoek ; 111(4): 501-515, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29101490

RESUMO

The aim of this study was to evaluate the anti-biofilm and anti-virulence properties of petroselinic acid (PSA) against the environmental pathogen Serratia marcescens. PSA significantly inhibited the quorum sensing (QS)-dependent virulence factors such as prodigiosin, protease productions, and biofilm formation in S. marcescens. The antibiofilm potential of PSA was also confirmed through light, confocal laser scanning, and scanning electron microscopic analyses. Furthermore, PSA effectively inhibited the biofilm-related phenomena such as exopolysaccharide production, hydrophobicity production, swimming, and swarming motility without affecting the bacterial growth. In FT-IR analysis, the PSA treated S. marcescens cells displayed a reduction in cellular components compared to the untreated controls. The real-time analysis revealed the downregulation of QS-controlled virulence genes such as bsmB, fimA, fimC, and flhD in S. marcescens on treatment with PSA. The obtained results strongly suggested that PSA could be further explored as an antipathogenic drug to treat QS-mediated infections caused by S. marcescens.


Assuntos
Biofilmes/efeitos dos fármacos , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Ácidos Oleicos/farmacologia , Percepção de Quorum/genética , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/fisiologia , Virulência/efeitos dos fármacos , Antibacterianos/farmacologia , Polissacarídeos Bacterianos/genética , Espectroscopia de Infravermelho com Transformada de Fourier , Virulência/genética , Fatores de Virulência/genética
17.
PLoS One ; 12(10): e0185984, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29016698

RESUMO

The impact of substituents on the photochemical and biological properties of tetraphenylporphyrin-based photosensitizers for photodynamic therapy of cancer (PDT) as well as photodynamic inactivation of microorganisms (PDI) was examined. Spectroscopic and physicochemical properties were related with therapeutic efficacy in PDT of cancer and PDI of microbial cells in vitro. Less polar halogenated, sulfonamide porphyrins were most readily taken up by cells compared to hydrophilic and anionic porphyrins. The uptake and PDT of a hydrophilic porphyrin was significantly enhanced with incorporation in polymeric micelles (Pluronic L121). Photodynamic inactivation studies were performed against Gram-positive (S. aureus, E. faecalis), Gram-negative bacteria (E. coli, P. aeruginosa, S. marcescens) and fungal yeast (C. albicans). We observed a 6 logs reduction of S. aureus after irradiation (10 J/cm2) in the presence of 20 µM of hydrophilic porphyrin, but this was not improved with incorporation in Pluronic L121. A 2-3 logs reduction was obtained for E. coli using similar doses, and a decrease of 3-4 logs was achieved for C. albicans. Rational substitution of tetraphenylporphyrins improves their photodynamic properties and informs on strategies to obtain photosensitizers for efficient PDT and PDI. However, the design of the photosensitizers must be accompanied by the development of tailored drug formulations.


Assuntos
Anti-Infecciosos/química , Antineoplásicos/química , Fármacos Fotossensibilizantes/química , Porfirinas/química , Anti-Infecciosos/síntese química , Anti-Infecciosos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Transporte Biológico , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Candida albicans/efeitos da radiação , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Desenho de Fármacos , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/crescimento & desenvolvimento , Enterococcus faecalis/efeitos da radiação , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/efeitos da radiação , Halogenação , Humanos , Interações Hidrofóbicas e Hidrofílicas , Luz , Micelas , Testes de Sensibilidade Microbiana , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/farmacologia , Poloxâmero/química , Porfirinas/síntese química , Porfirinas/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/efeitos da radiação , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/efeitos da radiação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/efeitos da radiação , Relação Estrutura-Atividade , Sulfonamidas/química
18.
Biopolymers ; 107(11)2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28925099

RESUMO

A series of amphiphilic diblock copolypeptides (K30 -b-F15 , K30 -b-F30 , and K30 -b-F45 ) were synthesized via N-carboxy-α-amino-anhydride ring-opening polymerization. The copolypeptides had excellent antibacterial efficacy to both Gram positive (S. aureus) and Gram negative (E. coli) bacteria. The minimum inhibitory concentrations (MICs) against E. coli and S. aureus are 8 µg mL-1 and 2 µg mL-1 , respectively, lower than most natural and artificial antimicrobial peptides (AMPs). The morphological changes of the bacteria treated with diblock copolypeptides were investigated by transmission electron microscopy; the results proved that the diblock copolypeptides had a similar antibacterial pore-forming mechanism to natural cationic peptides. This was confirmed by laser scanning confocal microscope images. CCK-8 results and the MICs showed that the diblock copolypeptides have high selectivity to bacteria, which suggested that the diblock copolypeptides could be excellent candidates to replace traditional antibiotics in future.


Assuntos
Antibacterianos/farmacologia , Peptídeos/química , Peptídeos/farmacologia , Antibacterianos/efeitos adversos , Antibacterianos/química , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos/métodos , Escherichia coli/efeitos dos fármacos , Humanos , Lisina/química , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Transmissão , Peptídeos/síntese química , Fenilalanina/química , Pseudomonas aeruginosa/efeitos dos fármacos , Serratia marcescens/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos
19.
Burns ; 43(6): 1189-1194, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28641915

RESUMO

INTRODUCTION/BACKGROUND: Xeroform® is a petrolatum-based fine mesh gauze containing 3% bismuth tribromophenate. Bismuth, similar to other metals, has antimicrobial properties. Xeroform® has been used for decades in burn and plastic surgery as a donor site dressing and as a covering for wounds or partial thickness burns. Despite this, the antimicrobial spectrum of Xeroform® remains largely unknown. We examined the in-vitro efficacy of Xeroform® against common burn pathogens using zone-of-inhibition methodology in a commercial research facility. METHODS/DESIGN: Pure strains of 15 common burn pathogens including Methicillin-resistant Staphylococcus aureus (MRSA), Methicillin-sensitive Staphylococcus aureus (MSSA), Staphylococcus epidermidis, Pseudomonas aeruginosa, Enterobacter cloacae, Escherichia coli, Candida albicans, Vancomycin resistant Enterococcus, Acinetobacter baumennii, Klebsiella pneumonia, Extended spectrum beta-lactamase producing Klebsiella, Beta hemolytic Streptococcus pyogenes, Proteus mirabilis, Serratia marcescens, and Salmonella enterica ssp. Enterica were inoculated at a strength of 106-1010 CFU/ml onto appropriate agar plates. A sterile 1 in2 Xeroform® square was placed in the center of each plate, and the Zone of Inhibition (ZOI) was measured following 18-24h of incubation at 37°C. A second bismuth pharmaceutical (bismuth subsalicylate, Pepto-Bismol®) was then tested using the same methodology against the same strains of MRSA, MSSA, E. coli, K. pneumonia and S. marcescens. Finally, 3% w/v bismuth tribromophenate in glycerol suspension was tested against 13 burn pathogens for antimicrobial activity independent of the Xeroform® dressing by measure of Zone of Inhibition. RESULTS/FINDINGS: For Xeroform®, none of the fifteen pathogens had a measurable zone of inhibition on any plate. Bismuth subsalicylate showed a zone of inhibition for MSSA in 3 plates (mean of 47.2mm), in one of three plates for MRSA (13.8mm), and in one of three plates for S. marcesens (89.6mm). There was no zone of inhibition seen for K. pneumonia or E. coli. Bismuth tribromophenate, when not bound to Xeroform® showed activity against 12 of 13 pathogens. CONCLUSIONS/IMPLICATIONS: While bismuth subsalicylate, and bismuth tribromophenate unbound to Xeroform® demonstrate antimicrobial activity, it appears that Xeroform® dressings do not. The utility of Xeroform® in burn medicine may relate more to use as an impervious dressing than to antimicrobial effect. Donor sites are clean surgical wounds and clean partial thickness burns may have minimal colonization present. In such circumstances, an inactive and impervious dressing may be all that is necessary to promote wound healing.


Assuntos
Anti-Infecciosos/farmacologia , Bactérias/efeitos dos fármacos , Queimaduras/microbiologia , Candida albicans/efeitos dos fármacos , Fenóis/farmacologia , Bandagens , Farmacorresistência Bacteriana , Enterobacter cloacae/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Humanos , Técnicas In Vitro , Klebsiella/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Proteus mirabilis/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Salmonella enterica/efeitos dos fármacos , Serratia marcescens/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Streptococcus pyogenes/efeitos dos fármacos , Enterococos Resistentes à Vancomicina/efeitos dos fármacos
20.
Eye Contact Lens ; 43(2): 110-115, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26974534

RESUMO

OBJECTIVES: Contact lens-acquired bacterial infections are a serious problem. Of the reported cases, inadequate cleaning of the lens case was the most common cause of lens contamination. Organoselenium has been shown to inhibit bacterial attachment to different polymer materials. This study evaluates the ability of an organoselenium monomer, incorporated into the polymer of a polypropylene contact lens case coupon, to block the formation of biofilms in a lens case. METHODS: The bacteria tested were Pseudomonas aeruginosa, Staphylococcus aureus, Stenotrophomonas maltophilia, and Serratia marcescens. For this study, the bacteria were allowed to grow overnight, in trypticase soy broth media, in the presence of the selenium-containing polymer or the same polymer without organoselenium. The material was studied by both colony-forming unit determination and by confocal laser scanning microscopy. RESULTS: The results showed that the organoselenium polymer versus the control polymer resulted in the following effect on biofilm formation: (1) a reduction in P. aeruginosa of 7.3 logs (100%); (2) a reduction in S. aureus of 7.3 logs (100%); (3) a reduction in S. maltophilia of 7.5 logs (100%); and (4) a reduction in S. marcescens reduction of 3.3 logs (99.9%). To test the stability of the organoselenium polypropylene contact lens coupon, the coupon was soaked in PBS for eight weeks at room temperature. It was found that when these soaked coupons were tested against S. aureus, complete inhibition (8.1 logs) was obtained. Because organoselenium cannot leach from the polymer, this would imply that the organoselenium polypropylene contact lens case coupon would be inhibitory toward bacterial biofilm for the life of the case. CONCLUSION: The organoselenium polypropylene contact lens case coupon shows the ability to inhibit biofilm formation. The use of organoselenium copolymer should play an important role in protecting against contact lens case-acquired infection.


Assuntos
Biofilmes/efeitos dos fármacos , Lentes de Contato/microbiologia , Contaminação de Equipamentos/prevenção & controle , Compostos Organosselênicos/farmacologia , Soluções para Lentes de Contato/farmacologia , Infecções Oculares Bacterianas/prevenção & controle , Humanos , Compostos Organosselênicos/química , Polipropilenos/química , Pseudomonas aeruginosa/efeitos dos fármacos , Serratia marcescens/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Stenotrophomonas maltophilia/efeitos dos fármacos
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