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1.
Molecules ; 27(4)2022 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-35209213

RESUMEN

The design of multifunctional microcarriers has attracted significant attention because they combine various functions within a single system. In this study, we developed a set of multilayered hydrogel microcarriers, which were first loaded with chemotherapeutic curcumin (CUR), then, using the layer-by-layer (LbL) technique, coated through a polyelectrolyte shell consisting of chitosan (CHIT) or poly(allylamine hydrochloride) (PAH). As an outer layer with antimicrobial function, newly synthesised alkylene quaternary ammonium salt functionalised polyelectrolytes (A-QAS-PEs) were applied. For this purpose, poly(acrylic acid) (PAA) was decorated with different hydrophobic side chains (n-hexane and n-dodecane side entities) and different degrees of substitution (m) of quaternary ammonium groups (abbreviated as PAA-C(O)O-(CH2)n-N+(CH3)3(m); n = 6, 12; m = 8-14%). The grafting approach of PAA with the alkylene quaternary ammonium salt moiety was performed under mild reaction conditions using Steglich esterification followed by quaternisation. The structure of antimicrobial decorated PAA was confirmed by 1H NMR and FTIR, and the mean diameter of all multifunctional microparticles was characterised by SEM. The viscoelastic properties of the functional layers were studied using quartz crystal microbalance with a dissipation (QCM-D). The release of CUR from the microcarriers was described using a hybrid model, i.e., a combination of first-order kinetics and the Korsmeyer-Peppas model. The antimicrobial activity of functionalised PAA and multilayered CUR-loaded hydrogel microcarriers with quaternary ammonium function was assessed against Staphylococcus aureus and Serratia marcescens by the agar diffusion assay method. Only a limited inhibition zone of PAA was observed, but in the case of both antimicrobial decorated PAA and the corresponding multilayered nanocarriers, the inhibitory activity increase was achieved against both strains of bacteria.


Asunto(s)
Antibacterianos , Curcumina , Portadores de Fármacos , Hidrogeles , Serratia marcescens/crecimiento & desarrollo , Staphylococcus aureus/crecimiento & desarrollo , Antibacterianos/química , Antibacterianos/farmacocinética , Antibacterianos/farmacología , Curcumina/química , Curcumina/farmacocinética , Curcumina/farmacología , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacología , Hidrogeles/química , Hidrogeles/farmacocinética , Hidrogeles/farmacología
2.
Appl Biochem Biotechnol ; 194(2): 671-693, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-34449042

RESUMEN

The growth of respiratory diseases, as witnessed through the SARS and COVID-19 outbreaks, and antimicrobial-resistance together pose a serious threat to humanity. One reason for antimicrobial resistance is formation of bacterial biofilms. In this study the sulphated polysaccharides from green algae Chlamydomonas reinhardtii (Cr-SPs) is tested for its antibacterial and antibiofilm potential against Klebsiella pneumoniae and Serratia marcescens. Agar cup assay clearly indicated the antibacterial potential of Cr-SPs. Minimum inhibitory concentration (MIC50) of Cr-SPs against Klebsiella pneumoniae was found to be 850 µg/ml, and it is 800 µg/ml in Serratia marcescens. Time-kill and colony-forming ability assays suggest the concentration-dependent bactericidal potential of Cr-SPs. Cr-SPs showed 74-100% decrease in biofilm formation in a concentration-dependent manner by modifying the cell surface hydrophobic properties of these bacteria. Cr-SPs have also distorted preformed-biofilms by their ability to interact and destroy the extra polymeric substance and eDNA of the matured biofilm. Scanning electron microscopy analysis showed that Cr-SPs effectively altered the morphology of these bacterial cells and distorted the bacterial biofilms. Furthermore reduced protease, urease and prodigiosin pigment production suggest that Cr-SPs interferes the quorum sensing mechanism in these bacteria. The current study paves way towards developing Cr-SPs as a control strategy for treatment of respiratory tract infections.


Asunto(s)
Biopelículas/efectos de los fármacos , Polisacáridos/farmacología , Percepción de Quorum/efectos de los fármacos , Infecciones del Sistema Respiratorio/tratamiento farmacológico , Antibacterianos/química , Antibacterianos/farmacología , Biopelículas/crecimiento & desarrollo , COVID-19/virología , Chlorophyta/química , Humanos , Klebsiella pneumoniae/crecimiento & desarrollo , Klebsiella pneumoniae/patogenicidad , Pruebas de Sensibilidad Microbiana , Polisacáridos/química , Infecciones del Sistema Respiratorio/microbiología , SARS-CoV-2/efectos de los fármacos , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/patogenicidad , Tratamiento Farmacológico de COVID-19
3.
Microbiol Spectr ; 9(2): e0039421, 2021 10 31.
Artículo en Inglés | MEDLINE | ID: mdl-34523998

RESUMEN

Commensal microbes in animal guts often help to exclude bacterial pathogens. In honey bees, perturbing or depleting the gut microbiota increases host mortality rates upon challenge with the opportunistic pathogen Serratia marcescens, suggesting antagonism between S. marcescens and one or more members of the bee gut microbiota. In laboratory culture, S. marcescens uses a type VI secretion system (T6SS) to kill bacterial competitors, but the role of this T6SS within hosts is unknown. Using infection assays, we determined how the microbiota impacts the abundance and persistence of S. marcescens in the gut and visualized colocalization of S. marcescens with specific community members in situ. Using T6SS-deficient S. marcescens strains, we measured T6SS-dependent killing of gut isolates in vitro and compared the persistence of mutant and wild-type strains in the gut. We found that S. marcescens is rapidly eliminated in the presence of the microbiota but persists in microbiota-free guts. Protection is reduced in monocolonized and antibiotic-treated bees, possibly because different symbionts occupy distinct niches. Serratia marcescens uses a T6SS to antagonize Escherichia coli and other S. marcescens strains but shows limited ability to kill bee symbionts. Furthermore, wild-type and T6SS-deficient S. marcescens strains achieved similar abundance and persistence in bee guts. Thus, an intact gut microbiota offers robust protection against this common pathogen, whose T6SSs do not confer the ability to compete with commensal species. IMPORTANCE Bacteria living within guts of animals can provide protection against infection by pathogens. Some pathogens have been shown to use a molecular weapon known as a T6SS to kill beneficial bacteria during invasion of the mouse gut. In this study, we examined how bacteria native to the honey bee gut work together to exclude the opportunistic pathogen Serratia marcescens. Although S. marcescens has a T6SS that can kill bacteria, bee gut bacteria seem resistant to its effects. This limitation may partially explain why ingestion of S. marcescens is rarely lethal to insects with healthy gut communities.


Asunto(s)
Antibiosis/fisiología , Abejas/microbiología , Microbioma Gastrointestinal/fisiología , Serratia marcescens/crecimiento & desarrollo , Animales , Fenómenos Fisiológicos Bacterianos , Abejas/inmunología , Simbiosis/fisiología , Sistemas de Secreción Tipo VI/fisiología
4.
Eur J Clin Microbiol Infect Dis ; 40(12): 2593-2596, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34363530

RESUMEN

A comparative analysis of the performance of the new selective chromogenic CHROMagar™-Serratia culture medium for detection and isolation of Serratia marcescens was undertaken. A total of 134 clinical isolates (95 S. marcescens with and without carbapenemase production and 39 non-S. marcescens isolates) and 96 epidemiological samples (46 rectal swabs and 50 from environmental surfaces) were studied. Diagnostic values when compared with CHROMagar™-Orientation medium were 96.8% sensitivity, 100% specificity, 100% positive predictive value and 88.5% negative predictive value. In conclusion, CHROMagar™-Serratia shows an excellent ability for differentiation of S. marcescens among clinical isolates and in environmental samples.


Asunto(s)
Técnicas Bacteriológicas/métodos , Medios de Cultivo/química , Infecciones por Serratia/microbiología , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/aislamiento & purificación , Agar/química , Agar/metabolismo , Técnicas Bacteriológicas/instrumentación , Compuestos Cromogénicos/química , Compuestos Cromogénicos/metabolismo , Medios de Cultivo/metabolismo , Humanos , Infecciones por Serratia/diagnóstico , Serratia marcescens/metabolismo
5.
Res Microbiol ; 172(6): 103869, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34333135

RESUMEN

Due to the emergence of multidrug-resistant bacteria, bacteriophages have become a viable alternative in controlling bacterial growth or biofilm formation. Biofilm is formed by extracellular polymeric substances (EPS) and is one of the factors responsible for increasing bacterial resistance. Bacteriophages have been studied as a bacterial control agent by use of phage enzymes or due to their bactericidal activities. A specific phage against Serratia marcescens was isolated in this work and was evaluated its biological and genomic aspects. The object of this study was UFV01, a bacteriophage belonging to the Podoviridae family, genus Teseptimavirus (group of lytic viruses), specific to the species S. marcescens, which may be related to several amino acid substitutions in the virus tail fibers. Despite this high specificity, the phage reduced the biofilm formation of several Escherichia coli strains without infecting them. UFV01 presents a relationship with phages of the genus Teseptimavirus, although it does not infect any of the E. coli strains evaluated, as these others do. All the characteristics make the phage an interesting alternative in biofilm control in hospital environments since small breaks in the biofilm matrix can lead to a complete collapse.


Asunto(s)
Biopelículas/crecimiento & desarrollo , Escherichia coli/crecimiento & desarrollo , Podoviridae/fisiología , Serratia liquefaciens/crecimiento & desarrollo , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/virología , Sustitución de Aminoácidos , Genoma Viral , Especificidad del Huésped , Concentración de Iones de Hidrógeno , Interacciones Microbianas , Podoviridae/clasificación , Podoviridae/genética , Podoviridae/aislamiento & purificación , Dominios Proteicos , Temperatura , Proteínas de la Cola de los Virus/química , Latencia del Virus
6.
J Basic Microbiol ; 61(6): 506-523, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33955034

RESUMEN

Prodigiosin is a natural red pigment derived primarily from secondary metabolites of microorganisms, especially Serratia marcescens. It can also be chemically synthesized. Prodigiosin has been proven to have antitumor, antibacterial, antimalaria, anti-insect, antialgae, and immunosuppressive activities, and is gaining increasing important in the global market because of its great potential application value in clinical medicine development, environmental treatment, preparation of food additives, and so on. Due to the low efficiency of prodigiosin chemical synthesis, high-level prodigiosin of production by microorganisms are necessary for prodigiosin applications. In this paper, the production of prodigiosin by microorganism in recent decades is reviewed. The methods and strategies for increasing the yield of prodigiosin are discussed from the aspects of medium composition, additives, factors affecting production conditions, strain modification, and fermentation methods.


Asunto(s)
Prodigiosina/biosíntesis , Vías Biosintéticas , Medios de Cultivo , Fermentación , Serratia marcescens/genética , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/metabolismo
7.
Arch Microbiol ; 203(5): 2269-2277, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-33638021

RESUMEN

Rhizopus species are opportunistic pathogens and cause infections which lead to deaths in individuals with the weakened immune system. Some strains of Rhizopus species have been detected to have a symbiotic relationship with bacteria. The toxicity of the Rhizopus species is important. Because strains harbouring endofungal bacteria are able to produce secondary metabolites and if endofungal bacteria are released from mycelium, serious problems can occur. We aimed to investigate the presence of endofungal bacteria in Rhizopus species isolated from food samples. Rhizopus species were isolated from different food samples. The presence of endofungal bacteria in the Rhizopus isolates was investigated. Rhizopus strains containing the endofungal bacteria were identified through phenotypic and genotypic methods. Universal primers amplifying bacterial 16S rRNA region were used to amplify 1.2-1.5-kb fragment from fungal metagenomic DNA. Sequence analysis of PCR products amplified from fungal metagenomic DNA was made. Fluorescence microscopy and scanning electron microscopy were used to visualize the presence of endofungal bacteria in fungal hyphae. According to our results, the Rhizopus strains is associated with Serratia marcescens, Pseudomonas fluorescens and Klebsiella pneumoniae. Until now there is no evidence that Pseudomonas fluorescens and Klebsiella pneumoniae were identified as endofungal. These species are opportunistic pathogen dangerous for humans. It is important for humans not only the presence of the fungi but also the presence of the endofungal bacteria in foods. Our work is important because it draws attention to the presence of endofungal bacteria in foods. Because there is danger releasing of a bacterium from the mycelium, it is likely to face sepsis or serious problems.


Asunto(s)
Hifa/fisiología , Klebsiella pneumoniae/aislamiento & purificación , Pseudomonas fluorescens/aislamiento & purificación , Rhizopus/metabolismo , Serratia marcescens/aislamiento & purificación , ADN de Hongos/genética , Microbiología de Alimentos , Humanos , Klebsiella pneumoniae/crecimiento & desarrollo , Micelio/química , Pseudomonas fluorescens/crecimiento & desarrollo , ARN Ribosómico 16S/genética , Rhizopus/genética , Serratia marcescens/crecimiento & desarrollo , Simbiosis
8.
Prep Biochem Biotechnol ; 51(7): 678-685, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33302794

RESUMEN

Microbial fermentation has become the main method to produce target compound. In this study, a 2-Keto-D-gluconic acid (2-KGA) producing mutant strain was obtained by mutation with rational screening methods. Meanwhile, prodigiosin was produced when the nitrogen source in the medium was changed to peptone and its fermentation conditions were evaluated to achieve high-efficient accumulation. The mutant strain SDSPY-136 was firstly identified as Serratia marcescens by morphological observation and 16S rDNA sequencing. The 2-KGA synthetic capacity of S. marcescens SDSPY-136 was evaluated by shake fermentation with 110 g/L glucose as substrates. For fermentation, 2-KGA yield, conversation rate and purity of SDSPY-136 reached 104.60 g/L, 95.10%, 99.11% in 72 h. The red pigment was extracted from the fermentation broth using acidic methanol and identified as prodigiosin by FT-IR. The optimal conditions were as follows: glycerol 20 g/L, peptone 20 g/L, MgSO415 g/L, pH 6.0, a 2% (v/v) inoculum, 30 °C and 200 rpm of shaking culture. Eventually, prodigiosin reached a yield of 9.89 g/Lafter shake culturing for 50 h under this condition. The mutant S. marcescens SDSPY-136 was shown to be promising for 2-KGA and prodigiosin production and a suitable object for prodigiosin metabolism research of S. marcescens.


Asunto(s)
Prodigiosina/biosíntesis , Serratia marcescens/crecimiento & desarrollo , Azúcares Ácidos/metabolismo , Mutación , Serratia marcescens/genética
9.
Biocontrol Sci ; 25(2): 73-80, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32507793

RESUMEN

Acanthamoeba is one of the organisms that cause corneal infection. In this study, attention was focused on potassium isostearate (iso-C18K, a branched chain fatty acid salt) for use in a multipurpose solution (MPS) against Acanthamoeba. An anti-amoebic test against Acanthamoeba castellanii ATCC 30010 (trophozoites type) was conducted. As a result, a growth reduction effect of 4 log units (99.99% suppression) was observed after incubation with 150 mM (5.0 w/v%) iso-C18K for 10 minutes. Furthermore, after the amoeba suspension was mixed with iso-C18K, disruption of cell membranes were observed, and the minimum amoebacidal concentration (MAC) at that time was 9.6 mM (0.31 w/v%). To evaluate the effectiveness as an MPS, assessment by verification tests was conducted using contact lenses. Reducing the concentration of iso-C18K caused a decrease in the number of viable cells, which was confirmed at a MAC of 1.2 mM (0.039 w/v%).


Asunto(s)
Acanthamoeba castellanii/efectos de los fármacos , Amebicidas/farmacología , Potasio/farmacología , Estearatos/farmacología , Trofozoítos/efectos de los fármacos , Acanthamoeba castellanii/crecimiento & desarrollo , Candida albicans/efectos de los fármacos , Candida albicans/crecimiento & desarrollo , Membrana Celular/efectos de los fármacos , Córnea , Fusarium/efectos de los fármacos , Fusarium/crecimiento & desarrollo , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/crecimiento & desarrollo , Serratia marcescens/efectos de los fármacos , Serratia marcescens/crecimiento & desarrollo , Soluciones , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Trofozoítos/crecimiento & desarrollo
10.
Molecules ; 25(12)2020 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-32545769

RESUMEN

The technology of microbial conversion provides a potential way to exploit compounds of biotechnological potential. The red pigment prodigiosin (PG) and other PG-like pigments from bacteria, majorly from Serratia marcescens, have been reported as bioactive secondary metabolites that can be used in the broad fields of agriculture, fine chemicals, and pharmacy. Increasing PG productivity by investigating the culture conditions especially the inexpensive carbon and nitrogen (C/N) sources has become an important factor for large-scale production. Investigations into the bioactivities and applications of PG and its related compounds have also been given increased attention. To save production cost, chitin and protein-containing fishery byproducts have recently been investigated as the sole C/N source for the production of PG and chitinolytic/proteolytic enzymes. This strategy provides an environmentally-friendly selection using inexpensive C/N sources to produce a high yield of PG together with chitinolytic and proteolytic enzymes by S. marcescens. The review article will provide effective references for production, bioactivity, and application of S. marcescens PG in various fields such as biocontrol agents and potential pharmaceutical drugs.


Asunto(s)
Quitina/química , Medios de Cultivo/química , Explotaciones Pesqueras , Prodigiosina/sangre , Serratia marcescens/crecimiento & desarrollo , Alimentos Marinos
11.
J Mater Chem B ; 8(12): 2454-2465, 2020 03 25.
Artículo en Inglés | MEDLINE | ID: mdl-32108210

RESUMEN

Tissue response to intestinal injury or disease releases pro-inflammatory host stress signals triggering microbial shift to pathogenic phenotypes. One such phenotype is increased protease production resulting in collagen degradation and activation of host matrix metalloproteinases contributing to tissue breakdown. We have shown that surgical injury depletes local intestinal phosphate concentration triggering bacterial virulence and that polyphosphate replenishment attenuates virulence and collagenolytic activity. Mechanistic studies of bacterial and host protease expression contributing to tissue breakdown are difficult to achieve in vivo necessitating the development of novel in vitro tissue models. Common techniques for screening in vitro protease activity, including gelatin zymography or fluorogenic protease-sensitive substrate kits, do not readily translate to 3D matrix degradation. Here, we report the application of an in vitro assay in which collagenolytic pathogens are cultured in the presence of a proteolytically degradable poly(ethylene) glycol scaffold and a non-degradable phosphate and/or polyphosphate nanocomposite hydrogel matrix. This in vitro platform enables quantification of pathogen-induced matrix degradation and screening of sustained release of phosphate-based therapeutic efficacy in attenuating protease expression. To evaluate matrix degradation as a function of bacterial enzyme levels secreted, we also present a novel method to quantify hydrogel degradation. This method involves staining protease-sensitive hydrogels with Sirius red dye to correlate absorbance of the degraded gel solution with hydrogel weight. This assay enables continuous monitoring and greater accuracy of hydrogel degradation kinetics compared to gravimetric measurements. Combined, the proposed in vitro platform and the presented degradation assay provide a novel strategy for screening efficacy of therapeutics in attenuating bacterial protease-induced matrix degradation.


Asunto(s)
Matriz Extracelular/metabolismo , Hidrogeles/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Péptido Hidrolasas/metabolismo , Fosfatos/metabolismo , Polietilenglicoles/metabolismo , Evaluación Preclínica de Medicamentos , Enterococcus faecalis/enzimología , Enterococcus faecalis/crecimiento & desarrollo , Humanos , Hidrogeles/química , Metaloproteinasa 9 de la Matriz/química , Metaloproteinasa 9 de la Matriz/aislamiento & purificación , Tamaño de la Partícula , Péptido Hidrolasas/química , Péptido Hidrolasas/aislamiento & purificación , Fosfatos/química , Polietilenglicoles/química , Pseudomonas aeruginosa/enzimología , Pseudomonas aeruginosa/crecimiento & desarrollo , Serratia marcescens/enzimología , Serratia marcescens/crecimiento & desarrollo , Propiedades de Superficie , Ingeniería de Tejidos
12.
Can J Microbiol ; 66(3): 243-255, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31922894

RESUMEN

Serratia marcescens is a facultatively anaerobic bacterium and the most recognized producer of the hydrophobic pigment prodigiosin. Previous work has shown that prodigiosin both increases ATP production during population lag phase and approximately doubles the stationary-phase cell yield. Here, we employed both batch and chemostat culture methods to investigate prodigiosin's role during high rate growth at low cell density as peak cellular ATP levels decline. Batch culture experiments utilizing artificial pigment induction showed an ATP reduction during low cell density growth. In addition, pigment induction during fixed growth rate chemostat culture revealed a negative correlation between cellular levels of prodigiosin and ATP (r = -0.95). Variable growth rate chemostat experiments showed an inverse relationship between ATP per cell and prodigiosin per cell during low-density growth but a direct relationship during high-density growth. Rate modeling of chemostat data quantified the pigment's effect on cellular levels of ATP for both population growth phases. Finally, prodigiosin production in a heterologous bacterium led to ATP decline. These data with intact cells complement the established in vitro proton import function of prodigiosin pigment and may indicate an energy-spilling function during high rate, low cell density growth.


Asunto(s)
Adenosina Trifosfato/metabolismo , Prodigiosina/biosíntesis , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/metabolismo , Filogenia , Serratia marcescens/genética
13.
Curr Protein Pept Sci ; 21(4): 357-368, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31429686

RESUMEN

Contact lens wear is generally safe and provides excellent vision. However, contact lens wear is often associated with the risk of developing ocular surface infection and inflammation, and in severe cases, the infection can result in loss of vision. Antimicrobial peptide-coated contact lenses have been made to help reduce the incidence of infection and inflammation. This paper reviews the research progress from conception, through the laboratory and preclinical tests to the latest information on clinical testing of an antimicrobial contact lens. We provide insights into the pathways followed and pitfalls that have been encountered. The journey has not always been linear or smooth, but has resulted in some of the first published clinical testing of antimicrobial peptide-coated contact lenses in humans. We hope this may help lead to the development and commercialisation of antimicrobial contact lenses in the future.


Asunto(s)
Antiinfecciosos/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Infecciones Bacterianas/prevención & control , Materiales Biocompatibles Revestidos/farmacología , Lentes de Contacto/microbiología , Queratitis/prevención & control , Micosis/prevención & control , Secuencia de Aminoácidos , Animales , Infecciones Bacterianas/etiología , Infecciones Bacterianas/microbiología , Infecciones Bacterianas/patología , Ensayos Clínicos como Asunto , Lentes de Contacto/efectos adversos , Evaluación Preclínica de Medicamentos , Humanos , Queratitis/etiología , Queratitis/microbiología , Queratitis/patología , Pruebas de Sensibilidad Microbiana , Micosis/etiología , Micosis/microbiología , Micosis/patología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/crecimiento & desarrollo , Pseudomonas aeruginosa/patogenicidad , Conejos , Serratia marcescens/efectos de los fármacos , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/patogenicidad , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus aureus/patogenicidad
14.
J Insect Physiol ; 118: 103939, 2019 10.
Artículo en Inglés | MEDLINE | ID: mdl-31493391

RESUMEN

Following pathogen attack in a host, widespread changes are induced in the host's gene expression, in particular those involved in the immune system, growth and survival. Epigenetic mechanisms have been suggested to be involved in the regulation of these changes through a number of mechanisms. DNA methylation is one of the important epigenetic processes that is carried out by DNA (cytosine-5) methyltransferase (DNMT) and alters expression of target genes. Here, we identified two putative sequences of DNMT (i.e. DNMT1 and DNMT2) from the transcriptome dataset of Helicoverpa armigera that showed high similarity to the homologous sequences in Bombyx mori. Domain architectures of DNMT1 and DNMT2 exhibit the unique pattern of DNMTs that highlights conserved function of these genes in different insects. To see if these genes play any role in bacterial infection, we challenged the fifth instar larvae of H. armigera by injecting Bacillus thuringiensis and Serratia marcescens cells into the hemolymph. Transcript levels of the DNMTs were analyzed by RT-qPCR. The results showed that the expression levels of DNMT1 and DNMT2 increased in the bacteria-injected larvae. Injection of the heat-killed bacteria also induced the expression of the DNMTs, but lower than that of the live bacteria. To determine whether these genes function during bacterial infection, we injected the inhibitor of DNMTs, 5-azacytidine (5-AZA), into the larvae and 24 h later, the bacterial cells were also injected into the larvae. Bacterial replication and larval mortality were analyzed in the treated and control insects. We found that 5-AZA reduced bacterial replication and also mortality of the bacterial-injected larvae regardless of the pathogenic bacterial species. Interestingly, the expression levels of antimicrobial peptides (AMPs) were also modulated following 5-AZA treatment. In conclusion, we showed that upregulation of the DNMTs in H. armigera following bacterial infections modulates AMPs and thereby affects the insect-bacteria interactions.


Asunto(s)
Péptidos Catiónicos Antimicrobianos/metabolismo , ADN (Citosina-5-)-Metiltransferasas/genética , Mariposas Nocturnas/genética , Mariposas Nocturnas/microbiología , Animales , Péptidos Catiónicos Antimicrobianos/genética , Azacitidina/farmacología , Bacillus thuringiensis/crecimiento & desarrollo , Metilación de ADN , Hemolinfa/microbiología , Interacciones Huésped-Patógeno/genética , Inmunidad Innata , Larva/enzimología , Larva/genética , Larva/microbiología , Mariposas Nocturnas/enzimología , Mariposas Nocturnas/inmunología , Serratia marcescens/crecimiento & desarrollo
15.
Rev Soc Bras Med Trop ; 52: e20180502, 2019 Jun 27.
Artículo en Inglés | MEDLINE | ID: mdl-31271619

RESUMEN

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.


Asunto(s)
Acinetobacter baumannii/efectos de los fármacos , Antibacterianos/farmacología , Bacterias Gramnegativas/efectos de los fármacos , Klebsiella pneumoniae/efectos de los fármacos , Aceites Volátiles/farmacología , Origanum/química , Serratia marcescens/efectos de los fármacos , Acinetobacter baumannii/crecimiento & desarrollo , Antibacterianos/clasificación , Proteínas Bacterianas , Carbapenémicos/farmacología , Farmacorresistencia Bacteriana Múltiple , Bacterias Gramnegativas/crecimiento & desarrollo , Humanos , Klebsiella pneumoniae/crecimiento & desarrollo , Pruebas de Sensibilidad Microbiana , Serratia marcescens/crecimiento & desarrollo , beta-Lactamasas
16.
PLoS One ; 14(5): e0216581, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31112570

RESUMEN

Adequate gut microbiota establishment is important for lifelong health. The aim was to sequentially analyze the gut microbiota establishment in low-birth-weight preterm neonates admitted to a single neonatal intensive care unit during their first 3 weeks of life, comparing two epidemiological scenarios. Seven control infants were recruited, and another 12 during a severe S. marcescens outbreak. Meconium and feces from days 7, 14, and 21 of life were collected. Gut microbiota composition was determined by 16S rDNA massive sequencing. Cultivable isolates were genotyped by pulsed-field gel electrophoresis, with four S. marcescens submitted for whole-genome sequencing. The expected bacterial ecosystem expansion after birth is delayed, possibly related to antibiotic exposure. The Proteobacteria phylum dominates, although with marked interindividual variability. The outbreak group considerably differed from the control group, with higher densities of Escherichia coli and Serratia to the detriment of Enterococcus and other Firmicutes. Curiously, obligate predators were only detected in meconium and at very low concentrations. Genotyping of cultivable bacteria demonstrated the high bacterial horizontal transmission rate that was confirmed with whole-genome sequencing for S. marcescens. Preterm infants admitted at NICU are initially colonized by homogeneous microbial communities, most of them from the nosocomial environment, which subsequently evolve according to the individual conditions. Our results demonstrate the hospital epidemiology pressure, particularly during outbreak situations, on the gut microbiota establishing process.


Asunto(s)
Infección Hospitalaria/microbiología , Brotes de Enfermedades , Heces/microbiología , Microbioma Gastrointestinal/genética , Recién Nacido de Bajo Peso/metabolismo , Recien Nacido Prematuro/metabolismo , Infecciones por Serratia/microbiología , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , ADN Ribosómico/genética , Femenino , Humanos , Lactante , Recién Nacido , Unidades de Cuidado Intensivo Neonatal , Masculino , Serratia marcescens/genética , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/aislamiento & purificación
17.
mBio ; 10(2)2019 04 02.
Artículo en Inglés | MEDLINE | ID: mdl-30940712

RESUMEN

Dysbiosis of the gut microbiome has been implicated in inflammatory bowel diseases. We have shown that levels of Candida tropicalis, along with those of Escherichia coli and Serratia marcescens, are significantly elevated in Crohn's disease (CD) patients. Here, we evaluated the ability of a novel probiotic to prevent and treat polymicrobial biofilms (PMB) formed by C. tropicalis with E. coli and S. marcescens Since Candida albicans has been reported to be elevated in CD patients, we investigated the interactions of C. albicans with these bacterial species in biofilm formation. We determined whether the interaction between Candida spp. and bacteria is specific by using Trichosporon inkin and Saccharomyces fibuligera as comparators. Additionally, the effects of probiotics on C. albicans germination and biofilm formation were determined. To determine the ability of the probiotic to prevent or treat mature biofilms, probiotic filtrate was added to the PMB at early (prevention) and mature (treatment) phases. Biofilm thickness and architecture were assessed by confocal scanning laser microscopy. The effects of the probiotic on germination were evaluated in the presence of serum. Exposure of C. tropicalis PMB to probiotic filtrate reduced biofilm matrix, decreased thickness, and inhibited hyphal formation. We showed that C. albicans or C. tropicalis formed significantly thicker PMB than control biofilms, indicating that this interaction is Candida specific. Treatment with probiotic filtrate inhibited C. albicans germination and prevented/treated C. albicans PMB. The designed probiotic may have utility in the management of biofilm-associated gastrointestinal diseases such as Crohn's and colorectal cancer.IMPORTANCE The effects of diversity of the gut microbiome on inflammation have centered mainly on bacterial flora. Recent research has implicated fungal species and their interactions with other organisms in the inflammatory process. New ways to restore microbial balance in the gut are being explored. Our goal was to identify beneficial probiotic strains that would antagonize these fungal and bacterial pathogens that are elevated in the inflamed gut, and which also have antibiofilm activity. Fungus-bacterium correlation analysis allowed us to identify candidate probiotic species that can antagonize microbial pathogens, which we subsequently incorporated into a novel probiotic formulation. Amylase, which is known to have some antibiofilm activity, was also added to the probiotic mixture. This novel probiotic may have utility for the management of inflammatory bowel diseases by disrupting polymicrobial biofilm formation.


Asunto(s)
Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Candida albicans/crecimiento & desarrollo , Escherichia coli/crecimiento & desarrollo , Probióticos/farmacología , Serratia marcescens/crecimiento & desarrollo , Antibiosis , Candida tropicalis/crecimiento & desarrollo , Interacciones Microbianas
18.
J Microbiol Biotechnol ; 29(4): 596-606, 2019 Apr 28.
Artículo en Inglés | MEDLINE | ID: mdl-30856708

RESUMEN

N-acyl-homoserine lactone quorum sensing (AHL-QS) has been shown to regulate many physiological behaviors in Serratia marcescens MG1. In the current study, the effects of AHL-QS on the biosynthesis of acid and neutral products by S. marcescens MG1 and its isogenic ∆swrI with or without supplementing exogenous N-hexanoyl-L-homoserine lactone (C6-HSL) were systematically investigated. The results showed that swrI disruption resulted in rapid pH drops from 7.0 to 4.8, which could be restored to wild type by supplementing C6-HSL. Furthermore, fermentation product analysis indicated that ∆swrI could lead to obvious accumulation for acidogenesis products such as lactic acid and succinic acid, especially excess acetic acid (2.27 g/l) produced at the early stage of fermentation, whereas solventogenesis products by ∆swrI appeared to noticeably decrease by an approximate 30% for acetoin during 32-48 h and by an approximate 20% for 2,3-butanediol during 24-40 h, when compared to those by wild type. Interestingly, the excess acetic acid produced could be removed in an AHL-QS-independent manner. Subsequently, quantitative real-time PCR was used to determine the mRNA expression levels of genes responsible for acidogenesis and solventogenesis and showed consistent results with those of product synthesis. Finally, by close examination of promoter regions of the analyzed genes, four putative luxI box-like motifs were found upstream of genes encoding acetyl-CoA synthase, lactate dehydrogenase, α-acetolactate decarboxylase, and Lys-like regulator. The information from this study provides a novel insight into the roles played by AHL-QS in switching from acidogenesis to solventogenesis in S. marcescens MG1.


Asunto(s)
Acil-Butirolactonas/farmacología , Metabolismo de los Hidratos de Carbono/efectos de los fármacos , Fermentación , Percepción de Quorum , Serratia marcescens/efectos de los fármacos , Serratia marcescens/metabolismo , Ácido Acético/metabolismo , Acetoína/metabolismo , Proteínas Bacterianas/genética , Biomasa , Butileno Glicoles/metabolismo , Metabolismo de los Hidratos de Carbono/genética , Carboxiliasas/genética , Perfilación de la Expresión Génica , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Genes Bacterianos/genética , Glucosa/metabolismo , Concentración de Iones de Hidrógeno , L-Lactato Deshidrogenasa/genética , Ácido Láctico/metabolismo , Redes y Vías Metabólicas/efectos de los fármacos , Redes y Vías Metabólicas/genética , ARN Mensajero/metabolismo , Alineación de Secuencia , Serratia marcescens/genética , Serratia marcescens/crecimiento & desarrollo , Ácido Succínico/metabolismo , Factores de Tiempo
19.
Microb Cell Fact ; 18(1): 2, 2019 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-30609918

RESUMEN

BACKGROUND: Biosurfactants are surface-active agents produced by microorganisms that have higher efficiency and stability, lower toxicity and higher biocompatibility and biodegradability than chemical surfactants. Despite its properties and potential application in a wide range of environmental and industrial processes, biosurfactants are still not cost-competitive when compared to their synthetic counterparts. Cost effective technologies and renewable raw substrates as agro-industrial and regional waste from northeast of Brazil as cassava flour wastewater, supplemented with lactose and corn oil are mainly the chemically media for growing microorganism and in turn the production of the biosurfactant of quality. This study aimed to obtained biosurfactant by Serratia marcescens UCP 1549 containing cassava flour wastewater (CWW), by application of a full-factorial design, as sustainable practices in puts the production process in promising formulation medium. The characterization of the biomolecule was carried out, as well as the determination of its stability and toxicity for cabbage seeds. In addition, its ability to stimulate seed germination for agriculture application and oil spill bioremediation were investigated. RESULTS: Serratia marcescens showed higher reduction of surface tension (25.92 mN/m) in the new medium containing 0.2% lactose, 6% cassava flour wastewater and 5% corn waste oil, after 72 h of fermentation at 28 °C and 150 rpm. The substrate cassava flour wastewater showed a promising source of nutrients for biosurfactant production. The isolate biosurfactant exhibited a CMC of 1.5% (w/v) and showed an anionic and polymeric structure, confirmed by infrared spectra. The biomolecule demonstrated high stability under different temperatures, salinity and pH values and non-toxicity against to cabbage seeds. Thus, exploring biosurfactant their potential role in seeds germinations and the promotion and agricultural applications was investigated. In addition, the effectiveness of biosurfactant for removal burned motor oil adsorbed in sand was verified. CONCLUSIONS: The use of medium containing CWW not only reduces the cost of process of biosurfactant production, but also the environmental pollution due to the inappropriate disposal of this residue. This fact, added to the high stability and non-toxicity of the biosurfactant produced by S. marcescens UCP 1549, confirms its high environmental compatibility, make it a sustainable biocompound that can be replace chemical surfactants in diverse industries. In addition, the effectiveness of biosurfactant for stimulate seed germination and removing burned motor oil from sand, suggests its suitability for agriculture and bioremediation applications.


Asunto(s)
Biodegradación Ambiental , Serratia marcescens/metabolismo , Tensoactivos/metabolismo , Reactores Biológicos , Brassica/efectos de los fármacos , Brassica/crecimiento & desarrollo , Germinación/efectos de los fármacos , Concentración de Iones de Hidrógeno , Semillas/efectos de los fármacos , Semillas/crecimiento & desarrollo , Serratia marcescens/química , Serratia marcescens/crecimiento & desarrollo , Cloruro de Sodio/química , Contaminantes del Suelo/química , Contaminantes del Suelo/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier , Tensión Superficial , Tensoactivos/química , Tensoactivos/toxicidad , Temperatura
20.
Microbiologyopen ; 8(6): e00761, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-30381897

RESUMEN

Studies by Alexander Gurwitsch in the 1920' s with onion root cells revealed the phenomenon of mitogenetic radiation. Subsequent works by Popp, Van Wijk, Quickenden, Tillbury, and Trushin have demonstrated a link between Gurwitsch's mitogenetic radiation and the biophoton, emissions of light correlated with biological processes. The present study seeks to expand upon these and other works to explore whether biophoton emissions of bacterial cultures is used as an information carrier of environmental stress. Bacterial cultures (Escherichia coli and Serratia marcescens) were incubated for 24 hr in 5 ml of nutrient broth to stationary phase and cell densities of ~107 cells/mL. Cultures of E. coli were placed upon a photomultiplier tube housed within a dark box. A second bacterial culture, either E. coli or S. marcescens, was placed in an identical dark box at a distance of 5 m and received injections of hydrogen peroxide. Spectral analyses revealed significant differences in peak frequencies of 7.2, 10.1, and 24.9 Hz in the amplitude modulation of the emitted biophoton signal with respect to whether a peroxide injection occurred or not, and whether the species receiving the injection was E. coli or S. marcescens. These and the subsequent results of discriminant functions suggest that bacteria may release biophotons as a non-local communication system in response to stress, and that these biophotons are species specific.


Asunto(s)
Escherichia coli/efectos de la radiación , Serratia marcescens/efectos de la radiación , Escherichia coli/crecimiento & desarrollo , Escherichia coli/fisiología , Luz , Serratia marcescens/crecimiento & desarrollo , Serratia marcescens/fisiología , Especificidad de la Especie
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