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1.
Environ Microbiol Rep ; 16(3): e13265, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38747207

RESUMO

Role of dust in Salmonella transmission on chicken farms is not well characterised. Salmonella Typhimurium (ST) infection of commercial layer chickens was investigated using a novel sprinkling method of chicken dust spiked with ST and the uptake compared to a conventional oral infection. While both inoculation methods resulted in colonisation of the intestines, the Salmonella load in liver samples was significantly higher at 7 dpi after exposing chicks to sprinkled dust compared to the oral infection group. Infection of chickens using the sprinkling method at a range of doses showed a threshold for colonisation of the gut and organs as low as 1000 CFU/g of dust. Caecal content microbiota analysis post-challenge showed that the profiles of chickens infected by the sprinkling and oral routes were not significantly different; however, both challenges induced differences when compared to the uninfected negative controls. Overall, the study showed that dust sprinkling was an effective way to experimentally colonise chickens with Salmonella and alter the gut microbiota than oral gavage at levels as low as 1000 CFU/g dust. This infection model mimics the field scenario of Salmonella infection in poultry sheds. The model can be used for future challenge studies for effective Salmonella control.


Assuntos
Galinhas , Poeira , Microbioma Gastrointestinal , Doenças das Aves Domésticas , Salmonelose Animal , Salmonella typhimurium , Animais , Galinhas/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Poeira/análise , Salmonelose Animal/microbiologia , Salmonelose Animal/prevenção & controle , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/prevenção & controle , Ceco/microbiologia , Fígado/microbiologia
2.
Science ; 382(6676): eadj3502, 2023 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-38096285

RESUMO

The human gut microbiome plays an important role in resisting colonization of the host by pathogens, but we lack the ability to predict which communities will be protective. We studied how human gut bacteria influence colonization of two major bacterial pathogens, both in vitro and in gnotobiotic mice. Whereas single species alone had negligible effects, colonization resistance greatly increased with community diversity. Moreover, this community-level resistance rested critically upon certain species being present. We explained these ecological patterns through the collective ability of resistant communities to consume nutrients that overlap with those used by the pathogen. Furthermore, we applied our findings to successfully predict communities that resist a novel target strain. Our work provides a reason why microbiome diversity is beneficial and suggests a route for the rational design of pathogen-resistant communities.


Assuntos
Microbioma Gastrointestinal , Interações Hospedeiro-Patógeno , Infecções por Klebsiella , Klebsiella pneumoniae , Infecções por Salmonella , Salmonella typhimurium , Animais , Humanos , Camundongos , Nutrientes/metabolismo , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/crescimento & desenvolvimento , Klebsiella pneumoniae/metabolismo , Salmonella typhimurium/genética , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/metabolismo , Simbiose , Vida Livre de Germes , Infecções por Klebsiella/microbiologia , Infecções por Salmonella/microbiologia , Escherichia coli/genética , Escherichia coli/metabolismo
3.
Nature ; 609(7925): 144-150, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35850148

RESUMO

Retrons are prokaryotic genetic retroelements encoding a reverse transcriptase that produces multi-copy single-stranded DNA1 (msDNA). Despite decades of research on the biosynthesis of msDNA2, the function and physiological roles of retrons have remained unknown. Here we show that Retron-Sen2 of Salmonella enterica serovar Typhimurium encodes an accessory toxin protein, STM14_4640, which we renamed as RcaT. RcaT is neutralized by the reverse transcriptase-msDNA antitoxin complex, and becomes active upon perturbation of msDNA biosynthesis. The reverse transcriptase is required for binding to RcaT, and the msDNA is required for the antitoxin activity. The highly prevalent RcaT-containing retron family constitutes a new type of tripartite DNA-containing toxin-antitoxin system. To understand the physiological roles of such toxin-antitoxin systems, we developed toxin activation-inhibition conjugation (TAC-TIC), a high-throughput reverse genetics approach that identifies the molecular triggers and blockers of toxin-antitoxin systems. By applying TAC-TIC to Retron-Sen2, we identified multiple trigger and blocker proteins of phage origin. We demonstrate that phage-related triggers directly modify the msDNA, thereby activating RcaT and inhibiting bacterial growth. By contrast, prophage proteins circumvent retrons by directly blocking RcaT. Consistently, retron toxin-antitoxin systems act as abortive infection anti-phage defence systems, in line with recent reports3,4. Thus, RcaT retrons are tripartite DNA-regulated toxin-antitoxin systems, which use the reverse transcriptase-msDNA complex both as an antitoxin and as a sensor of phage protein activities.


Assuntos
Antitoxinas , Bacteriófagos , Retroelementos , Salmonella typhimurium , Sistemas Toxina-Antitoxina , Antitoxinas/genética , Bacteriófagos/metabolismo , DNA Bacteriano/genética , DNA de Cadeia Simples/genética , Conformação de Ácido Nucleico , Prófagos/metabolismo , DNA Polimerase Dirigida por RNA/metabolismo , Retroelementos/genética , Salmonella typhimurium/genética , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/virologia , Sistemas Toxina-Antitoxina/genética
4.
Cell Rep ; 38(1): 110180, 2022 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-34986344

RESUMO

The gut microbiota benefits the host by limiting enteric pathogen expansion (colonization resistance), partially via the production of inhibitory metabolites. Propionate, a short-chain fatty acid produced by microbiota members, is proposed to mediate colonization resistance against Salmonella enterica serovar Typhimurium (S. Tm). Here, we show that S. Tm overcomes the inhibitory effects of propionate by using it as a carbon source for anaerobic respiration. We determine that propionate metabolism provides an inflammation-dependent colonization advantage to S. Tm during infection. Such benefit is abolished in the intestinal lumen of Salmonella-infected germ-free mice. Interestingly, S. Tm propionate-mediated intestinal expansion is restored when germ-free mice are monocolonized with Bacteroides thetaiotaomicron (B. theta), a prominent propionate producer in the gut, but not when mice are monocolonized with a propionate-production-deficient B. theta strain. Taken together, our results reveal a strategy used by S. Tm to mitigate colonization resistance by metabolizing microbiota-derived propionate.


Assuntos
Anaerobiose/fisiologia , Propionatos/metabolismo , Salmonelose Animal/patologia , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/metabolismo , Animais , Antibiose/fisiologia , Bacteroides thetaiotaomicron/genética , Bacteroides thetaiotaomicron/metabolismo , Feminino , Microbioma Gastrointestinal/fisiologia , Vida Livre de Germes , Intestinos/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Camundongos Knockout , Nitratos/metabolismo
5.
Meat Sci ; 184: 108697, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34687928

RESUMO

The aim of this research was to determine the effect of pectin coating made with essential oils and/or extracts of Thymus vulgaris (thyme) and Thymbra spicata (thymbra) on the preservation of aerobically packaged sliced bolognas during cold storage. The treatment made with essential oils resulted in a reduction of 1.73 log CFU/g of Salmonella typhimurium ATCC 14028. Also, pectin coating made with essential oil-treated sliced bolognas had the lowest total mesophilic bacteria (6.27 log CFU/g), and total lactic acid bacteria (1.72 CFU/g), in comparison to non-treated bolognas, with 7.65 log CFU/g for total mesophilic bacteria and 4.99 log CFU/g for lactic acid bacteria. Application of an emulsion significantly (P < 0.05) affected L*(lightness), a*(redness), and b*(yellowness) values. The essential oil treatment had the highest TBARS values at the end of the storage period. The pH was not affected by the treatment (P > 0.05), but storage had a significant (P < 0.05) effect on the pH values.


Assuntos
Filmes Comestíveis , Produtos da Carne/microbiologia , Óleos Voláteis , Pectinas , Cor , Microbiologia de Alimentos , Embalagem de Alimentos/métodos , Conservação de Alimentos/métodos , Lactobacillales/crescimento & desenvolvimento , Lamiaceae/química , Salmonella typhimurium/crescimento & desenvolvimento , Thymus (Planta)/química
6.
Microbiol Spectr ; 9(3): e0114221, 2021 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-34908471

RESUMO

Novel melanoidins are produced by the Maillard reaction. Here, melanoidins with high antibacterial activity were tested by examining various combinations of reducing sugars and amino acids as reaction substrates. Twenty-two types of melanoidins were examined by combining two reducing sugars (glucose and xylose) and eleven l-isomers of amino acids (alanine, arginine, glutamine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, and valine) to confirm the effects of these melanoidins on the growth of Listeria monocytogenes at 25°C. The melanoidins produced from the combination of d-xylose with either l-phenylalanine (Xyl-Phe) or l-proline (Xyl-Pro), for which absorbance at 420 nm was 3.5 ± 0.2, completely inhibited the growth of L. monocytogenes at 25°C for 48 h. Both of the melanoidins exhibited growth inhibition of L. monocytogenes which was equivalent to the effect of nisin (350 IU/mL). The antimicrobial spectrum of both melanoidins was also investigated for 10 different species of bacteria, including both Gram-positive and Gram-negative species. While Xyl-Phe-based melanoidin successfully inhibited the growth of Bacillus cereus and Brevibacillus brevis, Xyl-Pro-based melanoidin inhibited the growth of Salmonella enterica Typhimurium. However, no clear trend in the antimicrobial spectrum of the melanoidins against different bacterial species was observed. The findings in the present study suggest that melanoidins generated from xylose with phenylalanine and/or proline could be used as potential novel alternative food preservatives derived from food ingredients to control pathogenic bacteria. IMPORTANCE Although the antimicrobial effect of melanoidins has been reported in some foods, there have been few comprehensive investigations on the antimicrobial activity of combinations of reaction substrates of the Maillard reaction. The present study comprehensively investigated the potential of various combinations of reducing sugars and amino acids. Because the melanoidins examined in this study were produced simply by heating in an autoclave at 121°C for 60 min, the targeted melanoidins can be easily produced. The melanoidins produced from combinations of xylose with either phenylalanine or proline exhibited a wide spectrum of antibiotic effects against various pathogens, including Listeria monocytogenes, Bacillus cereus, and Salmonella enterica Typhimurium. Since the antibacterial effect of the melanoidins on L. monocytogenes was equivalent to that of a nisin solution (350 IU/mL), we might expect a practical application of melanoidins as novel food preservatives.


Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Conservantes de Alimentos/farmacologia , Polímeros/farmacologia , Aminoácidos/metabolismo , Bacillus cereus/efeitos dos fármacos , Bacillus cereus/crescimento & desenvolvimento , Bactérias/crescimento & desenvolvimento , Brevibacillus/efeitos dos fármacos , Brevibacillus/crescimento & desenvolvimento , Microbiologia de Alimentos/métodos , Glucose/metabolismo , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Reação de Maillard , Testes de Sensibilidade Microbiana , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento , Xilose/metabolismo
7.
Cell Mol Biol (Noisy-le-grand) ; 67(3): 24-34, 2021 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-34933736

RESUMO

The economic approaches for manufacturing the nanoparticles with physical and chemical effects and limited resistance to antibiotics have been progressed recently due to the rise of microbial resistance to antibiotics. This research aimed to study the antimicrobial efficacy of silver nanoparticles Ag, ZnO, and Tio2 nanoparticles against Salmonella typhimurium and Brucella abortus and Candida albicans. Two isolates of Salmonella and two isolates of Brucella abortus were isolated from food spastically meat and blood specimens, respectively. Candida albicans were isolated from the patient's mouth with oral candidiasis (oral thrush) and confirmed diagnosis by API 20C test. The antimicrobial susceptibility of Salmonella typhimurium and B. abortus isolates were performed against nine different antibiotics. Silver nanoparticles consisting of AgNPs size (90) nm, ZnO NPs size (20, 50) nm as well as TiO2 NPs size (10, 50) nm, were used. UV-Visible spectrophotometer was used to characterize silver nanoparticles. The highest resistance of Candida albicans was seen for fluconazole, Clotrimazole and Itraconazole. The results of the Minimum Inhibitory Concentration (MIC) of nanoparticles against Salmonella typhimurium showed the average MIC of Tio2-10nm and Tio2-50nm were 5000 and 2500 µg\ml for S1 and S2 isolates, respectively. The isolated Brucella abortus (B1 and B2) showed sensitivity to NPs with different MIC. The average MIC for Ag-90nm was 5000 and 2500 µg/ml for B1 and B2 isolates, respectively. The findings suggest NP solution has fungicidal and bactericidal impacts on the tested microorganisms so they can be suitable for multiple applications of the biomedical field such as developing new antimicrobial agents.


Assuntos
Bactérias/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Nanopartículas Metálicas/administração & dosagem , Prata/farmacologia , Titânio/farmacologia , Óxido de Zinco/farmacologia , Antibacterianos/administração & dosagem , Antibacterianos/química , Antibacterianos/farmacologia , Antifúngicos/administração & dosagem , Antifúngicos/química , Antifúngicos/farmacologia , Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Brucella abortus/efeitos dos fármacos , Brucella abortus/crescimento & desenvolvimento , Candida albicans/crescimento & desenvolvimento , Clotrimazol/administração & dosagem , Clotrimazol/química , Clotrimazol/farmacologia , Farmacorresistência Fúngica , Fluconazol/administração & dosagem , Fluconazol/química , Fluconazol/farmacologia , Humanos , Itraconazol/administração & dosagem , Itraconazol/química , Itraconazol/farmacologia , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana/métodos , Tamanho da Partícula , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento , Prata/administração & dosagem , Prata/química , Espectrofotometria/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Titânio/administração & dosagem , Titânio/química , Óxido de Zinco/administração & dosagem , Óxido de Zinco/química
8.
mBio ; 12(5): e0260821, 2021 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-34634938

RESUMO

For antibiotics with intracellular targets, effective treatment of bacterial infections requires the drug to accumulate to a high concentration inside cells. Bacteria produce a complex cell envelope and possess drug export efflux pumps to limit drug accumulation inside cells. Decreasing cell envelope permeability and increasing efflux pump activity can reduce intracellular accumulation of antibiotics and are commonly seen in antibiotic-resistant strains. Here, we show that the balance between influx and efflux differs depending on bacterial growth phase in Gram-negative bacteria. Accumulation of the fluorescent compound ethidium bromide (EtBr) was measured in Salmonella enterica serovar Typhimurium SL1344 (wild type) and efflux deficient (ΔacrB) strains during growth. In SL1344, EtBr accumulation remained low, regardless of growth phase, and did not correlate with acrAB transcription. EtBr accumulation in the ΔacrB strains was high in exponential phase but dropped sharply later in growth, with no significant difference from that in SL1344 in stationary phase. Low EtBr accumulation in stationary phase was not due to the upregulation of other efflux pumps but instead was due to decreased permeability of the envelope in stationary phase. Transcriptome sequencing (RNA-seq) identified changes in expression of several pathways that remodel the envelope in stationary phase, leading to lower permeability. IMPORTANCE This study shows that efflux is important for maintaining low intracellular accumulation only in actively growing cells and that envelope permeability is the predominant factor in stationary-phase cells. This conclusion means that (i) antibiotics with intracellular targets may be less effective in complex infections with nongrowing or slow-growing bacteria, where intracellular accumulation may be low; (ii) efflux inhibitors may be successful in potentiating the activity of existing antibiotics, but potentially only for bacterial infections where cells are actively growing; and (iii) the remodeling of the cell envelope prior to stationary phase could provide novel drug targets.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/genética , Salmonella typhimurium/metabolismo , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Transporte Biológico , Farmacorresistência Bacteriana Múltipla , Regulação Bacteriana da Expressão Gênica , Proteínas de Membrana Transportadoras/genética , Testes de Sensibilidade Microbiana , Salmonella typhimurium/efeitos dos fármacos
9.
Biochem Biophys Res Commun ; 582: 72-76, 2021 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-34695753

RESUMO

Type III secretion system (T3SS) plays a critical role in host cell invasion and pathogenesis of Salmonella. We recently identified the mycotoxin fusaric acid (FA) as a T3SS inhibitor of Salmonella. Herein, twenty-two diphenylsulfane derivatives were designed and synthesized using FA as a lead compound through scaffold hopping. Among them, SL-8 and SL-19 possessing strong anti-T3SS and anti-invasion activity were identified as T3SS inhibitors with improvement in potency as compared to FA. The inhibitory mechanisms on SPI-1 did not depend on the HilD-HilC-RtsA-HilA or PhoP-PhoQ pathway or the assembly of T3SS needle complex. Accordingly, we proposed that the inhibitory effects of SL-8 and SL-19 on SPI-1 probably influence the formation of SicA/InvF-effector complex or other related proteins.


Assuntos
Antibacterianos/síntese química , Proteínas de Bactérias/genética , Proteínas de Ligação a DNA/genética , Ácido Fusárico/análogos & derivados , Chaperonas Moleculares/genética , Salmonella typhimurium/efeitos dos fármacos , Fatores de Transcrição/genética , Sistemas de Secreção Tipo III/antagonistas & inibidores , Antibacterianos/farmacologia , Antígenos de Bactérias/genética , Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Produtos Biológicos , Células CACO-2 , Proteínas de Ligação a DNA/metabolismo , Flagelina/genética , Flagelina/metabolismo , Ácido Fusárico/farmacologia , Regulação Bacteriana da Expressão Gênica , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Chaperonas Moleculares/metabolismo , Salmonella typhimurium/genética , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/metabolismo , Transdução de Sinais , Relação Estrutura-Atividade , Fatores de Transcrição/metabolismo , Sistemas de Secreção Tipo III/genética , Sistemas de Secreção Tipo III/metabolismo
10.
Biomed Res Int ; 2021: 5604458, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34568492

RESUMO

Disease caused by antibiotic-resistant Salmonella is a serious clinical problem that poses a great threat to public health. The present study is aimed at assessing differences in bacterial kinetics with different antibiotic resistance profiles under environmental stress and at developing microbial tolerance models in lettuce during storage from 4 to 36°C. The drug-resistance phenotypes of 10 Salmonella Typhimurium (S. Typhimurium) isolates were examined using the broth microdilution method. The results of 10 S. Typhimurium isolates in the suspensions showed that a slow trend towards reduction of drug-sensitive (DS) isolates in relation to the others though without statistical difference. Compared to DS S. Typhimurium SA62, greater bacterial reduction was observed in multidrug-resistant (MDR) S. Typhimurium HZC3 during lettuce storage at 4°C (P < 0.05). It was likely that a cross-response between antibiotic resistance and food-associated stress tolerance. The greater growth in lettuce at 12°C was observed for DS S. Typhimurium SA62 compared to MDR S. Typhimurium HZC3 and was even statistically different (P < 0.05), while no significant difference was observed for bacterial growth between MDR S. Typhimurium HZC3 and DS S. Typhimurium SA62 strains in lettuce storage from 16 to 36°C (P > 0.05). The goodness-of-fit indices indicated the Log-linear primary model provided a satisfactory fit to describe the MDR S. Typhimurium HZC3 and DS S. Typhimurium SA62 survival at 4°C. A square root secondary model could be used to describe the effect of temperature (12, 16, 28, and 36°C) on the growth rates of S. Typhimurium HZC3 (adj - R 2 = 0.91, RMSE = 0.06) and S. Typhimurium SA62 (adj - R 2 = 0.99, RMSE = 0.01) derived from the Huang primary model. It was necessary to pay attention to the tolerance of antibiotic resistant bacteria under environmental stress, and the generated models could provide parts of the input data for microbial risk assessment of Salmonella with different antibiotic resistance profile in lettuce.


Assuntos
Antibacterianos/farmacologia , Resistência Microbiana a Medicamentos , Salmonella typhimurium/fisiologia , Estresse Fisiológico , Resistência Microbiana a Medicamentos/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Meio Ambiente , Lactuca/microbiologia , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Modelos Biológicos , Fenótipo , Reprodutibilidade dos Testes , Medição de Risco , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/isolamento & purificação , Estresse Fisiológico/efeitos dos fármacos
11.
Sci Rep ; 11(1): 18026, 2021 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-34504138

RESUMO

Salmonella Typhimurium is a human pathogen associated with eggs and egg-derived products. In Australia, it is recommended that eggs should be refrigerated to prevent condensation that can enhance bacterial penetration across the eggshell. Except for the United States, the guidelines on egg refrigeration are not prescriptive. In the current study, in-vitro and in-vivo experiments were conducted to understand the role of egg storage temperatures (refrigerated vs ambient) on bacterial load and the virulence genes expression of Salmonella Typhimurium. The in-vitro egg study showed that the load of Salmonella Typhimurium significantly increased in yolk and albumen stored at 25 °C. The gene expression study showed that ompR, misL, pefA, spvA, shdA, bapA, and csgB were significantly up-regulated in the egg yolk stored at 5 °C and 25 °C for 96 h; however, an in-vivo study revealed that mice infected with egg yolk stored at 25 °C, developed salmonellosis from day 3 post-infection (p.i.). Mice fed with inoculated egg yolk, albumen, or eggshell wash stored at refrigerated temperature did not show signs of salmonellosis during the period of the experiment. Data obtained in this study highlighted the importance of egg refrigeration in terms of improving product safety.


Assuntos
Ovos/microbiologia , Inocuidade dos Alimentos/métodos , Refrigeração/métodos , Intoxicação Alimentar por Salmonella/prevenção & controle , Infecções por Salmonella/prevenção & controle , Salmonella typhimurium/patogenicidade , Animais , Austrália , Galinhas , Contagem de Colônia Microbiana , Feminino , Microbiologia de Alimentos , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Intoxicação Alimentar por Salmonella/microbiologia , Intoxicação Alimentar por Salmonella/patologia , Infecções por Salmonella/microbiologia , Infecções por Salmonella/patologia , Salmonella typhimurium/genética , Salmonella typhimurium/crescimento & desenvolvimento , Temperatura , Virulência
12.
Sci Rep ; 11(1): 18173, 2021 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-34518559

RESUMO

Itaconic acid is produced by immune responsive gene 1 (IRG1)-coded enzyme in activated macrophages and known to play an important role in metabolism and immunity. In this study, mechanism of itaconic acid functioning as an anti-inflammatory metabolite was investigated with molecular biology and immunology techniques, by employing IRG1-null (prepared with CRISPR) and wild-type macrophages. Experimental results showed that itaconic acid significantly promoted the pentose phosphate pathway (PPP), which subsequently led to significantly higher NADPH oxidase activity and more reactive oxygen species (ROS) production. ROS production increased the expression of anti-inflammatory gene A20, which in turn decreased the production of inflammatory cytokines IL-6, IL-1ß and TNF-α. NF-κB, which can up-regulate A20, was also vital in controlling IRG1 and itaconic acid involved immune-modulatory responses in LPS-stimulated macrophage in this study. In addition, itaconic acid inhibited the growth of Salmonella typhimurium in cell through increasing ROS production from NADPH oxidase and the hatching of Schistosoma japonicum eggs in vitro. In short, this study revealed an alternative mechanism by which itaconic acid acts as an anti-inflammatory metabolite and confirmed the inhibition of bacterial pathogens with itaconic acid via ROS in cell. These findings provide the basic knowledge for future biological applications of itaconic acid in anti-inflammation and related pathogens control.


Assuntos
Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Via de Pentose Fosfato/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Succinatos/farmacologia , Animais , Citocinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/microbiologia , Macrófagos/patologia , Camundongos , NADPH Oxidases/metabolismo , NF-kappa B/metabolismo , Óvulo/efeitos dos fármacos , Células RAW 264.7 , Salmonella typhimurium/crescimento & desenvolvimento , Schistosoma japonicum/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos
13.
Cell Host Microbe ; 29(10): 1531-1544.e9, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34536347

RESUMO

The minimal genetic requirements for microbes to survive within multiorganism communities, including host-pathogen interactions, remain poorly understood. Here, we combined targeted gene mutagenesis with phenotype-guided genetic reassembly to identify a cooperative network of SPI-2 T3SS effector genes that are sufficient for Salmonella Typhimurium (STm) to cause disease in a natural host organism. Five SPI-2 effector genes support pathogen survival within the host cell cytoplasm by coordinating bacterial replication with Salmonella-containing vacuole (SCV) division. Unexpectedly, this minimal genetic repertoire does not support STm systemic infection of mice. In vivo screening revealed a second effector-gene network, encoded by the spv operon, that expands the life cycle of STm from growth in cells to deep-tissue colonization in a murine model of typhoid fever. Comparison between Salmonella infection models suggests how cooperation between effector genes drives tissue tropism in a pathogen group.


Assuntos
Proteínas de Bactérias/genética , Redes Reguladoras de Genes , Infecções por Salmonella/microbiologia , Salmonella typhimurium/genética , Animais , Proteínas de Bactérias/metabolismo , Citoplasma/microbiologia , Feminino , Ilhas Genômicas , Interações Hospedeiro-Patógeno , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Viabilidade Microbiana , Óperon , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/patogenicidade , Salmonella typhimurium/fisiologia , Tropismo , Sistemas de Secreção Tipo III/genética , Sistemas de Secreção Tipo III/metabolismo , Virulência
14.
Food Microbiol ; 100: 103853, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34416958

RESUMO

The combined effects of ethylenediaminetetraacetic acid (EDTA) and bacteriophage (phage) treatment of foodborne pathogens were investigated. Although viable counts for Campylobacter jejuni decreased by 1.5 log after incubation for 8 h in the presence of phage PC10, re-growth was observed thereafter. The combination of phage PC10 and 1 mM EDTA significantly inhibited the re-growth of C. jejuni. The viable counts for C. jejuni decreased by 2.6 log (P < 0.05) compared with that of the initial count after 24 h. Moreover, EDTA at 0.67 or 1.3 mM, combined with the specific lytic phages, also effectively inhibited the re-growth of phage-resistant cells of Campylobacter coli, Salmonella enterica serovar Enteritidis, and Salmonella enterica serovar Typhimurium. In addition, the combined effects of lytic phages and EDTA were investigated on the viability of Campylobacter in BHI broth at low temperatures followed by the optimum growth temperature. The re-growth of C. coli was significantly inhibited by the coexistence of 1.3 mM EDTA, and the viable counts of surviving bacteria was about the same as the initial viable count after the incubation. This is the first study demonstrating the combined use of lytic phages and EDTA is effective in inhibiting the re-growth of phage-resistant bacteria in Gram-negative bacteria.


Assuntos
Bacteriófagos/fisiologia , Campylobacter coli/crescimento & desenvolvimento , Campylobacter jejuni/crescimento & desenvolvimento , Ácido Edético/farmacologia , Salmonella enteritidis/crescimento & desenvolvimento , Salmonella typhimurium/crescimento & desenvolvimento , Campylobacter coli/efeitos dos fármacos , Campylobacter coli/virologia , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/virologia , Viabilidade Microbiana , Salmonella enteritidis/efeitos dos fármacos , Salmonella enteritidis/virologia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/virologia
15.
Food Microbiol ; 100: 103862, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34416962

RESUMO

Bacterial food poisoning cases due to Salmonella have been linked with a variety of poultry products. This study evaluated the effects of a Salmonella-specific Lytic bacteriophage and Lactobionic acid (LBA) on Salmonella Typhimurium DT 104 growth on raw chicken breast meat. Each chicken breast was randomly assigned to a treatment group (Control, DI water, phage 1%, phage 5%, LBA 10 mg/mL, LBA 20 mg/mL, and phage 5% + LBA 20 mg/mL) with four chicken breasts per group. Samples were inoculated with 106 CFU/mL of Salmonella and stored at 4 °C for 30 min. The inoculated chicken breasts were randomly assigned to different storage time (0 h, 1 h, 24 h, or 48 h). Both time and treatment showed significance reduction (P < 0.0001) of microbial growth. The weight loss was significantly different (P < 0.0001) between treatments. The LBA treatments were not effective when compared to the control group, but Lytic bacteriophage significantly reduced the amount of microbial growth.


Assuntos
Dissacarídeos/farmacologia , Conservação de Alimentos/métodos , Carne/microbiologia , Fagos de Salmonella/fisiologia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/virologia , Animais , Galinhas/microbiologia , Armazenamento de Alimentos , Salmonella typhimurium/crescimento & desenvolvimento
16.
Molecules ; 26(11)2021 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-34205065

RESUMO

Bacterial resistance to antibiotics due to increased efficiency of the efflux is a serious problem in clinics of infectious diseases. Knowledge of the factors affecting the activity of efflux pumps would help to find the solution. For this, fast and trustful methods for efflux analysis are needed. Here, we analyzed how the assay conditions affect the accumulation of efflux indicators ethidium (Et+) and tetraphenylphosphonium in Salmonella enterica ser. Typhimurium cells. An inhibitor phenylalanyl-arginyl-ß-naphtylamide was applied to evaluate the input of RND family pumps into the total efflux. In parallel to spectrofluorimetric analysis, we used an electrochemical assessment of Et+ concentration. The results of our experiments indicated that Et+ fluorescence increases immediately after the penetration of this indicator into the cells. However, when cells bind a high amount of Et+, the intensity of the fluorescence reaches the saturation level and stops reacting to the accumulated amount of this indicator. For this reason, electrochemical measurements provide more trustful information about the efficiency of efflux when cells accumulate high amounts of Et+. Measurements of Et+ interaction with the purified DNA demonstrated that the affinity of this lipophilic cation to DNA depends on the medium composition. The capacity of DNA to bind Et+ considerably decreases in the presence of Mg2+, Polymyxin B or when DNA is incubated in high ionic strength media.


Assuntos
DNA/química , Etídio/análise , Salmonella typhimurium/crescimento & desenvolvimento , Espermatozoides/química , Animais , Proteínas de Bactérias/metabolismo , Farmacorresistência Bacteriana Múltipla , Etídio/química , Masculino , Oniocompostos/química , Compostos Organofosforados/química , Salmão , Salmonella typhimurium/metabolismo , Espectrometria de Fluorescência , Espermatozoides/metabolismo
17.
mBio ; 12(3): e0109321, 2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34154399

RESUMO

Antimicrobial resistance (AMR) is a pressing global health crisis, which has been fueled by the sustained use of certain classes of antimicrobials, including fluoroquinolones. While the genetic mutations responsible for decreased fluoroquinolone (ciprofloxacin) susceptibility are known, the implications of ciprofloxacin exposure on bacterial growth, survival, and interactions with host cells are not well described. Aiming to understand the influence of inhibitory concentrations of ciprofloxacin in vitro, we subjected three clinical isolates of Salmonella enterica serovar Typhimurium to differing concentrations of ciprofloxacin, dependent on their MICs, and assessed the impact on bacterial growth, morphology, and transcription. We further investigated the differential morphology and transcription that occurred following ciprofloxacin exposure and measured the ability of ciprofloxacin-treated bacteria to invade and replicate in host cells. We found that ciprofloxacin-exposed S. Typhimurium is able to recover from inhibitory concentrations of ciprofloxacin and that the drug induces specific morphological and transcriptional signatures associated with the bacterial SOS response, DNA repair, and intracellular survival. In addition, ciprofloxacin-treated S. Typhimurium has increased capacity for intracellular replication in comparison to that of untreated organisms. These data suggest that S. Typhimurium undergoes an adaptive response under ciprofloxacin perturbation that promotes cellular survival, a consequence that may justify more measured use of ciprofloxacin for Salmonella infections. The combination of multiple experimental approaches provides new insights into the collateral effects that ciprofloxacin and other antimicrobials have on invasive bacterial pathogens. IMPORTANCE Antimicrobial resistance is a critical concern in global health. In particular, there is rising resistance to fluoroquinolones, such as ciprofloxacin, a first-line antimicrobial for many Gram-negative pathogens. We investigated the adaptive response of clinical isolates of Salmonella enterica serovar Typhimurium to ciprofloxacin, finding that the bacteria adapt in short timespans to high concentrations of ciprofloxacin in a way that promotes intracellular survival during early infection. Importantly, by studying three clinically relevant isolates, we were able to show that individual isolates respond differently to ciprofloxacin and that for each isolate, there was a heterogeneous response under ciprofloxacin treatment. The heterogeneity that arises from ciprofloxacin exposure may drive survival and proliferation of Salmonella during treatment and lead to drug resistance.


Assuntos
Antibacterianos/farmacologia , Ciprofloxacina/farmacologia , Viabilidade Microbiana/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Transcrição Gênica/efeitos dos fármacos , Proteínas de Bactérias/genética , Perfilação da Expressão Gênica , Humanos , Testes de Sensibilidade Microbiana , Infecções por Salmonella/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/fisiologia , Sorogrupo
18.
Int J Biol Macromol ; 184: 429-437, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-34166693

RESUMO

This study was carried out to characterize antioxidant activity, total phenolic content, and the phenolic and flavonoids profile of postbiotic of Pediococcus acidilactici and to evaluate the effects of postbiotics (10% and 50%) alone and in combination with chitosan coating (1%) on the microbial and chemical quality of chicken breast fillets during storage at 4 °C. Antioxidant activity and total phenolic content of the postbiotics were found to be 1291.02 ± 1.5 mg/L TEAC and 2336.11 ± 2.36 mg/L GAE, respectively. The most abundant phenolic was vanillic acid, followed by t-caffeic, gallic, and caftaric acids. The postbiotic-chitosan (50% + 1%) combination decreased L. monocytogenes and S. Typhimurium counts by 1.5 and 2.1 log10 CFU/g, respectively, compared to the control (P < 0.05). This combination decreased the total viable count (TVC), lactic acid bacteria (LAB), and psychrotrophic bacteria count compared to the control (P < 0.05). No differences were found in thiobarbituric acid (TBA) values among the samples during storage (P > 0.05). Postbiotic treatment did not significantly change the pH values and color properties of the breast fillets (P > 0.05). Postbiotic-chitosan combinations extended the shelf-life by up to 12 days compared to the control. In conclusion, the postbiotic-chitosan combination can be used to preserve and improve the microbial quality of chicken meat products.


Assuntos
Quitosana/farmacologia , Pediococcus acidilactici/química , Fenóis/farmacologia , Produtos Avícolas/análise , Animais , Produtos Biológicos/química , Produtos Biológicos/farmacologia , Galinhas , Quitosana/química , Sinergismo Farmacológico , Armazenamento de Alimentos , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Fenóis/química , Produtos Avícolas/microbiologia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento
19.
Food Microbiol ; 99: 103804, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34119097

RESUMO

The effectiveness of three novel "host defence peptides" identified in human Apolipoprotein B (ApoB) as novel antimicrobial and antibiofilm agents to be employed in food industry is reported. ApoB-derived peptides have been found to exert significant antimicrobial effects towards Salmonella typhimurium ATCC® 14028 and Salmonella enteritidis 706 RIVM strains. Furthermore, they have been found to retain antimicrobial activity under experimental conditions selected to simulate those occurring during food storage, transportation and heat treatment, and have been found to be endowed with antibiofilm properties. Based on these findings, to evaluate the applicability of ApoB-derived peptides as food biopreservatives, coating solutions composed by chitosan (CH) and an ApoB-derived peptide have been prepared and found to be able to prevent Salmonella cells attachment to different kinds of surfaces employed in food industry. Finally, obtained coating solution has been demonstrated to hinder microbial proliferation in chicken meat samples. Altogether, obtained findings indicate that ApoB-derived peptides are promising candidates as novel biopreservatives for food packaging.


Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Apolipoproteínas B/química , Conservantes de Alimentos/farmacologia , Animais , Antibacterianos/química , Peptídeos Catiônicos Antimicrobianos/química , Galinhas , Embalagem de Alimentos , Conservação de Alimentos , Conservantes de Alimentos/química , Armazenamento de Alimentos , Carne/microbiologia , Testes de Sensibilidade Microbiana , Salmonella enteritidis/efeitos dos fármacos , Salmonella enteritidis/crescimento & desenvolvimento , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento
20.
Food Microbiol ; 99: 103825, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34119110

RESUMO

This study aimed to investigate the effect of different growth temperatures on the resistance of Escherichia coli O157:H7 and Salmonella Typhimurium to low-energy X-ray irradiation. Irradiation of contaminated phosphate-buffered saline with 0.6 kGy X-ray decreased the counts of E. coli O157:H7 cultured at 37 °C to below the detection limit (<1.0 colony-forming unit (CFU)/mL) and those of E. coli O157:H7 cultured at 25 and 15 °C by 4.82 and 4.45 log CFU/mL, respectively. The viable counts of S. Typhimurium cultured at 37, 25, and 15 °C in phosphate-buffered saline decreased by 3.56, 3.08, and 2.75 log CFU/mL, respectively, after irradiation with 0.6 kGy X-ray. Irradiation of contaminated lettuce with 0.4 kGy decreased the counts of E. coli O157:H7 cultured at 37, 25, and 15 °C by 3.97, 3.45, and 3.10 log CFU/cm2, respectively, and those of S. Typhimurium by 4.41, 3.84, and 3.40 log CFU/cm2, respectively. Growth temperature influenced pathogen resistance to X-ray irradiation by modulating cellular membrane and DNA integrity, intracellular enzyme activity, and efflux pump function. The results of this study suggest that the stress resistance status of pathogenic bacteria cultured at different growth temperatures should be considered for the application of X-ray irradiation for fresh produce sterilization.


Assuntos
Escherichia coli O157/crescimento & desenvolvimento , Escherichia coli O157/efeitos da radiação , Lactuca/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/efeitos da radiação , Contagem de Colônia Microbiana , Contaminação de Alimentos/prevenção & controle , Irradiação de Alimentos , Folhas de Planta/microbiologia , Temperatura , Raios X
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