scholarly journals Simple rules govern the diversity of bacterial nicotianamine-like metallophores

2019 ◽  
Author(s):  
Clémentine Laffont ◽  
Catherine Brutesco ◽  
Christine Hajjar ◽  
Gregorio Cullia ◽  
Roberto Fanelli ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Pathogenic Bacteria ◽  
Gene Encoding ◽  
Nutritional Immunity ◽  
Simple Rules ◽  
Paenibacillus Mucilaginosus ◽  
Preferential Binding ◽  
Complete Reversal

ABSTRACTIn metal-scarce environments, some pathogenic bacteria produce opine-type metallophores mainly to face the host’s nutritional immunity. This is the case of staphylopine, pseudopaline and yersinopine, identified inStaphylococcus aureus,Pseudomonas aeruginosaandYersinia pestisrespectively. These metallophores are synthesized by two (CntLM) or three enzymes (CntKLM), CntM catalyzing the last step of biosynthesis using diverse substrates (pyruvate or α-ketoglutarate), pathway intermediates (xNA or yNA) and cofactors (NADH or NADPH), depending on the species. Here, we explored substrate specificity of CntM by combining bioinformatics and structural analysis with chemical synthesis and enzymatic studies. We found that NAD(P)H selectivity was mainly due to the amino acid at position 33 (S. aureusnumbering) which ensures a preferential binding to NADPH when it is an arginine. Moreover, whereas CntM fromP. aeruginosapreferentially uses yNA over xNA, the staphylococcal enzyme is not stereospecific. Most importantly, selectivity towards α-ketoacids is largely governed by a single residue at position 150 of CntM (S. aureusnumbering): an aspartate at this position ensures selectivity towards pyruvate whereas an alanine leads to the consumption of both pyruvate and α-ketoglutarate. Modifying this residue inP. aeruginosaled to a complete reversal of selectivity. Thus, opine-type metallophore diversity is mainly mediated by the absence/presence of acntKgene encoding a histidine racemase, and the presence of an aspartate/alanine at position 150 of CntM. These two simple rules predict the production of a fourth metallophore byPaenibacillus mucilaginosus, which was confirmedin vitroand called bacillopaline.

scholarly journals Simple rules govern the diversity of bacterial nicotianamine-like metallophores

2019 ◽  
Vol 476 (15) ◽  
pp. 2221-2233 ◽  
Author(s):  
Clémentine Laffont ◽  
Catherine Brutesco ◽  
Christine Hajjar ◽  
Gregorio Cullia ◽  
Roberto Fanelli ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Pathogenic Bacteria ◽  
Gene Encoding ◽  
Nutritional Immunity ◽  
Simple Rules ◽  
Paenibacillus Mucilaginosus ◽  
Preferential Binding ◽  
Complete Reversal

Abstract In metal-scarce environments, some pathogenic bacteria produce opine-type metallophores mainly to face the host's nutritional immunity. This is the case of staphylopine, pseudopaline and yersinopine, identified in Staphylococcus aureus, Pseudomonas aeruginosa and Yersinia pestis, respectively. Depending on the species, these metallophores are synthesized by two (CntLM) or three enzymes (CntKLM), CntM catalyzing the last step of biosynthesis using diverse substrates (pyruvate or α-ketoglutarate), pathway intermediates (xNA or yNA) and cofactors (NADH or NADPH). Here, we explored the substrate specificity of CntM by combining bioinformatic and structural analysis with chemical synthesis and enzymatic studies. We found that NAD(P)H selectivity is mainly due to the amino acid at position 33 (S. aureus numbering) which ensures a preferential binding to NADPH when it is an arginine. Moreover, whereas CntM from P. aeruginosa preferentially uses yNA over xNA, the staphylococcal enzyme is not stereospecific. Most importantly, selectivity toward α-ketoacids is largely governed by a single residue at position 150 of CntM (S. aureus numbering): an aspartate at this position ensures selectivity toward pyruvate, whereas an alanine leads to the consumption of both pyruvate and α-ketoglutarate. Modifying this residue in P. aeruginosa led to a complete reversal of selectivity. Thus, the diversity of opine-type metallophore is governed by the absence/presence of a cntK gene encoding a histidine racemase, and the amino acid residue at position 150 of CntM. These two simple rules predict the production of a fourth metallophore by Paenibacillus mucilaginosus, which was confirmed in vitro and called bacillopaline.

scholarly journals Staphylococcus aureus Serves as an Iron Source for Pseudomonas aeruginosa during In Vivo Coculture

2005 ◽  
Vol 187 (2) ◽  
pp. 554-566 ◽  
Author(s):  
Lauren M. Mashburn ◽  
Amy M. Jett ◽  
Darrin R. Akins ◽  
Marvin Whiteley
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Pathogenic Bacteria ◽  
Iron Acquisition ◽  
Low Oxygen ◽  
Dialysis Membrane ◽  
Opportunistic Human Pathogen ◽  
The Impact

ABSTRACT Pseudomonas aeruginosa is a gram-negative opportunistic human pathogen often infecting the lungs of individuals with the heritable disease cystic fibrosis and the peritoneum of individuals undergoing continuous ambulatory peritoneal dialysis. Often these infections are not caused by colonization with P. aeruginosa alone but instead by a consortium of pathogenic bacteria. Little is known about growth and persistence of P. aeruginosa in vivo, and less is known about the impact of coinfecting bacteria on P. aeruginosa pathogenesis and physiology. In this study, a rat dialysis membrane peritoneal model was used to evaluate the in vivo transcriptome of P. aeruginosa in monoculture and in coculture with Staphylococcus aureus. Monoculture results indicate that approximately 5% of all P. aeruginosa genes are differentially regulated during growth in vivo compared to in vitro controls. Included in this analysis are genes important for iron acquisition and growth in low-oxygen environments. The presence of S. aureus caused decreased transcription of P. aeruginosa iron-regulated genes during in vivo coculture, indicating that the presence of S. aureus increases usable iron for P. aeruginosa in this environment. We propose a model where P. aeruginosa lyses S. aureus and uses released iron for growth in low-iron environments.

Favorable effects in vitro and in vivo of two clinical isolates of Pseudomonas aeruginosa on nutritionally deficient Staphylococcus aureus strains

1973 ◽  
Vol 19 (8) ◽  
pp. 973-981 ◽  
Author(s):  
T. Gadbois ◽  
J. De Repentigny ◽  
L. G. Mathieu
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Peritoneal Cavity ◽  
Pathogenic Bacteria ◽  
Bacterial Species ◽  
Mixed Cultures ◽  
Rabbit Skin ◽  
Metabolic Activities

We have studied aspects of interbacterial ecology with nutritionally dependent Staphylococcus aureus strains; they were grown in association with Pseudomonas aeruginosa in systems of mixed cultures and infections in vitro in a semisynthetic medium and in vivo in mouse peritoneal cavity and rabbit skin. In mixed cultures and in P. aeruginosa culture filtrates, thymine and tryptophan deficiencies in staphylococci were partly overcome. This is probably because P. aeruginosa supplied the essential metabolites required to ensure growth; however, other metabolic activities could also be involved. Other experiments showed that the sensitivity of thymineless staphylococci to nucleoside inhibitions was alleviated. In mixed infections with P. aeruginosa, the S. aureus thymineless strain has shown a greater ability to survive in the peritoneal cavity of mice than when injected alone, even when one species was injected after the other with different doses of bacteria. The examination of the liquid from the peritoneal cavity of infected mice by fluorescence microscopy after fluorochroming with acridine orange or auramine O has revealed that Pseudomonas endotoxin seems to damage leucocytes and consequently reduces the phagocytosis of Staphylococcus cells.Necrosis in rabbit skin was mainly due to S. aureus when both species were injected together intradermally; the thymineless strain was less harmful than the parent strain.It seems that survival and even growth of nutritionally dependent strains of a bacterial species can be favored by the metabolic activities of another species in mixed cultures and infections, in this instance S. aureus by P. aeruginosa. This phenomenon among others could be a determinant of bacterial pathogenicity for nutritionally dependent pathogenic bacteria; thus associated organisms could determine the effective pathogenicity of nutritionally dependent bacteria by contributing essential nutrilites at the site where infection is initiated.

scholarly journals Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth

2022 ◽  
Vol 82 ◽  
Author(s):  
I. D. C. Galo ◽  
R. P. Prado ◽  
W. G. Dos Santos
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Visible Light ◽  
Blue Light ◽  
Bacterial Infections ◽  
Pathogenic Bacteria ◽  
Bacterial Species ◽  
Red Light ◽  
High Rate

Abstract The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm2, 13.19 mW/cm2 and 603.44 J/cm2, 27.93 mW/cm2 respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.

scholarly journals Silver Sulfadiazine Eradicates Antibiotic-Tolerant Staphylococcus aureus and Pseudomonas aeruginosa Biofilms in Patients with Infected Diabetic Foot Ulcers

2020 ◽  
Vol 9 (12) ◽  
pp. 3807
Author(s):  
Enea Gino Di Domenico ◽  
Barbara De Angelis ◽  
Ilaria Cavallo ◽  
Francesca Sivori ◽  
Fabrizio Orlandi ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Diabetic Foot ◽  
Pathogenic Bacteria ◽  
Topical Administration ◽  
Silver Sulfadiazine ◽  
Foot Ulcers ◽  
Diabetic Foot Ulcers ◽  
Vitro Analysis

Infections are among the most frequent and challenging events in diabetic foot ulcers (DFUs). Pathogenic bacteria growing in biofilms within host tissue are highly tolerant to environmental and chemical agents, including antibiotics. The present study was aimed at assessing the use of silver sulfadiazine (SSD) for wound healing and infection control in 16 patients with DFUs harboring biofilm-growing Staphylococcus aureus and Pseudomonas aeruginosa. All patients received a treatment based on a dressing protocol including disinfection, cleansing, application of SSD, and application of nonadherent gauze, followed by sterile gauze and tibio-breech bandage, in preparation for toilet surgery after 30 days of treatment. Clinical parameters were analyzed by the T.I.M.E. classification system. In addition, the activity of SSD against biofilm-growing S. aureus and P. aeruginosa isolates was assessed in vitro. A total of 16 patients with S. aureus and P. aeruginosa infected DFUs were included in the study. Clinical data showed a statistically significant (p < 0.002) improvement of patients’ DFUs after 30 days of treatment with SSD with significant amelioration of all the parameters analyzed. Notably, after 30 days of treatment, resolution of infection was observed in all DFUs. In vitro analysis showed that both S. aureus and P. aeruginosa isolates developed complex and highly structured biofilms. Antibiotic susceptibility profiles indicated that biofilm cultures were significantly (p ≤ 0.002) more tolerant to all tested antimicrobials than their planktonic counterparts. However, SSD was found to be effective against fully developed biofilms of both S. aureus and P. aeruginosa at concentrations below those normally used in clinical preparations (10 mg/mL). These results strongly suggest that the topical administration of SSD may represent an effective alternative to conventional antibiotics for the successful treatment of DFUs infected by biofilm-growing S. aureus and P. aeruginosa.

ATIVIDADE ANTIMICROBIANA IN VITRO DO EXTRATO AQUOSO, HIDROALCOÓLICO E ALCOÓLICO DE FOLHAS DE ESPÉCIES DA FAMÍLIA LAMIACEAE

2018 ◽  
Vol 4 ◽  
Author(s):  
Karlynne Freire Mendonça ◽  
José Klauber Roger Carneiro ◽  
Maria Auxiliadora Silva Oliveira
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Enterococcus Faecalis ◽  
Streptococcus Pyogenes ◽  
Mentha Arvensis ◽  
Ocimum Gratissimum ◽  
Mueller Hinton ◽  
Plectranthus Amboinicus

Objetivos: avaliar a atividade antimicrobiana em extrato aquoso, hidroalcoólico e alcoólico das folhas de espécies da família Lamiaceae frente a bactérias de interesse. Método: Foram escolhidas quatro espécies: Ocimum gratissimum, Plectranthus amboinicus, Mentha arvensis e Plectranthus barbatus. A partir das folhas foram confeccionados os extratos aquoso, hidroalcoólico e alcoólico nas concentrações 100mg/mL, 50mg/mL e 25mg/mL. Foram selecionadas as bactérias Streptococcus pyogenes, Enterococcus faecalis, Staphylococcus aureus e Pseudomonas aeruginosa para os ensaios de antibiose em Ágar Mueller-Hinton. Resultados: P. barbatus, em seu extrato hidroalcoólico mostrou ativo nas três concentrações para bactéria S. aureus, e ainda foi ativo para P. aeruginosa, demonstrando no extrato alcoólico atividade frente as bactérias. Para M. arvensis e P. amboinicus, seus extratos hidroalcoólico e alcoólico apresentaram atividade para S. aureus. Conclusão: Sugere-se que as espécies em questão apresentem boa atividade antimicrobiana, sendo necessária a realização de mais estudos para melhor entender esse mecanismo.

Activité antibactérienne de l’huile essentielle de Pelargonium x asperum et son potentiel synergique avec la nisine

2019 ◽  
Vol 17 (3) ◽  
pp. 140-148 ◽  
Author(s):  
A. Ouelhadj ◽  
L. Ait Salem ◽  
D. Djenane
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Nous Avons ◽  
Activité Antibactérienne

Ce travail vise l’étude de l’activité antibactérienne de l’huile essentielle (HE) de Pelargoniumx asperum et de la bactériocine, la nisine seul et en combinaison vis-à-vis de six bactéries dont quatre sont multirésistantes d’origine clinique. L’activité antibactérienne in vitro a été évaluée par la méthode de diffusion sur gélose. La concentration minimale inhibitrice (CMI) est aussi déterminée pour HE. Les résultats ont révélé une activité antibactérienne significative exercée par HE visà-vis de Staphylococcus aureus (ATCC 43300), Staphylococcus aureus et Escherichia coli avec des diamètres d’inhibition de 36,00 ; 22,50 et 40,00 mm, respectivement. Cependant, l’HE de Pelargonium asperum a montré une activité antibactérienne supérieure par rapport à la nisine. Les valeurs des CMI rapportées dans cette étude sont comprises entre 1,98–3,96 μl/ml. Les combinaisons réalisées entre HE et la nisine ont montré un effet additif vis-à-vis de Escherichia coli (ATCC 25922) avec (50 % HE Pelargonium asperum + 50 % nisine). Par contre, nous avons enregistré une synergie vis-à-vis de Klebsiella pneumoniae avec (75 % HE Pelargonium asperum + 25 % nisine) et contre Pseudomonas aeruginosa avec les trois combinaisons testées. Les résultats obtenus permettent de dire que l’HE de Pelargonium asperum possède une activité antibactérienne ainsi que sa combinaison avec la nisine pourrait représenter une bonne alternative pour la lutte contre l’antibiorésistance.

Exploración de la actividad antibacteriana de Metformina frente a Escherichia coli, Staphylococcus aureus y Pseudomonas aeruginosa

Bionatura ◽  
2020 ◽  
Vol 5 (4) ◽  
pp. 1335-1339
Author(s):  
Pool Marcos-Carbajal ◽  
Christian Allca-Muñoz ◽  
Ángel Urbano-Niño ◽  
Alberto Salazar-Granara
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa

El objetivo del estudio es determinar la actividad antibacteriana de Metformina frente a Escherichia coli, Staphylococcus aureus y Pseudomonas aeruginosa. Se evaluó la actividad antibacteriana mediante la técnica de Kirby Bauer. Se utilizó cepas de Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923) y Pseudomonas aeruginosa (ATCC 27853), las cuales se expusieron a Metformina en concentraciones de 250 mg y 500 mg, Ciprofloxacino (CIP) 5 µg, Imipenem (IPM) 10 µg, y Cefoxitin (FOX) 30 µg. Frente a Escherichia coli, Staphylococcus aureus y Pseudomonas aeruginosa se presentó un halo de inhibición de 6 mm. para Metformina 250 mg, 6 mm. para Metformina 500 mg, y un halo de inhibición >25 mm. con el uso de Ciprofloxacino 5 µg, Cefoxitin 30 µg, e Imipenem 10 µg respectivamente. En conclusion, In vitro Metformina a dosis de 250 y 500 mg, no presentó efecto antibacteriano frente a Escherichia coli, Staphylococcus aureus y Pseudomonas aeruginosa.

scholarly journals Exogenous Alginate Protects Staphylococcus aureus from Killing by Pseudomonas aeruginosa

2019 ◽  
Vol 202 (8) ◽  
Author(s):  
Courtney E. Price ◽  
Dustin G. Brown ◽  
Dominique H. Limoli ◽  
Vanessa V. Phelan ◽  
George A. O’Toole
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Physical Space ◽  
Late Time ◽  
Protective Effects ◽  
Content Type ◽  
Alginate Production ◽  
The Impact

ABSTRACT Cystic fibrosis (CF) patients chronically infected with both Pseudomonas aeruginosa and Staphylococcus aureus have worse health outcomes than patients who are monoinfected with either P. aeruginosa or S. aureus. We showed previously that mucoid strains of P. aeruginosa can coexist with S. aureus in vitro due to the transcriptional downregulation of several toxic exoproducts typically produced by P. aeruginosa, including siderophores, rhamnolipids, and HQNO (2-heptyl-4-hydroxyquinoline N-oxide). Here, we demonstrate that exogenous alginate protects S. aureus from P. aeruginosa in both planktonic and biofilm coculture models under a variety of nutritional conditions. S. aureus protection in the presence of exogenous alginate is due to the transcriptional downregulation of pvdA, a gene required for the production of the iron-scavenging siderophore pyoverdine as well as the downregulation of the PQS (Pseudomonas quinolone signal) (2-heptyl-3,4-dihydroxyquinoline) quorum sensing system. The impact of exogenous alginate is independent of endogenous alginate production. We further demonstrate that coculture of mucoid P. aeruginosa with nonmucoid P. aeruginosa strains can mitigate the killing of S. aureus by the nonmucoid strain of P. aeruginosa, indicating that the mechanism that we describe here may function in vivo in the context of mixed infections. Finally, we investigated a panel of mucoid clinical isolates that retain the ability to kill S. aureus at late time points and show that each strain has a unique expression profile, indicating that mucoid isolates can overcome the S. aureus-protective effects of mucoidy in a strain-specific manner. IMPORTANCE CF patients are chronically infected by polymicrobial communities. The two dominant bacterial pathogens that infect the lungs of CF patients are P. aeruginosa and S. aureus, with ∼30% of patients coinfected by both species. Such coinfected individuals have worse outcomes than monoinfected patients, and both species persist within the same physical space. A variety of host and environmental factors have been demonstrated to promote P. aeruginosa-S. aureus coexistence, despite evidence that P. aeruginosa kills S. aureus when these organisms are cocultured in vitro. Thus, a better understanding of P. aeruginosa-S. aureus interactions, particularly mechanisms by which these microorganisms are able to coexist in proximal physical space, will lead to better-informed treatments for chronic polymicrobial infections.

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