RESUMEN

Alternative strategies for controlling Staphylococcus aureus and other pathogens have been continuously investigated, with nisin, a bacteriocin widely used in the food industry as a biopreservative, gaining increasing attention. In addition to its antimicrobial properties, bacteriocins have significant effects on genome functionality even at inhibitory concentrations. This study investigated the impact of subinhibitory concentrations of nisin on S. aureus. Culturing in the presence of 0.625 µmol l-1 nisin, led to the increased relative expression of hla, saeR, and sarA, genes associated with virulence while expression of the sea gene, encoding staphylococcal enterotoxin A (SEA), decreased. In an in vivo experiment, Galleria mellonella larvae inoculated with S. aureus cultured in the presence of nisin exhibited 97% mortality at 72 h post-infection, compared to over 40% of larvae mortality in larvae infected with S. aureus. A comprehensive understanding of the effect of nisin on the transcriptional response of virulence genes and the impact of these changes on the virulence of S. aureus can contribute to assessing the application of this bacteriocin in food and medical contexts.

Asunto(s)

Antibacterianos , Larva , Mariposas Nocturnas , Nisina , Staphylococcus aureus , Nisina/farmacología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidad , Animales , Virulencia/genética , Larva/microbiología , Larva/efectos de los fármacos , Antibacterianos/farmacología , Mariposas Nocturnas/microbiología , Infecciones Estafilocócicas/microbiología , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Factores de Virulencia/genética , Pruebas de Sensibilidad Microbiana

RESUMEN

IMPORTANCE: Bovine mastitis, predominantly associated with gram-positive Staphylococcus aureus, poses a significant threat to dairy cows, leading to a decline in milk quality and volume with substantial economic implications. OBJECTIVE: This study investigated the incidence, virulence, and antibiotic resistance of S. aureus associated with mastitis in dairy cows. METHODS: Fifty milk-productive cows underwent a subclinical mastitis diagnosis, and the S. aureus strains were isolated. Genomic DNA extraction, sequencing, and bioinformatic analysis were performed, supplemented by including 124 S. aureus genomes from cows with subclinical mastitis to enhance the overall analysis. RESULTS: The results revealed a 42% prevalence of subclinical mastitis among the cows tested. Genomic analysis identified 26 sequence types (STs) for all isolates, with Mexican STs belonging primarily to CC1 and CC97. The analyzed genomes exhibited multidrug resistance to phenicol, fluoroquinolone, tetracycline, and cephalosporine, which are commonly used as the first line of treatment. Furthermore, a similar genomic virulence repertoire was observed across the genomes, encompassing the genes related to invasion, survival, pathogenesis, and iron uptake. In particular, the toxic shock syndrome toxin (tss-1) was found predominantly in the genomes isolated in this study, posing potential health risks, particularly in children. CONCLUSION AND RELEVANCE: These findings underscore the broad capacity for antibiotic resistance and pathogenicity by S. aureus, compromising the integrity of milk and dairy products. The study emphasizes the need to evaluate the effectiveness of antibiotics in combating S. aureus infections.

Asunto(s)

Genoma Bacteriano , Mastitis Bovina , Infecciones Estafilocócicas , Staphylococcus aureus , Animales , Bovinos , Mastitis Bovina/microbiología , México/epidemiología , Femenino , Staphylococcus aureus/genética , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/patogenicidad , Infecciones Estafilocócicas/veterinaria , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/epidemiología , Virulencia/genética , Antibacterianos/farmacología , Farmacorresistencia Bacteriana/genética

RESUMEN

BACKGROUND: The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) has changed in recent years. The present article is intended to establish differences between clinical, laboratory and imaging findings and outcomes of MSSA and MRSA infections, as well as among subgroups of infection such as skin and soft tissue infection, osteoarticular, bacteremia or pneumonia in a pediatric population from Bogota, Colombia. METHODS: Retrospective cohort study using clinical records of patients under 18 years of age treated at the participating centers in Bogota, Colombia, between 2014 and 2018. The first positive S. aureus culture was studied. MSSA and MRSA were compared. The χ2 test, Fisher exact test, and Kruskal-Wallis test were calculated, and the statistical significance was presented using the difference and its 95% CI. RESULTS: Five hundred fifty-one patients were included; 211 (38%) corresponded to MRSA and 340 (62%) to MSSA for a total of 703 cultures. A significantly higher probability of having an MSSA infection than MRSA was found in patients with previous heart disease (3.3% vs. 0.5%), neurologic disease (5.9% vs. 2.5%), recent major surgeries (11% vs. 5%) or who has an implanted device (11% vs. 4%). In contrast, in severe MRSA infections (bacteremia, osteoarticular infections and pneumonia), a higher rate of complications was seen (admission to the pediatric intensive care unit, mechanical ventilation and vasoactive support), and in osteoarticular MRSA, more than 1 surgery per case was seen (89% vs. 61%). Laboratory results and mortality were similar. CONCLUSIONS: MRSA was associated with a more severe course in bacteremia, osteoarticular infections and pneumonia. Some classical risk factors associated with MRSA infections were found to be related to MSSA. In general, with the exception of skin and soft tissue infection, there was an increased risk of pediatric intensive care unit admission and mechanical and inotropic support with MRSA in a pediatric population.

Asunto(s)

Staphylococcus aureus Resistente a Meticilina/patogenicidad , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/patogenicidad , Adolescente , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Niño , Preescolar , Estudios de Cohortes , Colombia/epidemiología , Femenino , Hospitalización , Humanos , Lactante , Masculino , Meticilina/farmacología , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Estudios Retrospectivos , Factores de Riesgo , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus/efectos de los fármacos

RESUMEN

Biophysical characterization of antimicrobial peptides helps to understand the mechanistic aspects of their action. The physical behavior of the KR-12 antimicrobial peptide (e.g. orientation and changes in secondary structure), was analyzed after interactions with a Staphylococcus aureus membrane model and solid surfaces. We performed antimicrobial tests using Gram-positive S. aureus (ATCC 25923) bacteria. Moreover, Langmuir-Blodgett experiments showed that the synthetic peptide can disturb the lipidic membrane at a concentration lower than the Minimum Inhibitory Concentration, thus confirming that KR-12/lipid interactions are involved. Partially- and fully-deactivated KR-12 hybrid samples were obtained by physisorption and covalent immobilization in chitosan/silica and glyoxal-rich solid supports. The correlation of Langmuir-Blodgett data with the α-helix formation, followed by FTIR-ATR in a frozen-like state, and the antimicrobial activity showed the importance of these interactions and conformation changes on the first step action mode of this peptide. This is the first time that material science (immobilization in solid surfaces assisted by FTIR-ATR analysis in frozen-like state) and physical (Langmuir-Blodgett/Schaefer) approaches are combined for exploring mechanistic aspects of the primary action mode of the KR-12 antimicrobial peptide against S. aureus.

Asunto(s)

Antibacterianos/química , Péptidos Antimicrobianos/química , Catelicidinas/química , Lípidos/antagonistas & inhibidores , Fragmentos de Péptidos/química , Antibacterianos/farmacología , Catelicidinas/farmacología , Quitosano/química , Quitosano/farmacología , Humanos , Lípidos/química , Lípidos de la Membrana/antagonistas & inhibidores , Pruebas de Sensibilidad Microbiana , Fragmentos de Péptidos/farmacología , Espectroscopía Infrarroja por Transformada de Fourier , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/patogenicidad

RESUMEN

Introdução: Diariamente, estamos rodeados por microrganismos, e diversas situações favorecem essa aproximação. Nesse contexto, as cédulas de dinheiro se destacam como possível fonte de transmissão de patógenos, como enteroparasitas e bactérias, uma vez que são manuseadas por inúmeras pessoas. Objetivo e Método: Em vista disso, este estudo teve como objetivo elaborar um levantamento dos estudos realizados nos últimos 20 anos referentes à contaminação das cédulas de dinheiro por enteroparasitas e bactérias patogênicas. Resultados: Os resultados demonstraram que os enteroparasitas identificados com maior frequência nos estudos foram Ascaris lumbricoides, Entamoebacoli (não patogênico) e ancilostomídeos. Com relação à pesquisa de bactérias, Staphylococcus aureus, Klebsiellasp, Escherichia coli e Enterobactersp foram as mais detectadas. Esses dados evidenciam que existe a contaminação das cédulas de dinheiro por bactérias e enteroparasitas, confirmando seu papel como possível fonte de contaminação. Conclusão: Dessa forma, ressalta-se a importância da melhora nos hábitos de higiene básica como estratégia para limitar o ciclo desses patógenos.

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Introduction: Every day, we are surrounded by microorganisms, and several situations favor this approximation. In this context, money bills are a possible source for pathogens transmission, such as enteroparasites and bacteria, as they are handled by countless people. Objective and Method: Thus, we carried out a survey study considering the last 20 years of research related to money bills contamination by enteroparasites and pathogenic bacteria. Results: The results showed that the most frequently identified entheroparasites in the studies were Ascaris lumbricoides, Entamoeba coli (not pathogen), and hookworms. Regarding the bacteria research, the most frequently detected were Staphylococcus aureus, Klebsiella sp, Escherichia coli and Enterobacter sp. Conclusion: Data shows that money bills contamination by bacteria and enteroparasites exists, confirming its role as a contam-ination source. Thereby, the importance of better basic hygiene habits as a strategy to limit the pathogen's cycle is reinforced.

Asunto(s)

Indicadores de Contaminación , Staphylococcus aureus/patogenicidad , Ascaris lumbricoides/parasitología , Escherichia coli/patogenicidad

RESUMEN

Abstract This study investigated the synergy testing of penicillin, cephalosporin, amphenicols, and aminoglycoside in the camel milk (n=768 samples), subsequently used for isolation of MDR S. aureus targeting mecA gene. Antibiotic susceptibility of S. aureus showed >90% isolates were sensitive to ciprofloxacin and trimethoprim and resistant against oxacillin, ampicillin, and cefoxitin. Further, 50-85% of the S. aureus were sensitive to gentamicin, oxytetracycline, and chloramphenicol and resistant against cefotaxime, vancomycin, and cefixime. Minimum inhibitory concentration (MIC) of cefotaxime, (C) and ampicillin (A) in combination with gentamicin (G) was reduced by 99.34% and 70.46%, respectively, while with chloramphenicol (Ch), reduction was 57.49% and 60%, respectively. In addition, the Fractional Inhibitory Concentration Index (FICI) of G+A, Ch+C and Ch+G combinations showed synergy against 80%, 60%, and 30% of MDR S. aureus, respectively. Similarly, C+A and Ch+G displayed indifferent interaction against 70 % and 30% of isolates, respectively, while the later showed additive interaction against 10% of MDR S. aureus. Altogether, our results described effective combination of gentamicin and chloramphenicol with ampicillin and cefotaxime to combat MDR S. aureus

Asunto(s)

Penicilinas/agonistas , Staphylococcus aureus/patogenicidad , Cloranfenicol/agonistas , Sinergismo Farmacológico , Aminoglicósidos/agonistas , Camelus/clasificación , Pruebas de Sensibilidad Microbiana/instrumentación , Genes MDR , Leche/clasificación

RESUMEN

High doses of antibiotics used in hospitals can affect the microbial composition of sewers, selecting resistant bacteria. In this sense, we evaluated the antibiotic resistance profile and the multiresistant phenotype of bacteria isolated in sewage from a tertiary hospital in the interior São Paulo state, Brazil. For bacteria isolation, 10 µL of sewage samples were sown in selective culture media and the isolates were identified using VITEK-2 automatized system. The antibiotic sensitivity test was performed by disk diffusion. High percentages of resistance were found for amoxicillin, ampicillin, ceftazidime, clindamycin, vancomycin and the multidrug-resistant phenotype (MDR) was attributed to 60.7% of the isolates. Our results show bacteria classified as critical/high priority by WHO List of Priority Pathogens (Enterococcus and Staphylococcus aureus resistant to vancomycin and Enterobacteriaceae resistant to carbapenems) in hospital sewage. Therefore, the implementation of disinfection technologies for hospital sewage would reduce the bacterial load in the sewage that will reach urban wastewater treatment plants, minimizing superficial water contamination and bacterial resistance spread in the environment.(AU)

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Altas doses de antibióticos utilizados em hospitais podem afetar a composição microbiana dos esgotos, selecionando bactérias resistentes. Nesse sentido, avaliamos o perfil de resistência a antibióticos e o fenótipo multirresistente de bactérias isoladas em esgoto de um hospital terciário no interior do estado de São Paulo, Brasil. Para o isolamento de bactérias, foram semeados 10 µL das amostras de esgoto em meios de cultura seletivos e os isolados foram identificados usando o sistema automatizado VITEK-2. O teste de sensibilidade aos antibióticos foi realizado por disco-difusão em ágar. Elevadas porcentagens de resistência foram encontradas para amoxicilina, ampicilina, ceftazidima, clindamicina, vancomicina e o fenótipo multirresistente (MDR) foi atribuído a 60,7% dos isolados. Nossos resultados mostram bactérias classificadas como prioridade crítica/alta pela Lista de Patógenos Prioritários da OMS (Enterococcus e Staphylococcus aureus resistentes à vancomicina e Enterobacteriaceae resistentes aos carbapenêmicos) no esgoto hospitalar. Sendo assim, implementação de tecnologias de desinfecção do esgoto hospitalar reduziriam a carga bacteriana no esgoto que chegará às estações de tratamento de esgoto urbanas, minimizando a contaminação dos ecossistemas hídricos receptores e a disseminação da resistência bacteriana no ambiente.(AU)

Asunto(s)

Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad , Aguas del Alcantarillado/análisis , Farmacorresistencia Microbiana , Microbiología del Agua/normas , Enterococcus/patogenicidad , Staphylococcus aureus/patogenicidad

RESUMEN

High doses of antibiotics used in hospitals can affect the microbial composition of sewers, selecting resistant bacteria. In this sense, we evaluated the antibiotic resistance profile and the multiresistant phenotype of bacteria isolated in sewage from a tertiary hospital in the interior São Paulo state, Brazil. For bacteria isolation, 10 µL of sewage samples were sown in selective culture media and the isolates were identified using VITEK-2 automatized system. The antibiotic sensitivity test was performed by disk diffusion. High percentages of resistance were found for amoxicillin, ampicillin, ceftazidime, clindamycin, vancomycin and the multidrug-resistant phenotype (MDR) was attributed to 60.7% of the isolates. Our results show bacteria classified as critical/high priority by WHO List of Priority Pathogens (Enterococcus and Staphylococcus aureus resistant to vancomycin and Enterobacteriaceae resistant to carbapenems) in hospital sewage. Therefore, the implementation of disinfection technologies for hospital sewage would reduce the bacterial load in the sewage that will reach urban wastewater treatment plants, minimizing superficial water contamination and bacterial resistance spread in the environment.

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Altas doses de antibióticos utilizados em hospitais podem afetar a composição microbiana dos esgotos, selecionando bactérias resistentes. Nesse sentido, avaliamos o perfil de resistência a antibióticos e o fenótipo multirresistente de bactérias isoladas em esgoto de um hospital terciário no interior do estado de São Paulo, Brasil. Para o isolamento de bactérias, foram semeados 10 µL das amostras de esgoto em meios de cultura seletivos e os isolados foram identificados usando o sistema automatizado VITEK-2. O teste de sensibilidade aos antibióticos foi realizado por disco-difusão em ágar. Elevadas porcentagens de resistência foram encontradas para amoxicilina, ampicilina, ceftazidima, clindamicina, vancomicina e o fenótipo multirresistente (MDR) foi atribuído a 60,7% dos isolados. Nossos resultados mostram bactérias classificadas como prioridade crítica/alta pela Lista de Patógenos Prioritários da OMS (Enterococcus e Staphylococcus aureus resistentes à vancomicina e Enterobacteriaceae resistentes aos carbapenêmicos) no esgoto hospitalar. Sendo assim, implementação de tecnologias de desinfecção do esgoto hospitalar reduziriam a carga bacteriana no esgoto que chegará às estações de tratamento de esgoto urbanas, minimizando a contaminação dos ecossistemas hídricos receptores e a disseminação da resistência bacteriana no ambiente.

Asunto(s)

Enterococcus/patogenicidad , Aguas del Alcantarillado/análisis , Microbiología del Agua/normas , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad , Farmacorresistencia Microbiana , Staphylococcus aureus/patogenicidad

RESUMEN

Introduction. Biofilm formation is a major virulence factor associated with Staphylococcus aureus infections. However, the influence of plasma proteins on biofilm formation of clinical isolates in vitro remains unclear.Hypotheses. We hypothesized that coating surfaces with plasma proteins might induce biofilm formation by S. aureus of different clonal lineages.Aim. To evaluate biofilm production by clinical S. aureus isolates of different clonal lineages isolated in Rio de Janeiro hospitals and investigated the presence of biofilm-associated genes.Methodology. This study assessed biofilm production of 60 S. aureus isolates in polystyrene microtitre plates with and without fibrinogen or fibronectin. The biochemical composition of the biofilm matrices was determined and the biofilm formation on fibrinogen-coated surfaces was also evaluated by confocal laser scanning microscopy. The presence of biofilm-related genes was detected by PCR, and the typing and functionality of agr operon was also evaluated.Results. Most of the isolates (45 %) were weak biofilm producers or non-producers. However, most of them presented a significant increase in biofilm production on plates covered with plasma proteins. There was no significant difference in biofilm formation between methicillin-resistant and -susceptible S. aureus isolates, or between different clonal lineages, except for ST30-IV (weak producers) and ST239-III (strong producers). The fnbB gene was associated with higher biofilm production.Conclusion. An increase in biofilm production in the presence of plasma proteins highlights the importance of investigating biofilm formation by S. aureus clinical isolates under different conditions since this virulence factor contributes to persistent infections and increased resistance to antimicrobials.

Asunto(s)

Biopelículas/crecimiento & desarrollo , Fibrinógeno , Fibronectinas , Staphylococcus aureus Resistente a Meticilina/patogenicidad , Staphylococcus aureus/patogenicidad , Adhesinas Bacterianas/genética , Adhesión Bacteriana/genética , Proteínas Bacterianas/genética , Genes Bacterianos , Genotipo , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Staphylococcus aureus Resistente a Meticilina/fisiología , Operón , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/genética , Staphylococcus aureus/aislamiento & purificación , Staphylococcus aureus/fisiología , Transactivadores/genética

RESUMEN

Introducción: Al colocar el catéter venoso central en enfermos de insuficiencia renal crónica durante la hemodiálisis, la prevención es imprescindible para evitar la endocarditis infecciosa. El estafilococo dorado es un germen agresivo, que en enfermos inmunodeprimidos con fenómenos cardioembólicos pulmonares y sistémicos, ocasiona daños a funciones de órganos y sistemas. El fenómeno de fallo multiórganos es una complicación temida. Objetivo: Presentar un caso de endocarditis infecciosa agresiva, en un paciente en hemodiálisis. Caso clínico: Paciente femenina, de 31 años de edad, con diagnóstico de endocarditis infecciosa, con tratamiento oportuno, adecuado y multidisciplinario. Después de una mejoría, pasó a un deterioro marcado, falleció por fallo multiórganos, debido a septicemia y cardioembolismos múltiples. Comentarios: La resistencia de los gérmenes agresivos, se hace más frecuente. La vida de la enferma, inmunodeprimida y manipulada, se sitúa en riesgo significativo con fallo multiórganos(AU)

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Introduction: When placing the central venous catheter in patients with chronic renal failure during hemodialysis, prevention is essential to avoid infective endocarditis. Staphylococcus aureus is an aggressive germ, which in immunocompromised patients with pulmonary and systemic cardioembolic phenomena, causes damage to functions of organs and systems. The phenomenon of multi-organ failure is a feared complication. Objective: To present a case of aggressive infective endocarditis in a hemodialysis patient. Clinical case: Female patient, 31 years old, with a diagnosis of infective endocarditis, with timely, adequate and multidisciplinary treatment. After an improvement, he went on to a marked deterioration, died due to multi-organ failure, due to septicemia and multiple cardioembolisms. Comment: The resistance of aggressive germs becomes more frequent. The life of the patient, immunocompromised and manipulated, is at significant risk with multi-organ failure (AU)

Asunto(s)

Humanos , Femenino , Adulto , Diálisis Renal/métodos , Endocarditis , Catéteres Venosos Centrales/efectos adversos , Fallo Renal Crónico/mortalidad , Staphylococcus aureus/patogenicidad

RESUMEN

AIM: This study aimed to assess an ex situ model of biofilm-associated wounds on porcine skin for the study of Staphylococcus aureus and Pseudomonas aeruginosa biofilms in a host-like environment, after 48 to 120 h of incubation. MATERIAL AND RESULTS: Ex situ and in vitro biofilms were comparatively analysed. Overall, CFU-counts and matrix quantification yielded significantly (P < 0·05) higher results for ex situ than in vitro biofilms. Confocal microscopy revealed greater (P < 0·05) biomass and thickness at 48-72 h and greater (P < 0·05) robustness at 72 h of growth. S. aureus ex situ biofilms produced less (P < 0·05) siderophore and proteases than in vitro biofilms, while P. aeruginosa ex situ biofilms produced more (P < 0·05) siderophores and less proteases than in vitro biofilms. CONCLUSIONS: Biofilms grown ex situ present a greater amount of bacterial cells and polymeric matrix than their in vitro counterparts, reaching maturity at 72 h of growth. Moreover the production of virulence factors differs between ex situ and in vitro biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings emphasize the importance of using ex situ biofilm models, once they mimic in vivo conditions. The use of these models brings perspectives for the pursuit of therapeutic alternatives, as tests may be performed in a host-like environment.

Asunto(s)

Biopelículas , Pseudomonas aeruginosa , Piel/microbiología , Staphylococcus aureus , Animales , Técnicas In Vitro , Infecciones por Pseudomonas , Pseudomonas aeruginosa/crecimiento & desarrollo , Pseudomonas aeruginosa/patogenicidad , Infecciones Estafilocócicas , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus aureus/patogenicidad , Porcinos , Heridas y Lesiones/microbiología

RESUMEN

Salicylic acid (SAL) has recently been shown to induce biofilm formation in Staphylococcus aureus and to affect the expression of virulence factors. This study was aimed to investigate the effect of SAL on the regulatory agr system and its impact on S. aureus biofilm formation. The agr quorum-sensing system, which is a central regulator in S. aureus pathogenicity, plays a pivotal role in the dispersal of S. aureus mature biofilms and contributes to the creation of new colonization sites. Here, we demonstrate that SAL impairs biofilm dispersal by interfering with agr expression. As revealed by our work, protease and surfactant molecule production is diminished, and bacterial cell autolysis is also negatively affected by SAL. Furthermore, as a consequence of SAL treatment, the S. aureus biofilm matrix revealed the lack of extracellular DNA. In silico docking and simulation of molecular dynamics provided evidence for a potential interaction of AgrA and SAL, resulting in reduced activity of the agr system. In conclusion, SAL stabilized the mature S. aureus biofilms, which may prevent bacterial cell dissemination. However, it may foster the establishment of infections locally and consequently increase bacterial persistence leading to therapeutic failure.

Asunto(s)

Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Ácido Salicílico/farmacología , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus/patogenicidad , Antibacterianos/uso terapéutico , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Farmacorresistencia Bacteriana , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Humanos , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Percepción de Quorum/efectos de los fármacos , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/efectos de los fármacos , Transactivadores/genética , Transactivadores/metabolismo , Factores de Virulencia/genética , Factores de Virulencia/metabolismo

RESUMEN

AIMS: This study aimed to investigate the potential of limonene as an efflux pump (EP) inhibitor in Staphylococcus aureus strains, RN-4220 and IS-58, which carry EPs for erythromycin (MrsA) and tetracycline (TetK), respectively. BACKGROUND: The evolution of bacterial resistance mechanisms over time has impaired the action of most classes of antibiotics. Staphylococcus aureus is a notable bacterium, with high pathogenic potential and demonstrated resistance to conventional antibiotics. Considering the importance of discovering novel compounds to combat antibiotic resistance, our group previously demonstrated the antibacterial properties of limonene, a compound present in the essential oils of several plant species. OBJECTIVE: This study aimed to investigate the potential of limonene as an efflux pump (EP) inhibitor in Staphylococcus aureus strains RN-4220 and IS-58, which carry EPs for erythromycin (MrsA) and tetracycline (TetK), respectively. METHODS: The minimum inhibitory concentrations (MIC) of limonene and other efflux pump inhibitors were determined through the broth microdilution method. A reduction in the MIC of ethidium bromide was used as a parameter of EP inhibition. RESULT: While limonene was not shown to exhibit direct antibacterial effects against EP-carrying strains, in association with ethidium bromide and antibiotics, this compound demonstrated enhanced antibacterial activity, indicating the inhibition of the MrsA and TetK pumps. CONCLUSION: In conclusion, this pioneering study demonstrated the effectiveness of limonene as an EP inhibitor in S. aureus strains, RN-4220 and IS-58. Nevertheless, further studies are required to characterize the molecular mechanisms associated with limonene-mediated EP inhibition.

Asunto(s)

Inhibidores Enzimáticos/farmacología , Etidio/farmacología , Limoneno/farmacología , Staphylococcus aureus , Antibacterianos/farmacología , Proteínas Bacterianas/antagonistas & inhibidores , Productos Biológicos/farmacología , Interacciones Farmacológicas , Farmacorresistencia Microbiana/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Simulación del Acoplamiento Molecular , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/patogenicidad , Staphylococcus aureus/fisiología

RESUMEN

OBJECTIVE: The present report describes the enzymatic acylation of umbelliferone with different vinyl esters as acyl donors biocatalyzed by the commercial lipase Novozym® 435, and the investigation for their antibacterial activity against ATCC and clinical strains isolated from hospital infection sites. RESULTS: The umbelliferone esters (1-5) were synthesized through the acylation reaction of 7-hydroxy-2H-chromen-2-one with different long chain vinyl esters catalyzed by the lipase Novozym 435. The reaction conditions were: 10% Novozym 435; tetrahydrofuran:acetone (3:1) for the reactions with acetate, propionate and butyrate vinyl esters 50-90% conversion, and (9:1) for decanoate and laurate vinyl esters 10-15% conversion; acyl donor/umbelliferone molar ratio of 10:1 and 60 °C. All the umbelliferone esters were characterized NMR and (HRMS). The antibacterial activity of the products were tested using the broth microdilution method in order to determine the minimum inhibitory concentration (MIC). The results displayed by 7-laurate and 7-decanoate-umbelliferone esters showed the highest antibacterial potential, with 1 mM inhibitory activity for ATCC 33591, a methicillin and oxacillin resistant Staphylococcus aureus strain. They were also able to inhibit gram-negative bacterial strains, such as Pseudomonas aeruginosa (MIC 0.5 mM) and Klebsiella pneumoniae (MIC 1 mM). In addition, 7-laurate- and 7-decanoate-umbelliferone esters were able to inhibit all clinical strains (MIC 1 mM; except 7-laurate-umbelliferone in which MIC 0.5 mM against 55a). CONCLUSIONS: This is the first study performing the biocatalysis of umbelliferone followed by the purification of the products and the antibacterial evaluation.

Asunto(s)

Infecciones Bacterianas/tratamiento farmacológico , Ésteres/farmacología , Lipasa/química , Umbeliferonas/farmacología , Antibacterianos/síntesis química , Antibacterianos/farmacología , Infecciones Bacterianas/microbiología , Biocatálisis , Enzimas Inmovilizadas/química , Enzimas Inmovilizadas/genética , Ésteres/síntesis química , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Humanos , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/patogenicidad , Lipasa/genética , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/patogenicidad , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/patogenicidad , Transducción de Señal/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/patogenicidad , Umbeliferonas/síntesis química

RESUMEN

AIMS: To evaluate the anti-staphylococcal effects of lectins isolated from bark (MuBL), heartwood (MuHL) and leaves (MuLL) of Myracrodruon urundeuva. METHODS AND RESULTS: The lectins were evaluated for: effects on growth, aggregation, haemolytic activity and biofilm-forming ability of Staphylococcus aureus clinical isolates nonresistant (8325-4) and multidrug resistant (LAC USA300); interference with the expression of virulence genes (hla, rnaIII and spa) of the Agr system of S. aureus; and synergistic effect with the antibiotics cefoxitin and cefotaxime. MuBL, MuHL and MuLL reduced growth (minimal inhibitory concentration (MIC): 12·5-50 µg ml-1 ) and viability (minimal bactericidal concentration (MBC): 100 µg ml-1 ) of 8325-4 and LAC USA300 cells. MuLL (at ½MIC and MIC) reduced LAC USA300 agglutination. The lectins did not interfere with haemolytic activity and expression of hla, rnaIII and spa genes. Only MuHL was able to reduce the biofilm production by 8325-4 (50-400 µg ml-1 ) and LAC USA300 (400 µg ml-1 ). CONCLUSION: The M. urundeuva lectins showed antibacterial activity against nonresistant and resistant clinical isolates of S. aureus and synergistic effects with antibiotics in reducing growth and biofilm formation. SIGNIFICANCE AND IMPACT OF THE STUDY: This work reports bioactive molecules capable of acting as anti-staphylococcal agents, since there are increasing reports of multiresistant isolates of this bacterium.

Asunto(s)

Anacardiaceae/química , Antibacterianos/farmacología , Lectinas de Plantas/farmacología , Staphylococcus aureus/efectos de los fármacos , Pruebas de Aglutinación , Antibacterianos/aislamiento & purificación , Biopelículas/efectos de los fármacos , Farmacorresistencia Bacteriana Múltiple , Sinergismo Farmacológico , Hemólisis/efectos de los fármacos , Humanos , Lectinas de Plantas/aislamiento & purificación , Staphylococcus aureus/aislamiento & purificación , Staphylococcus aureus/patogenicidad , Virulencia/efectos de los fármacos

RESUMEN

Light sensing has been extensively characterized in the human pathogen Acinetobacter baumannii at environmental temperatures. However, the influence of light on the physiology and pathogenicity of human bacterial pathogens at temperatures found in warm-blooded hosts is still poorly understand. In this work, we show that Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa (ESKAPE) priority pathogens, which have been recognized by the WHO and the CDC as critical, can also sense and respond to light at temperatures found in human hosts. Most interestingly, in these pathogens, light modulates important pathogenicity determinants as well as virulence in an epithelial infection model, which could have implications in human infections. In fact, we found that alpha-toxin-dependent hemolysis, motility, and growth under iron-deprived conditions are modulated by light in S. aureus Light also regulates persistence, metabolism, and the ability to kill competitors in some of these microorganisms. Finally, light exerts a profound effect on the virulence of these pathogens in an epithelial infection model, although the response is not the same in the different species; virulence was enhanced by light in A. baumannii and S. aureus, while in A. nosocomialis and P. aeruginosa it was reduced. Neither the BlsA photoreceptor nor the type VI secretion system (T6SS) is involved in virulence modulation by light in A. baumannii Overall, this fundamental knowledge highlights the potential use of light to control pathogen virulence, either directly or by manipulating the light regulatory switch toward the lowest virulence/persistence configuration.IMPORTANCE Pathogenic bacteria are microorganisms capable of producing disease. Dangerous bacterial pathogens, such as Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii, are responsible for serious intrahospital and community infections in humans. Therapeutics is often complicated due to resistance to multiple antibiotics, rendering them ineffective. In this work, we show that these pathogens sense natural light and respond to it by modulating aspects related to their ability to cause disease; in the presence of light, some of them become more aggressive, while others show an opposite response. Overall, we provide new understanding on the behavior of these pathogens, which could contribute to the control of infections caused by them. Since the response is distributed in diverse pathogens, this notion could prove a general concept.

Asunto(s)

Acinetobacter baumannii/patogenicidad , Pseudomonas aeruginosa/patogenicidad , Staphylococcus aureus/patogenicidad , Factores de Virulencia/efectos de la radiación , Acinetobacter baumannii/efectos de la radiación , Infecciones Bacterianas/microbiología , Epitelio/microbiología , Células HaCaT , Hemólisis/efectos de la radiación , Humanos , Luz , Modelos Biológicos , Pseudomonas aeruginosa/efectos de la radiación , Staphylococcus aureus/efectos de la radiación , Virulencia/efectos de la radiación

RESUMEN

Infectious diseases and the rapid development of pathogens resistant to conventional drugs are a serious global public health problem, which motivates the search for new pharmacological agents. In this context, cationic peptides without disulfide bridges from different species of scorpion venom have been the target of scientific studies due to their multifunctional activities. Stigmurin is a linear peptide composed of 17 amino acid residues (Phe-Phe-Ser-Leu-Ile-Pro-Ser-Leu-Val-Gly-Gly-Leu-Ile-Ser-Ala-Phe-Lys-NH2), which is present in the venom gland of the scorpion Tityus stigmurus. Here we present investigations of the in vitro antioxidant action of Stigmurin together with the in vivo antibacterial and healing activity of this peptide in a wound infection model induced by Staphylococcus aureus. In addition, we have reports for the first time of the three-dimensional structure determined by NMR spectroscopy of a peptide without disulfide bridges present in scorpion venom from the Tityus genus. Stigmurin showed hydroxyl radical scavenging above 70 % at 10 µM and antibiotic action in the skin wound, reducing the number of viable microorganisms by 67.2 % on the 7 day after infection. Stigmurin (1 µg / µL) increased the retraction rate of the lesion, with wound area reduction of 43 % on the second day after skin injury, which indicates its ability to induce tissue repair. Stigmurin in trifluoroethanol:water exhibited a random conformation at the N-terminus region (Phe1 to Pro6), with a helical structure from Ser7 to Phe16. This structural information, allied with the multifunctional activity of Stigmurin, makes it an attractive candidate for the design of novel therapeutic agents.

Asunto(s)

Péptidos Catiónicos Antimicrobianos/farmacología , Venenos de Escorpión/genética , Staphylococcus aureus/efectos de los fármacos , Infección de Heridas/tratamiento farmacológico , Animales , Antibacterianos/química , Antibacterianos/farmacología , Antioxidantes/química , Antioxidantes/farmacología , Humanos , Espectroscopía de Resonancia Magnética , Conformación Proteica , Venenos de Escorpión/química , Escorpiones/química , Staphylococcus aureus/patogenicidad , Infección de Heridas/microbiología

RESUMEN

Staphylococcus aureus is one of the main agents isolated from bovine mastitis cases, characterized by lower cure rates compared to other pathogens causing this disease. This phenomenon is mainly explained by the multiresistance acquisition to antimicrobials and the ability of S. aureus to form biofilms on biotic and abiotic surfaces. In this work 15 samples of S. aureus isolated from the automated milking facility were analyzed regarding the resistance profile to antimicrobials, virulence factors (capsule production, hemolysin, and protease) and adhesion capacity under different temperatures (42±1°C, 36±1°C, 25±1°C, 9±1°C, and 3±1°C). All isolates showed methicillin-resistant (MRSA) characteristics and multidrug resistance profile to the antimicrobials tested (penicillin G, chloramphenicol, oxacillin, cephalexin, tetracycline, amoxicillin + clavulanic acid, sulfa + trimetropim, gentamicin, doxycycline, ceftiofur, neomycin, and vancomycin) with an IRMA index between 0.5 and 1.0. Five isolates were resistant to vancomycin (VRSA), two were resistant to all active principles, and the others to at least six of these drugs. Adhesion capacity and biofilm formation were found in 3 of the 5 evaluated temperatures, including the cooling conditions. Regarding the virulence factors, 86.7% of the isolates formed capsules, 60% revealed the presence of protease, 26.7% expressed the α-hemolysin factor, and 13.3% of them presented β-hemolysin. The fact that all isolates presented MRSA characteristics represents a potential risk to those exposed to this agent, and the formation of biofilm in liners even after the use of detergents and sanitization highlights the urgency of searching for alternatives for dispersion of the biofilm by S. aureusin the automated milking facility.

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O Staphylococcus aureus é um dos principais agentes isolados de casos de mastite bovina, caracterizado por menores taxas de cura em comparação com outros patógenos desta enfermidade. Esse fenômeno é explicado principal-mente pela aquisição de resistência à antimicrobianos e a capacidade do S. aureus formar biofilmes em superficies bióti-cas e abióticas. Neste trabalho foram utilizadas 15 amostras de S. aureus isolados de ordenhadeira, analisados quanto ao perfil de resistência à antimicrobianos, fatores de virulência (produção de cápsula, hemolisina e protease) e capacidade de adesão sob diferentes temperaturas (42±1°C, 36±1°C, 25±1ºC, 9±1ºC e 3±1ºC). Todos os isolados apresentaram perfil de multirresistência aos antimicrobianos testados (penicilina G, cloranfenicol, oxacilina, cefalexina, tetraciclina, amoxicilina + ácido clavulônico, sulfa + trimetropim, gentamicina, doxiciclina, ceftiofur, neomicina e vancomicina) com índice IRMA entre 0,5 a 1,0. Duas cepas foram resistentes a todos os princípios ativos e as demais a pelo menos seis destes fármacos. Os isolados avaliados apresentaram característica de meticilina-resistentes (MRSA) e destes, 33,34% (5/15) foram resistentes à vancomicina (VRSA). Houve capacidade de adesão e formação de biofilmes em 3 das 5 tem-peraturas avaliadas, incluindo as temperaturas de refrigeração. Em relação aos fatores de virulência, 86,7% dos isolados formaram cápsula, 60% presença de protease, 26,7% expressaram o fator α-hemolisina e 13,3% β-hemolisina. O fato de todos isolados apresentarem característica MRSA representa um risco potencial aos expostos a esse agente. Já a for-mação de biofilmes em teteiras, mesmo após detergência e sanitização, destacam a urgência de alternativas de dispersão de biofilmes no ambiente de ordenha.

Asunto(s)

Antiinfecciosos/análisis , Biopelículas , Staphylococcus aureus/inmunología , Staphylococcus aureus/patogenicidad , Mastitis Bovina

RESUMEN

O objetivo do trabalho foi avaliar a atividade antimicrobiana in vitro de diferentes óleos essenciais e suas combinações contra linhagens patogênicas de Salmonella typhimurium ATCC 13311 e Staphylococcus aureus ATCC 13565. Foram testados ao todo 41 óleos essenciais e as combinações em pares dos óleos de alecrim-pimenta (Lippia sidoides Cham.), capim-limão (Cymbopogon citratus Stapf.), tomilho (Thymus vulgaris L.), orégano (Origanum vulgare L.) e canela (Cinnamomum cassia (L.) D.Don. para observar efeitos adicionais ou não na inibição dos microrganismos. Os resultados mostraram que as melhores atividades antimicrobianas para os dois microrganismos foram obtidas utilizando os óleos de Thymus vulgaris L. (tomilho), Origanum vulgare L. (orégano) e Lippia sidoides Cham. (alecrim-pimenta). As combinações de cada par dos óleos de alecrim-pimenta, capim-limão, tomilho, orégano e canela não apresentaram uma melhora adicional no efeito para os patógenos estudados quando comparados com os seus óleos testados isoladamente. As composições químicas dos óleos essenciais mais ativos foram obtidas por meio de análises por cromatografia gasosa acoplada à espectrometria de massas (CG-EM), sendo que os compostos majoritários do óleo de orégano foram carvacrol (69,1%) e p-cimeno (18,8%), do óleo essencial de tomilho foram timol (45,5%) e p-cimeno (35,6%) e do alecrim-pimenta foram o timol (77,2%) e p-cimeno (14,2%). Análises quantitativas de p-cimeno, carvacrol e timol nos três óleos foram realizadas e comparadas com as composições químicas mostrando resultados proporcionalmente coerentes, exceto para p-cimeno, que foi quantificado com menores porcentagens por esse método. A aplicabilidade dos óleos selecionados, ou das substâncias presentes isoladas seria diretamente nos alimentos, com a finalidade de inibir ou controlar o crescimento desses patógenos, contribuindo assim para a segurança dos alimentos. (AU)

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The aim of this study was to evaluate the in vitro antimicrobial activity of several essential oils and their combination against pathogenic strains of Salmonella typhimurium ATCC 13311 and Staphylococcus aureus ATCC 13565. A total of 41 essential oils and the combinations in pairs of the oils of rosemary pepper (Lippia sidoides Cham.), lemongrass (Cymbopogon citratus Stapf.), thyme (Thymus vulgaris L.), origanum (Origanum vulgare L.) and cinnamon (Cinnamomum cassia (L.) D.Don) were tested to observe additional effects or not in the inhibition of microorganisms. The results showed that the best antimicrobial activities for the two microorganisms were obtained using the oils of Thymus vulgaris L. (thyme), Origanum vulgare L. (origanum) and Lippia sidoides Cham. (rosemary pepper). The combinations of each pair of rosemary pepper, lemongrass, thyme, origanum and cinnamon oils did not show any further improvement in the effect for the studied pathogens when compared to their oils tested alone. The compositions of the most active essential oils were obtained by gas chromatography coupled to mass spectrometry (GC-MS). The major compounds in origanum oil were carvacrol (69.1%) and p-cymene (18.8%), thyme essential oil were thymol (45.5%) and p-cymene (35.6%) and rosemary pepper were thymol (77.2%) and p-cymene (14.2%). Quantitative analyses of p-cymene, carvacrol and thymol confirmed the presence in thyme, oregano and pepper rosemary oils and were compared with the chemical composition showing similar results, except for p-cymene, which was quantified with lower percentages in these oils. One of the possible applications of the results of this work is the use of essential oils, or the isolated substances, directly in foods, with the purpose of inhibiting or controlling the growth of these pathogens, thus contributing to the food safety.Keywords: Bioassay. GC-MS. Inhibition. Carvacrol. Thymol. p-cymene(AU)

Asunto(s)

Salmonella typhimurium/patogenicidad , Staphylococcus aureus/patogenicidad , Aceites Volátiles , Antiinfecciosos/química

RESUMEN

Staphylococcus aureus is one of the main agents isolated from bovine mastitis cases, characterized by lower cure rates compared to other pathogens causing this disease. This phenomenon is mainly explained by the multiresistance acquisition to antimicrobials and the ability of S. aureus to form biofilms on biotic and abiotic surfaces. In this work 15 samples of S. aureus isolated from the automated milking facility were analyzed regarding the resistance profile to antimicrobials, virulence factors (capsule production, hemolysin, and protease) and adhesion capacity under different temperatures (42±1°C, 36±1°C, 25±1°C, 9±1°C, and 3±1°C). All isolates showed methicillin-resistant (MRSA) characteristics and multidrug resistance profile to the antimicrobials tested (penicillin G, chloramphenicol, oxacillin, cephalexin, tetracycline, amoxicillin + clavulanic acid, sulfa + trimetropim, gentamicin, doxycycline, ceftiofur, neomycin, and vancomycin) with an IRMA index between 0.5 and 1.0. Five isolates were resistant to vancomycin (VRSA), two were resistant to all active principles, and the others to at least six of these drugs. Adhesion capacity and biofilm formation were found in 3 of the 5 evaluated temperatures, including the cooling conditions. Regarding the virulence factors, 86.7% of the isolates formed capsules, 60% revealed the presence of protease, 26.7% expressed the α-hemolysin factor, and 13.3% of them presented β-hemolysin. The fact that all isolates presented MRSA characteristics represents a potential risk to those exposed to this agent, and the formation of biofilm in liners even after the use of detergents and sanitization highlights the urgency of searching for alternatives for dispersion of the biofilm by S. aureusin the automated milking facility.(AU)

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O Staphylococcus aureus é um dos principais agentes isolados de casos de mastite bovina, caracterizado por menores taxas de cura em comparação com outros patógenos desta enfermidade. Esse fenômeno é explicado principal-mente pela aquisição de resistência à antimicrobianos e a capacidade do S. aureus formar biofilmes em superficies bióti-cas e abióticas. Neste trabalho foram utilizadas 15 amostras de S. aureus isolados de ordenhadeira, analisados quanto ao perfil de resistência à antimicrobianos, fatores de virulência (produção de cápsula, hemolisina e protease) e capacidade de adesão sob diferentes temperaturas (42±1°C, 36±1°C, 25±1ºC, 9±1ºC e 3±1ºC). Todos os isolados apresentaram perfil de multirresistência aos antimicrobianos testados (penicilina G, cloranfenicol, oxacilina, cefalexina, tetraciclina, amoxicilina + ácido clavulônico, sulfa + trimetropim, gentamicina, doxiciclina, ceftiofur, neomicina e vancomicina) com índice IRMA entre 0,5 a 1,0. Duas cepas foram resistentes a todos os princípios ativos e as demais a pelo menos seis destes fármacos. Os isolados avaliados apresentaram característica de meticilina-resistentes (MRSA) e destes, 33,34% (5/15) foram resistentes à vancomicina (VRSA). Houve capacidade de adesão e formação de biofilmes em 3 das 5 tem-peraturas avaliadas, incluindo as temperaturas de refrigeração. Em relação aos fatores de virulência, 86,7% dos isolados formaram cápsula, 60% presença de protease, 26,7% expressaram o fator α-hemolisina e 13,3% β-hemolisina. O fato de todos isolados apresentarem característica MRSA representa um risco potencial aos expostos a esse agente. Já a for-mação de biofilmes em teteiras, mesmo após detergência e sanitização, destacam a urgência de alternativas de dispersão de biofilmes no ambiente de ordenha.(AU)

Asunto(s)

Staphylococcus aureus/inmunología , Staphylococcus aureus/patogenicidad , Antiinfecciosos/análisis , Biopelículas , Mastitis Bovina