Characterisation and anti-biofilm activity of glycerol monolaurate nanocapsules against Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a ubiquitous microorganism that commonly causes hospital-acquired infections, including pneumonia, bloodstream and urinary tract infections and it is well known for chronically colonising the respiratory tract of patients with cystic fibrosis, causing severe intermittent exacerbation of the condition. P. aeruginosa may appear in the free form cell but also grows in biofilm communities adhered to a surface. An alternative to conventional antimicrobial agents are nanoparticles that can act as carriers for antibiotics and other drugs. In this context, the study aimed to characterise and verify the anti-biofilm potential of GML Nanocapsules against P. aeruginosa. The nanocapsules showed a mean diameter of 190.7 nm, polydispersion index of 0.069, the zeta potential of -23.3 mV. The microdilution test showed a MIC of 62.5 µg/mL to GML and 15.62 µg/mL to GML Nanocapsules. The anti-biofilm experiments demonstrated the significant reduction of biomass, proteins, polysaccharide and viable P. aeruginosa in biofilm treated with GML Nanocapsules while the free GML did not cause an effect. The AFM images showed a decrease in a biofilm which received GML. The positive results suggest an alternative for the public health trouble related to infections associated with biofilm.

Antimicrobial activity and phytochemical characterization of Carya illinoensis.

Carya illinoensis is a widespread species, belonging to the Juglandaceae family, commonly known as Pecan. Popularly, the leaves have been used in the treatment of smoking as a hypoglycemic, cleansing, astringent, keratolytic, antioxidant, and antimicrobial agent. The following research aimed to identify for the first time the phytochemical compounds present in the leaves of C. illinoensis and carry out the determination of antimicrobial activity of aqueous and ethanolic extracts. The antimicrobial activity was tested against 20 microorganisms by determining the minimum inhibitory concentration (MIC). Phenolic acids (gallic acid and ellagic acid), flavonoids (rutin), and tannins (catechins and epicatechins) were identified by HPLC-DAD and may be partially responsible for the antimicrobial activity against Gram-positive, Gram-negative, and yeast. The results showed MIC values between 25 mg/mL and 0.78 mg/mL. The extracts were also able to inhibit the production of germ tubes by Candida albicans.

Nanocapsules with glycerol monolaurate: Effects on Candida albicans biofilms.

Candida albicans does not only occur in the free living planktonic form but also grows in surface-attached biofilm communities. Moreover, these biofilms appear to be the most common lifestyle and are involved in the majority of human Candida infections. Nanoparticles can be used as an alternative to conventional antimicrobial agents and can also act as carriers for antibiotics and other drugs. In view of this, the aim of the study was develop, characterize and verify the anti-biofilm potential of GML Nanocapsules against C. albicans. The GML Nanocapsules showed mean diameter of 193.2 nm, polydispersion index of 0.044, zeta potential of -23.3 mV and pH 6.32. The microdilution assay showed MIC of 15.5 µg mL(-1) to GML Nanocapsules and 31.25 µg mL(-1) to GML. The anti-biofilm assay showed the significantly reduction of biomass of C. albicans biofilm treated with GML Nanocapsules while the GML does not exhibit effect. The kinetic assay demonstrated that at 48 h, the GML Nanocapsules reduce 94% of formed biofilm. The positive results suggest the promisor alternative for this public health problem that is biofilm infections.

Anti-Candida activity assessment of Pelargonium graveolens oil free and nanoemulsion in biofilm formation in hospital medical supplies.

Infections due to microbial biofilm formation on the surface of catheters and other medical devices are constantly reported as a major cause of morbidity and mortality in patients admitted to hospitals. Furthermore, sessile cells are more resistant to phagocytosis and most antimicrobial, which complicates the treatment of such infections. Researches aimed at new antimicrobial originating mainly from plants have increased in recent years and the development of new strategies for their release is critical in combating the formation of biofilms. Geranium oil (GO) has proven antimicrobial activity. Because of this, the aim of this study was to develop nanoemulsions containing this oil (NEG) and evaluate its activity after the biofilm formation of Candida albicans, Candida tropicalis, Candida glabrata, and Candida krusei in hospital medical supplies. For quantification of the biofilm, crystal violet, total protein, and ATP-bioluminescence assays were used. The results revealed that GO and NEG showed lower MIC for C. albicans and C. tropicalis. The biofilms formed by different species of Candida on the surfaces of polyethylene and polyurethane were quantified. GO and NEG significantly inhibited the formation of biofilms in all species tested on the surfaces of polyethylene. However, NEG antibiofilm has had better activity than GO for C. albicans, C. tropicalis and C. glabrata, according to the surface potential analysis by atomic force microscopy (AFM). The analysis of the biofilm formation on the polyethylene surface by ATP-bioluminescence and CFU showed similar results. In both methods the formation of biofilm in the catheter occurred in greater quantity for C. albicans and C. tropicalis. GO did not significantly inhibit the formation of biofilms only in C. krusei, although NEG significantly increased this activity GO in all species tested when compared to the control training biofilm. The following study shows that the development of NEG may become an effective alternative to reduce the adhesion of microorganisms and prevent infections resulting from the use of some hospital medical materials.

Efficacy of calcium hypochlorite in disinfection of gutta-percha cones contaminated with Candida albicans.

This study aimed to compare the efficacy of 2.5% sodium hypochlorite (NaOCl), 2.5% calcium hypochlorite [Ca(OCl)2], and 2% chlorhexidine (CHX) in the rapid disinfection of gutta-percha cones contaminated with Candida albicans. The minimum inhibitory and minimum fungicidal concentrations of each solution for C. albicans were determined and the ability of each solution to destroy and inhibit biofilm in culture wells was tested. In addition, ninety-eight gutta-percha cones contaminated with the fungal suspension were disinfected according to the type of solution (2.5% NaOCl, 2.5% Ca(OCl)2 or 2% CHX) in its different application methods (without agitation, ultrasonic agitation or agitation with Easy Clean), and regarding the exposure time to each irrigating solution (1 or 5 min). Next, the samples were checked for turbidity and evaluation of viable colonies. The compounds that showed the best performance in biofilm destruction were NaOCl and Ca(OCl)2 at a concentration of 2xMIC (p < 0.001). Regarding inhibited biofilm, the only compound that was effective at all MIC concentrations tested was 2.5% Ca(OCl)2 (p < 0.0001). Regarding the viable colonies, all solutions were effective concerning the control group, for all application methods, in 1 and 5 min (p < 0.05). The densitometer reading showed that CHX was the only effective solution in all application methods performed (p < 0.05). The results demonstrate that all tested solutions were effective in the rapid decontamination of cones contaminated with C. albicans.

Antimicrobial activity of Amazonian oils against Paenibacillus species.

The Gram-positive, spore-forming bacterium Paenibacillus larvae is the primary bacterial pathogen of honeybee brood and the causative agent of American foulbrood disease (AFB). One of the feasible alternative treatments being used for their control of this disease is essential oils. In this study in vitro antimicrobial activity of Andiroba and Copaíba essential oils against Paenibacillus species, including P. larvae was evaluated. Minimal inhibitory concentration (MIC) in Mueller-Hinton broth by the microdilution method was assessed. Andiroba registered MIC values of 1.56-25%, while the MICs values obtained for Copaíba oil were of 1.56-12.5%. In order to determine the time-response effect of essential oils on P. larvae, this microorganism was exposed to the oils for up to 48 h. After 24 h treatment with Andiroba oil and after 48 h treatment with Copaíba oil no viable cells of P. larvae ATCC 9545 were observed. The possible toxic effect of essential oils were assessed by the spraying application method of the same concentrations of MICs. Bee mortality was evident only in treatment with Andiroba oil and the Copaíba oil shows no toxic effects after 10 days of observation. Taking together ours results showed for the first time that these oils presented a high activity against Paenibacillus species showing that Copaíba oil may be a candidate for the treatment or prevention of AFB.

Detection and characterization of Bacillus cereus isolated from the dialysis fluid.

In this study, B. cereus was detected in dialysis fluids within international parameters (ultrapure - maximum limit of 0.1 CFU/mL for heterotrophic bacteria count) by analyzing the pellet obtained through the centrifugation method. We also investigated the ability of the B. cereus isolate to form a biofilm at different temperatures, the production of virulence factors, and the susceptibility to commercial antimicrobial agents. This study demonstrated a high ability of B. cereus to persist in the hemodialysis system, which can be explained by its broad ability to produce a biofilm at 25 °C, its relevant production of virulence factors, such as ß-hemolysin, lecithinase and cereulide, and its important resistance pattern to antimicrobial drugs. In conclusion, these new findings expand the understanding that this microorganism should not be neglected and new methods for tracking it should be considered.

Bioprospection of novel synthetic monocurcuminoids: Antioxidant, antimicrobial, and in vitro cytotoxic activities.

The irrational use of medications has increased the incidence of microbial infections, which are a major threat to public health. Moreover, conventional therapeutic strategies are starting to become ineffective to treat these infections. Hence, there is a need to develop and characterize novel antimicrobial compounds. Phytochemicals are emerging as a safe and accessible alternative to conventional therapeutics for treating infectious diseases. Curcumin is extracted from the dried rhizome of the spice turmeric (Curcuma longa (Zingiberaceae)). However, the bioavailability of curcumin is low owing to its lipophilic property and thus has a low therapeutic efficacy in the host. A previous study synthesized structural variants of curcumin, which are called monocurcuminoids (CNs). CNs are synthesized based on the chemical structure of curcumin with only one methyl bridge. The biological activities of four previously synthesized CNs (CN59, CN63, CN67, and CN77), curcumin, and turmeric powder were examined in this study. Gas chromatography-tandem mass spectrometry analysis of curcumin and turmeric powder revealed similar peaks, which indicated the presence of curcumin in turmeric powder. The antioxidant activity of the test compounds was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays. The ABTS radical scavenging activities of the test compounds were similar to those of vitamin C. The minimum inhibitory concentration (MIC) values of the test compounds against seven microbial strains were in the range of 4.06-150 µg/mL. The MIC value was equal to minimum bactericidal concentration value for CN63 (150 µg/mL) and CN67 (120 µg/mL) against Staphylococcus aureus. The treatment combination of CN77 (8.75 or 4.37 µg/mL) and turmeric powder (9.37 or 4.68 µg/mL) exerted synergistic growth-inhibiting effects on Aeromonas hydrophila, Candida albicans, and Pseudomonas aeruginosa. Photodynamic therapy using 2X MIC of CN59 decreased the growth of Enterococcus faecalis by 4.18-fold compared to the control group and completely inhibited the growth of Escherichia coli. The results of the hemolytic assay revealed that the test compounds were not cytotoxic with half-maximal inhibitory concentration values ranging from 49.65-130.9 µM. The anticoagulant activity of most compounds was comparable to that of warfarin but higher than that of heparin. This indicated that these compounds target the intrinsic coagulation pathway. These results demonstrated that these CNs are a safe and promising alternative for curcumin.

Toxicological evaluation of naringin-loaded nanocapsules in vitro and in vivo.

Naringin is a flavonoid widely known for its pharmacological properties, such as: anti-inflammatory and antioxidant ones, being an ally to avoid oxidative damage. Although naringin is an active easily found in citrus fruits, it has low bioavailability, biodistribution and also undergoes biotransformation in naringenin, limiting the described effects. The use of nanocapsules as drug carriers may increase solubility, improve biodistribution, impede the biotransformation thereof, and thus could improve the performance of naringin for use in treating neurological diseases. Therefore, the objective of this work is to produce a nanocapsule containing naringin, validate an analytical method by RP-HPLC to determination of the drug in nanoparticle and evaluate the toxicity. To that end, the blank nanocapsules (NB, without the drug) or naringin-loaded nanocapsules (NN) at the concentration of 2 mg/mL were prepared by interfacial deposition of the preformed polymer and the quantification of naringin by HPLC. Toxicity of the formulations was evaluated in vitro in rat hippocampal slices and in vivo models with C. elegans and Danio rerio (zebrafish). The analytical parameters evaluated (linearity, limit of detection and quantification, specificity, precision, accuracy and robustness) indicated adequate method to assay of naringin in nanocapsules by HPLC. There was no indication of toxicity by the nanocapsules in the evaluated biological assays.

Synthesis and evaluation of the antimicrobial activity of purified and unpurified multi-walled carbon nanotubes

Multi-walled carbon nanotubes (MWCNTs) were purified and unpurified in this study to obtain hybrid materials with improved activity. The production stage comprised a chemical purification of the produced sample. Raman spectroscopy analyzed the structural composition of purified and unpurified samples. The disc diffusion assay, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and time-kill assessment analyzed antimicrobial activity. MWCNT performed well against the tested bacteria (Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterobacter aerogenes, Staphylococcus aureus, and Streptococcus sp. Staphylococcus epidermidis). The disc diffusion assay revealed inhibition zone differences caused by purified and unpurified MWCNTs. MIC and MBC values of purified and unpurified MWCNTs were similar. The purified and unpurified nanotubes of Staphylococcus epidermidis (ATCC 35985) exhibited inhibition zone diameters of approximately 8 mm and 9 mm, respectively. The microdilution method revealed a MIC of 1.23 mg/ml for the purified nanotube and 0.156 mg/ml for the unpurified nanotube against the same microorganism. The killing curve analysis demonstrated that unpurified carbon nanotubes were more effective against all tested microorganisms. MWCNTs represent a promising method for microbiology, but studies on the toxicity of these materials remain scarce.

Glycerol monolaurate nanocapsules for biomedical applications: in vitro toxicological studies.

The glycerol monolaurate (GML) is a surfactant used in the food industry and has potent antimicrobial activity against many microorganisms; however, the use of GML is not expanded due its high melting point and poor solubility in water. The aim of the study was to produce, characterize, and evaluate in vitro the cytotoxicity of GML and GML nanocapsules. The GML nanocapsules were produced and characterized by a mean diameter, zeta potential, and polydispersity index. The cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH) release, thiobarbituric acid reactive substances (TBARS), and hemolytic activity. The genotoxicity was verified by comet assay. The physicochemical parameters showed a mean diameter of 192.5 ± 2.8 nm, a polydispersity index of 0.061 ± 0.018, and a zeta potential about - 21.9 ± 1 mV. The viability test demonstrated the protector effect of GML nanocapsule compared with the GML on peripheral blood mononuclear cells (PBMC) and VERO cells (isolated from kidney epithelial cells extracted from an African green monkey). A reduction in lipid peroxidation and lactate dehydrogenase release in GML nanocapsule-exposed cells compared with GML treated cells was observed. The damage on erythrocytes was addressed in treatment with GML, while the treatment with GML nanocapsules did not cause an effect. Moreover, the comet assay showed that the GML-caused genotoxicity and GML nanocapsules do not demonstrate damage. The study showed the reduction of toxicity of GML nanocapsules by many methods used in antimicrobial therapy.

The influence of different culture media on Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus biofilm formation

Microorganisms such as Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus are frequently isolated in samples of urinary, blood, intestinal, and respiratory infections, among others. These bacteria are also associated with microbial biofilm formation. E. coli, P. aeruginosa, and S. aureus biofilm infections are particularly hard to manage and often associated with nosocomial problems. This study investigated the influence of different culture media on E. coli, P. aeruginosa, and S. aureus biofilm formation. Bacterial performance was evaluated in brain heart infusion broth, Mueller-Hinton broth, or tryptic soy broth, with or without supplementing with different glucose levels (1-5%). The study quantified biofilm biomass and the count of viable biofilm colonies. This is the first study that compares the biofilm formation of E. coli, P. aeruginosa and S. aureus in polystyrene using different culture media and with different glucose concentrations. The most robust growth of E. coli, P. aeruginosa, and S. aureus occurred in brain heart infusion broth supplemented with 5% glucose, Mueller-Hinton broth without glucose, and tryptic soy broth with 2% glucose, respectively. Our data demonstrate that behavioral and morphological characteristics of each bacterium require a specific broth to enhance the growth of these microorganisms. These findings will contribute to future tests for therapeutic alternatives with anti-biofilm potential.

Acute toxicity and antimicrobial activity of leaf tincture Baccharis trimera (Less).

BACKGROUND: The present study aimed to evaluate the possible acute oral toxicity of Baccharistrimera leaf dye as well as its antimicrobial activity. METHOD: Organization for Economic co-operation and development (OECD) 423 was used to assess acute oral toxicity and as per protocol a dose of 2000 mg/kg of tincture was administered to Wistar rats, male and female, and observed for 14 days. Biochemical and hematological analyzes were performed with sample collected of rat. The dye was evaluated for antimicrobial activity by agar diffusion and microdilution methods, which allow to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) and antibiofilm potential. RESULTS: The results showed that there was no loss of animals and no significant changes in hematological and biochemical parameters after oral administration of 2000 mg/kg of tincture and was considered safe by the OECD, classified as category 5. The dyeing also showed an important antimicrobial activity against gram positive and gram negative bacteria also significantly decreased the microbial biofilm. CONCLUSION: The tincture of B.trimera leaf when given orally once can be considered safe and has a relevant antimicrobial potential that should be elucidated in subsequent research.

Detection and characterization of Bacillus cereus isolated from the dialysis fluid

ABSTRACT In this study, B. cereus was detected in dialysis fluids within international parameters (ultrapure - maximum limit of 0.1 CFU/mL for heterotrophic bacteria count) by analyzing the pellet obtained through the centrifugation method. We also investigated the ability of the B. cereus isolate to form a biofilm at different temperatures, the production of virulence factors, and the susceptibility to commercial antimicrobial agents. This study demonstrated a high ability of B. cereus to persist in the hemodialysis system, which can be explained by its broad ability to produce a biofilm at 25 °C, its relevant production of virulence factors, such as β-hemolysin, lecithinase and cereulide, and its important resistance pattern to antimicrobial drugs. In conclusion, these new findings expand the understanding that this microorganism should not be neglected and new methods for tracking it should be considered.

Evaluation of photodynamic activity, photostability and in vitro drug release of zinc phthalocyanine-loaded nanocapsules.

Nanocapsule formulations containing zinc phthalocyanine (ZnPc) were investigated as drug delivery systems for use in photodynamic therapy (PDT). ZnPc loaded chitosan, PCL, and PCL coated with chitosan nanocapsules were prepared and characterized by means of their physicochemical properties, photodynamic activity, photostability and drug release profile. All formulations presented nanometric hydrodynamic radius, around 100 nm, low polydispersity index (0.08-0.24), slightly negative zeta potential for PCL nanoparticles and positive zeta potential for suspension containing chitosan. Encapsulation efficiencies were higher than 99%. The capacity of ZnPc loaded nanocapsules to produce cytotoxic singlet oxygen ((1)O2) by irradiation with red laser was monitored using 1.3-diphenylisobenzofuran as a probe. The singlet oxygen quantum yields (ΦΔ) for ZnPc loaded chitosan nanocapsules were high and similar to that of the standard (ZnPc in DMSO), displaying excellent ability to generate (1)O2. The photosensitizer loaded nanocapsules are photostable in the timescale usually utilized in PDT and only a small photobleaching event was observed when a light dose of 610J/cm(2) was applied. The in vitro drug release studies of ZnPc from all nanocapsules demonstrated a sustained release profile controlled by diffusion, without burst effect. The nature of the polymer and the core type of the nanocapsules regulated ZnPc release. Thus, the nanocapsules developed in this work are a promising strategy to be employed in PDT.

Análise microbiológica da ocorrência da contaminação bacteriana da mesa cirúrgica comparando o TNT gramatura 20 e gramatura 40 / Microbiological analysis of occurrence of bacterial contamination of surgical table comparing 20 thickness and 40 thicknes

A limpeza e desinfecção das superfícies operatórias fixas e partes expostas do equipo odontológico reduzem, significativamente, a contaminação. Este estudo teve como objetivo avaliar se há contaminação da mesa cirúrgica, ao se utilizar o TNT esterilizado nas gramaturas 20 g/m² e 40 g/m². Materiais e Métodos: O trabalho constituiu-se de 2 grupos, compostos por 30 amostras (campos cirúrgicos) para cada grupo. Após a desinfecção prévia das mesas cirúrgicas, foram colocados campos de TNT. No grupo 1, foi utilizado TNT gramatura 20, e no grupo 2, gramatura 40. A coleta das amostras dos dois grupos foi realizada logo após a colocação dos campos cirúrgicos (tempo 1) e 1 hora após (tempo 2). Resultados: Em relação à ocorrência de contaminação bacteriana, não houve diferença estatisticamente significativas entre tempo de coletas. Mesmo com os cuidados de desinfecção da mesa e paramentação adequada, esses campos não foram totalmente eficazes. Conclusão: A amostra do TNT gramatura 40, tanto no início da cirurgia quanto 1 hora após, obteve melhores resultados, comparando-se com o TNT gramatura 20 sem diferenças estatísticas... (AU)

The cleaning and disinfecting fixed operative surfaces and exposed parts of the dental unit significantly reduces contamination. This study aimed to assess whether there is contamination of the surgical table to use the TNT sterilized in the weights 20 g/m² and 40 g/m². Materials and Methods: The study consisted of two groups, composed of 30 samples (drapes) for each group. After prior disinfection of surgical tables were placed TNT fields. Group 1 was used TNT weight and 20 in group 2 weight 40. The sample collection from both groups was held soon after the placement of surgical fields (time 1) and 1 hour after (time 2). Results: Regarding the occurrence of bacterial contamination, there was no statistically significant difference between time of collection. Even with the disinfecting care of the table and adequate scrub, these fields have not been fully effective. Conclusion: Thus, the sample weight of the TNT 40, both at the beginning of surgery, as 1 hour, better results compared to the TNT weight 20 without statistical differences... (AU)