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1.
Pathogens ; 12(5)2023 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-37242370

RESUMO

The agar dilution method (ADM) recommended for IV fosfomycin (IV FOS) is complex and labor-intensive. Keeping in mind the reality of everyday laboratory work, we have evaluated the agreement of IV FOS susceptibility results obtained using the E-test and the Phoenix system with the results obtained using the ADM. MATERIALS AND METHODS: The tests were performed on 860 strains. To evaluate susceptibility to IV FOS, BioMerieux E-tests (bioMerieux, Warsaw, Poland), BD Phoenix panels (BD Phoenix, Sparks, MD, USA), and the ADM were used. Clinical interpretation was performed in accordance with EUCAST Guidance (v12.0, 2021). The significance of the E-test and the Phoenix was analyzed in relation to the ADM by defining categorical agreement (CA), major error (ME), and very major error (VME). Essential agreement (EA) has also been defined for the E-test. A method was considered reliable, in accordance with ISO 20776-2:2007, when CA and EA were above 89.9% and VME was <3%. RESULTS: A categorical agreement of >98.9% was demonstrated between the E-test and the ADM for overall strains and for Echerichia coli, ESBL-producing Enterobacterales, and Staphylococcus aureus, while between the Phoenix and the ADM, a CA of >98.9% was shown only for Escherichia coli, Staphylococcus aureus, and Proteus spp. A very major error rate of <3% was obtained only for Staphylococcus aureus and MBL-producing Pseudomonas evaluated by both the E-test and the Phoenix. An essential agreement of >98.9% between the E-test and the ADM has not been demonstrated for any of the tested groups of strains. The Phoenix yielded more VMEs than the E-test (50 and 46, respectively). The highest VME rate was demonstrated using the Phoenix method for Enterobacter spp. (53.83%). CONCLUSIONS: Both the E-test and the Phoenix have turned out to be reliable in assessing IV FOS susceptibility only for Staphylococcus aureus (CA > 89.9% and VME < 3%). For the remaining tested groups of strains and genera, the simultaneous high CA rate and low VME rate required by ISO were not achieved. Both methods fared particularly badly in detecting strains resistant to IV.

2.
Pathogens ; 11(12)2022 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-36558775

RESUMO

Multidrug resistance of bacteria has prompted intensive development work on new medicines, but also the search for effective options among the oldest antibiotics. Although intravenous fosfomycin (IVFOS) seems to be an interesting proposal, the recommended agar dilution method for susceptibility determination poses a major problem in routine diagnostic testing. As a consequence, there is a lack of comprehensive data on the frequency of isolation of susceptible or resistant strains. This fact triggered the disposition of EUCAST concerning the revision of IVFOS breakpoints (BPs), including withdrawal of BPs for Enterobacterales (excluding E. coli) and coagulase-negative staphylococci. Therefore, the aim of this study was to assess the activity of fosfomycin against numerous clinical strains using recommended methods. Materials and methods: A total of 997 bacterial strains were tested from the following genera: Enterobacterales, Pseudomonas spp., Staphylococcus spp., Acinetobacter spp., and Enterococcus spp., for which there are currently no BPs. The strains were isolated from various clinical materials from patients hospitalized in five hospitals. During the investigation, the recommended agar dilution method was used. Susceptibility to other antibiotics and resistance mechanisms were determined using an automatic method (Phoenix) the disk diffusion method, and E-tests. MIC values of fosfomycin were estimated for all strains and for susceptible and multidrug-resistant (MDR) strains individually. Results: Except for Acinetobacter and Enterococcus, 83% of the strains were susceptible to IVFOS, including the largest percentage of S. aureus and E. coli. Klebsiella spp. turned out to be the least susceptible strains (66%). The highest proportion of susceptibility to fosfomycin was found among strains that were sensitive to other antibiotics (80.9%), and the lowest was found among Gram-negative carbapenemase-producing bacteria (55.6%) and ESBL+ bacteria (61.6%). The MIC evaluation revealed the lowest MIC50 and MIC90 values for S. aureus (0.5 mg/L and 1 mg/L, respectively) and E. coli (4 mg/L and 32 mg/L, respectively). The highest values of MIC50 were found for Acinetobacter spp. (256 mg/L), while the highest values of MIC90 were found for Acinetobacter spp. and Klebsiella spp. (256 mg/L and 512 mg/L, respectively). Conclusions: IVFOS appears to be suitable for the treatment of many infections, including the empirical treatment of polymicrobial infections and those caused by MDR strains, since the sensitivity of the studied strains to this antibiotic in different groups ranged from 66% to as much as 99%. Sensitivity to fosfomycin was also demonstrated by 60% of carbapenem-resistant strains; therefore, IVFOS is one of the few therapeutic options that can be effective against the most resistant Gram-negative rods. In light of the general consultation posted by EUCAST, obtaining data such as IVFOS MIC value distributions may be vital for the decision of implementing fosfomycin into breakpoint tables.

3.
Pathogens ; 9(5)2020 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-32455691

RESUMO

Bacterial wound infections are a common problem associated with surgical interventions. In particular, biofilm-forming bacteria are hard to eradicate, and alternative methods of treatment based on covering wounds with vascularized flaps of tissue are being developed. The greater omentum is a complex organ covering the intestines in the abdomen, which support wound recovery following surgical procedures and exhibit natural antimicrobial activity that could improve biofilm eradication. We investigated changes in rats' metabolome following Klebsiella pneumoniae infections, as well as the greater omentum's ability for Klebsiella pneumoniae biofilm eradication. Rats received either sterile implants or implants covered with Klebsiella pneumoniae biofilm (placed in the peritoneum or greater omentum). Metabolic profiles were monitored at days 0, 2, and 5 after surgery using combined proton nuclear magnetic resonance (1H NMR) and high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF­MS) measurements of urine samples followed by chemometric analysis. Obtained results indicated that grafting of the sterile implant to the greater omentum did not cause major disturbances in rats' metabolism, whereas the sterile implant located in the peritoneum triggered metabolic perturbations related to tricarboxylic acid (TCA) cycle, as well as choline, tryptophan, and hippurate metabolism. Presence of implants colonized with Klebsiella pneumoniae biofilm resulted in similar levels of metabolic perturbations in both locations. Our findings confirmed that surgical procedures utilizing the greater omentum may have a practical use in wound healing and tissue regeneration in the future.

4.
Toxins (Basel) ; 11(7)2019 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-31336994

RESUMO

Chelidonium majus (Papaveraceae) extracts exhibit antimicrobial activity due to the complex alkaloid composition. The aim of the research was to evaluate the antimicrobial potential of extracts from wild plants and in vitro cultures, as well as seven major individual alkaloids. Plant material derived from different natural habitats and in vitro cultures was used for the phytochemical analysis and antimicrobial tests. The composition of alkaloids was analyzed using chromatographic techniques (HPLC with DAD detection). The results have shown that roots contained higher number and amounts of alkaloids in comparison to aerial parts. All tested plant extracts manifested antimicrobial activity, related to different chemical structures of the alkaloids. Root extract used at 31.25-62.5 mg/L strongly reduced bacterial biomass. From the seven individually tested alkaloids, chelerythrine was the most effective against P. aeruginosa (MIC at 1.9 mg/L), while sanguinarine against S. aureus (MIC at 1.9 mg/L). Strong antifungal activity was observed against C. albicans when chelerythrine, chelidonine, and aerial parts extract were used. The experiments with plant extracts, individually tested alkaloids, and variable combinations of the latter allowed for a deeper insight into the potential mechanisms affecting the activity of this group of compounds.


Assuntos
Alcaloides/farmacologia , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Chelidonium , Isoquinolinas/farmacologia , Animais , Candida/efeitos dos fármacos , Candida/crescimento & desenvolvimento , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/crescimento & desenvolvimento , Camundongos , Componentes Aéreos da Planta , Raízes de Plantas , Staphylococcus aureus/crescimento & desenvolvimento
5.
PLoS One ; 14(6): e0217769, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31163049

RESUMO

Biofilm-related infections of bones pose a significant therapeutic issue. In this article we present in vitro results of the efficacy of gentamicin released from a collagen sponge carrier against Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella pneumoniae biofilms preformed on hydroxyapatite surface. The results indicate that high local concentrations of gentamicin released from a sponge eradicate the biofilm formed not only by gentamicin-sensitive strains but, to some extent, also by those that display a resistance pattern in routine diagnostics. The data presented in this paper is of high clinical translational value and may find application in the treatment of bone infections.


Assuntos
Biofilmes/efeitos dos fármacos , Colágeno/química , Liberação Controlada de Fármacos , Durapatita/química , Gentamicinas/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/ultraestrutura , Propriedades de Superfície
6.
J Pharm Biomed Anal ; 137: 13-22, 2017 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-28088662

RESUMO

Chronic leg ulceration is a disease usually associated with other comorbidities, and significantly reduces patient quality of life. Infected leg ulcers can lead to limb-threatening sequelae or mortality. Leg ulcerations are colonized by a number of microbes that are able to cause life-threating infections in susceptible patients. Wound exudate is a body fluid that collects metabolites from patient eukaryotic cells and from prokaryotic bacterial communities inhabiting the wound. This study aimed at identification of metabolites in exudates collected from chronic leg ulcers, and correlation of this metabolome with patient comorbidities and microbiological status of the wound. By means of NMR spectroscopy we detected 42 metabolites of microbial or patient origin. The metabolites that were in abundance in exudates analyzed were lactate, lysine, and leucine. Metabolites were associated with the presence of neutrophils in wounds and destruction of high quantities of microbes, but also with hypoxia typical for venous insufficiency. The combination of nuclear magnetic resonance spectroscopy technique and partial least squares discriminant analysis allowed us to further discriminate groups of metabolites with regards to potential clinical meaning. For example, to discriminate between S.aureus versus all other isolated microbial species, or between patients suffering from type I or II diabetes versus patients without diabetes. Therefore, wound exudate seems to be highly applicable material for discriminant analysis performed with the use of NMR technique to provide for rapid metabolomics of chronic wound status.


Assuntos
Líquidos Corporais/metabolismo , Complicações do Diabetes/metabolismo , Perna (Membro)/patologia , Metaboloma/fisiologia , Cicatrização/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença Crônica , Complicações do Diabetes/patologia , Diabetes Mellitus/metabolismo , Feminino , Humanos , Espectroscopia de Ressonância Magnética/métodos , Masculino , Metabolômica/métodos , Pessoa de Meia-Idade
7.
PLoS One ; 12(1): e0169565, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28076372

RESUMO

Bone infections are a significant public health burden associated with morbidity and mortality in patients. Microbial biofilm pathogens are the causative agents in chronic osteomyelitis. Research on the pathogenesis of osteomyelitis has focused on indirect bone destruction by host immune cells and cytokines secondary to microbial insult. Direct bone resorption by biofilm pathogens has not yet been seriously considered. In this study, common osteomyelitis pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Streptococcus mutans) were grown as biofilms in multiple in vitro and ex vivo experiments to analyze quantitative and qualitative aspects of bone destruction during infection. Pathogens were grown as single or mixed species biofilms on the following substrates: hydroxyapatite, rat jawbone, or polystyrene wells, and in various media. Biofilm growth was evaluated by scanning electron microscopy and pH levels were monitored over time. Histomorphologic and quantitative effects of biofilms on tested substrates were analyzed by microcomputed tomography and quantitative cultures. All tested biofilms demonstrated significant damage to bone. Scanning electron microscopy indicated that all strains formed mature biofilms within 7 days on all substrate surfaces regardless of media. Experimental conditions impacted pH levels, although this had no impact on biofilm growth or bone destruction. Presence of biofilm led to bone dissolution with a decrease of total volume by 20.17±2.93% upon microcomputed tomography analysis, which was statistically significant as compared to controls (p <0.05, ANOVA). Quantitative cultures indicated that media and substrate did not impact biofilm formation (Kruskall-Wallis test, post-hoc Dunne's test; p <0.05). Overall, these results indicate that biofilms associated with osteomyelitis have the ability to directly resorb bone. These findings should lead to a more complete understanding of the etiopathogenesis of osteomyelitis, where direct bone resorption by biofilm is considered in addition to the well-known osteoclastic and host cell destruction of bone.


Assuntos
Biofilmes/crescimento & desenvolvimento , Arcada Osseodentária/microbiologia , Osteoblastos/microbiologia , Osteomielite/microbiologia , Animais , Candida albicans/efeitos dos fármacos , Candida albicans/patogenicidade , Candida albicans/fisiologia , Durapatita/farmacologia , Arcada Osseodentária/patologia , Masculino , Osteoblastos/ultraestrutura , Poliestirenos/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/patogenicidade , Pseudomonas aeruginosa/fisiologia , Ratos , Ratos Wistar , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/patogenicidade , Staphylococcus aureus/fisiologia
8.
Carbohydr Polym ; 157: 371-379, 2017 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-27987940

RESUMO

The study aimed at evaluation of various types of alkali rinsing with regard to their efficacy in terms of removal, not only of bacteria but also bacterial metabolites, from cellulose matrices formed by three Komagataeibacter xylinus strains. Moreover, we tested the type of alkali rinsing on membrane cytotoxicity in vitro in fibroblast and osteoblast cells and we compared matrices' ability to induce oxidative stress in macrophages. We identified 11 metabolites of bacterial origin that remained in cellulose after rinsing. Moreover, our results indicated that the type of alkali rinsing should be adjusted to specific K. xylinus strains that are used as cellulose producers to obtain safe biomaterials in the context of low cytotoxicity and macrophage induction. The findings have translational importance and may be of direct significance to cellulose dressing manufacturers.


Assuntos
Materiais Biocompatíveis , Celulose/química , Gluconacetobacter xylinus/química , Álcalis , Animais , Bandagens , Linhagem Celular , Fibroblastos/efeitos dos fármacos , Humanos , Macrófagos/efeitos dos fármacos , Camundongos , Osteoblastos/efeitos dos fármacos
9.
Acta Bioeng Biomech ; 18(1): 115-20, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27150429

RESUMO

In our previous work we reported the impact of hydrofluoric and nitric acid used for chemical polishing of Ti-6Al-7Nb scaffolds on decrease of the number of Staphylococcus aureus biofilm forming cells. Herein, we tested impact of the aforementioned substances on biofilm of Gram-negative microorganism, Pseudomonas aeruginosa, dangerous pathogen responsible for plethora of implant-related infections. The Ti-6Al-7Nb scaffolds were manufactured using Selective Laser Melting method. Scaffolds were subjected to chemical polishing using a mixture of nitric acid and fluoride or left intact (control group). Pseudomonal biofilm was allowed to form on scaffolds for 24 hours and was removed by mechanical vortex shaking. The number of pseudomonal cells was estimated by means of quantitative culture and Scanning Electron Microscopy. The presence of nitric acid and fluoride on scaffold surfaces was assessed by means of IR and rentgen spetorscopy. Quantitative data were analysed using the Mann-Whitney test (P ≤ 0.05). Our results indicate that application of chemical polishing correlates with significant drop of biofilm-forming pseudomonal cells on the manufactured Ti-6Al-7Nb scaffolds ( p = 0.0133, Mann-Whitney test) compared to the number of biofilm-forming cells on non-polished scaffolds. As X-ray photoelectron spectroscopy revealed the presence of fluoride and nitrogen on the surface of scaffold, we speculate that drop of biofilm forming cells may be caused by biofilm-supressing activity of these two elements.


Assuntos
Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Lasers , Teste de Materiais/métodos , Pseudomonas aeruginosa/fisiologia , Titânio/farmacologia , Contagem de Colônia Microbiana , Flúor/análise , Nitrogênio/análise , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/ultraestrutura
10.
J Craniomaxillofac Surg ; 43(6): 863-9, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26027867

RESUMO

BACKGROUND: Because of the suspicion that bisphosphonates enhance bacterial colonization, this study evaluated adhesion and biofilm formation by Streptococcus mutans 25175, Staphylococcus aureus 6538, and Pseudomonas aeruginosa 14454 reference strains on hydroxyapatite coated with clodronate, pamidronate, or zoledronate. MATERIAL AND METHODS: Bacterial strains were cultured on bisphosphonate-coated and noncoated hydroxyapatite discs. After incubation, nonadhered bacteria were removed by centrifugation. Biofilm formation was confirmed by scanning electron microscopy. Bacterial colonization was estimated using quantitative cultures compared by means with Kruskal-Wallis and post-hoc Student-Newman-Keuls tests. Modeling of the interactions between bisphosphonates and hydroxyapatite was performed using the Density Functional Theory method. RESULTS: Bacterial colonization of the hydroxyapatite discs was significantly higher for all tested strains in the presence of bisphosphonates vs. CONTROLS: Adherence in the presence of pamidronate was higher than with other bisphosphonates. Density Functional Theory analysis showed that the protonated amine group of pamidronate, which are not present in clodronate or zoledronate, forms two additional hydrogen bonds with hydroxyapatite. Moreover, the reactive cationic amino group of pamidronate may attract bacteria by direct electrostatic interaction. CONCLUSION: Increased bacterial adhesion and biofilm formation can promote osteomyelitis, cause failure of dental implants or bisphosphonate-coated joint prostheses, and complicate bone surgery in patients on bisphosphonates.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Materiais Biocompatíveis/química , Biofilmes/efeitos dos fármacos , Conservadores da Densidade Óssea/farmacologia , Difosfonatos/farmacologia , Durapatita/química , Técnicas Bacteriológicas , Osso e Ossos , Ácido Clodrônico/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Humanos , Ligação de Hidrogênio/efeitos dos fármacos , Imidazóis/farmacologia , Microscopia Eletrônica de Varredura , Pamidronato , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Eletricidade Estática , Streptococcus mutans/efeitos dos fármacos , Microtomografia por Raio-X/métodos , Ácido Zoledrônico
11.
Wound Repair Regen ; 23(4): 525-30, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25899420

RESUMO

The purpose of this study was to compare the efficacy of dressings containing octenidine vs. dressings containing silver in the wound healing in the course of a chronic venous disease. There were two groups of 40 patients who met the inclusion criteria and who did not meet the exclusion criteria. The patients were randomly assigned into the groups (envelope method). The first, "O group" was treated with octenidine-based dressings. The second, "S group" was treated with silver dressings. The study lasted for 56 days. All patients in the research were treated with medical compression stockings with cotton understockings. Microbiological eradication was observed on the 28th day of the study among 33% of patients in the treatment group vs. 6% in control group. On the 56th day of the treatment, these percentages equalled 72% and 35%. The rate of healing was faster in the 0 group than in the S group. In the wounds <10 cm(2) it was faster by 1.35 cm(2)/week and in wounds >10 cm(2) it equalled 3.44 cm(2). The reduction of pain level was 37.5% higher in the O group, in contrast with the S group. One change of a dressing in the O group led to a 0.06 cm(2) greater wound size reduction and in the case of wounds >10 cm(2) to 0.29 cm(2) reduction compared with the S group. The presented results indicate that the efficacy of dressings containing octenidine is higher compared to silver dressings.


Assuntos
Úlcera da Perna/tratamento farmacológico , Piridinas/administração & dosagem , Prata/administração & dosagem , Meias de Compressão , Cicatrização/efeitos dos fármacos , Infecção dos Ferimentos/tratamento farmacológico , Administração Tópica , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anti-Infecciosos Locais/administração & dosagem , Doença Crônica , Feminino , Seguimentos , Humanos , Iminas , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Tempo , Resultado do Tratamento , Adulto Jovem
12.
J Oral Maxillofac Surg ; 73(3): 451-64, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25544303

RESUMO

PURPOSE: It is widely thought that inflammation and osteoclastogenesis result in hydroxyapatite (HA) resorption and sequestrum formation during osseous infections, and microbial biofilm pathogens induce the inflammatory destruction of HA. We hypothesized that biofilms associated with infectious bone disease can directly resorb HA in the absence of host inflammation or osteoclastogenesis. Therefore we developed an in vitro model to test this hypothesis. MATERIALS AND METHODS: Customized HA discs were manufactured as a substrate for growing clinically relevant biofilm pathogens. Single-species biofilms of Streptococcus mutans, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans and mixed-species biofilms of C albicans plus S mutans were incubated on HA discs for 72 hours to grow mature biofilms. Three different non-biofilm control groups also were established for testing. HA discs were then evaluated by means of scanning electron microscopy, micro-computed tomography metrotomography, x-ray spectroscopy, and confocal microscopy with planimetric analysis. In addition, quantitative cultures and pH assessment were performed. Analysis of variance was used to test for significance between treatment and control groups. RESULTS: All investigated biofilms were able to cause significant (P < .05) and morphologically characteristic alterations in HA structure as compared with controls. The highest number of alterations observed was caused by mixed biofilms of C albicans plus S mutans. S mutans biofilm incubated in medium with additional sucrose content was the most detrimental to HA surfaces among single-species biofilms. CONCLUSIONS: Our findings suggest that direct microbial resorption of bone is possible in addition to immune-mediated destruction, which has important translational implications for the pathogenesis of chronic bone infections and for targeted antimicrobial therapeutics.


Assuntos
Biofilmes , Durapatita/metabolismo , Carga Bacteriana , Técnicas Bacteriológicas , Biofilmes/crescimento & desenvolvimento , Candida albicans/crescimento & desenvolvimento , Candida albicans/metabolismo , Técnicas de Cocultura , Contagem de Colônia Microbiana , Meios de Cultura , Placa Dentária/metabolismo , Placa Dentária/microbiologia , Durapatita/química , Humanos , Concentração de Íons de Hidrogênio , Imageamento Tridimensional , Microscopia Confocal , Microscopia Eletrônica de Varredura , Micologia/métodos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/metabolismo , Espectrometria por Raios X , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/metabolismo , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/metabolismo , Sacarose/metabolismo , Tomografia , Microtomografia por Raio-X
13.
Int Wound J ; 11(6): 730-4, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23445335

RESUMO

Increasing data suggesting that microorganisms in the biofilm form are among the leading agents of persistent infections of chronic wounds require the development of new approaches to treatment. The aim of this article was to compare the efficacy of three commonly used antiseptics using a biofilm-oriented approach. Biofilm-oriented antiseptics test (BOAT), the innovative method, allows to estimate, in a quick and reliable manner, the in vitro activity of working solutions of antiseptics in real contact times against bacteria in the biofilm form and to use the results in the selection of an appropriate antiseptic to treat local infections in the clinical practice.


Assuntos
Anti-Infecciosos Locais/farmacologia , Biofilmes/efeitos dos fármacos , Etacridina/farmacologia , Povidona-Iodo/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Piridinas/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Iminas , Pseudomonas aeruginosa/fisiologia , Staphylococcus aureus/fisiologia
14.
Acta Bioeng Biomech ; 15(1): 69-76, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23957680

RESUMO

The Gram-positive coccus, Staphylococcus aureus, is the leading etiologic agent of limb and life-threatening biofilm-related infections in the patients following the orthopaedic implantations. The aim of the present paper is to estimate the ability of S. aureus to form biofilm on titanium alloy (Ti-6Al-7Nb) scaffolds produced by Selective Laser Melting (SLM) and subjected to the different types of surface modifications, including ultrasonic cleaning and chemical polishing. The results obtained indicate significantly the decreased ability of S.aureus to form biofilm on the surface of scaffolds subjected to the chemical polishing in comparison to the scaffolds cleaned ultrasonically. The data provided can be useful for future applications of the SLM technology in production of Ti-6Al-7Nb medical implants.


Assuntos
Biofilmes/crescimento & desenvolvimento , Lasers , Teste de Materiais/métodos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Alicerces Teciduais/química , Titânio/farmacologia , Biofilmes/efeitos dos fármacos , Staphylococcus aureus/citologia , Staphylococcus aureus/ultraestrutura , Propriedades de Superfície , Microtomografia por Raio-X
15.
J Oral Maxillofac Surg ; 71(6): 1010-6, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23489958

RESUMO

PURPOSE: Bacterial colonization of the denuded bone in bisphosphonate-related osteonecrosis of the jaw suggests that bisphosphonates increase bacterial adhesion and biofilm formation. This study evaluated the adhesion of gram-positive and gram-negative bacteria on hydroxyapatite coated with pamidronate, one of the most potent bisphosphonates. MATERIALS AND METHODS: Twenty-five Staphylococcus aureus and 25 Pseudomonas aeruginosa strains were cultured on pamidronate-coated and uncoated hydroxyapatite discs. After incubation, nonadherent bacteria were removed by rinsing and centrifugation. Formation of a biofilm was confirmed by confocal laser 3-dimensional and scanning electron microscopy. The number of bacterial colonies was counted using quantitative cultures and mean numbers were compared using the Mann-Whitney rank sum test (statistical significance defined as P ≤ .05). The Hartree-Fock method was used for the calculation of electron interactions between hydroxyapatite ions and pamidronate. RESULTS: Fold increases in the number of colonies formed by S aureus and P aeruginosa in the presence of pamidronate compared with controls were 7.19 ± 4.127 and 2.87 ± 0.622, respectively. Hartree-Fock analysis showed that the reactive NH3(+) group of pamidronate may act as a steric factor, facilitating anchoring of bacteria to the hydroxyapatite surface. Alternatively, the NH3(+) group may attract bacteria by direct electrostatic interaction. CONCLUSIONS: Increased bacterial adhesion in the presence of bisphosphonates can promote osteomyelitis in patients with bisphosphonate-related osteonecrosis of the jaw. There may be increased infection rates when bisphosphonates are used for stabilization of prostheses in joint arthroplasty and in osteotomies and open fractures in patients treated with bisphosphonates.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/microbiologia , Conservadores da Densidade Óssea/farmacologia , Difosfonatos/farmacologia , Durapatita , Biofilmes/efeitos dos fármacos , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/patologia , Contagem de Colônia Microbiana , Tomografia com Microscopia Eletrônica , Humanos , Modelos Moleculares , Pamidronato , Pseudomonas aeruginosa/fisiologia , Staphylococcus aureus/fisiologia , Estatísticas não Paramétricas , Microtomografia por Raio-X
16.
Acta Biochim Pol ; 60(4): 701-6, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24432320

RESUMO

Staphylococcus aureus is responsible for many types of infections related to biofilm presence. As the early diagnostics remains the best option for prevention of biofilm infections, the aim of the work presented was to search for differences in metabolite patterns of S. aureus ATCC6538 biofilm vs. free-swimming S. aureus planktonic forms. For this purpose, Nuclear Magnetic Resonance (NMR) spectroscopy was applied. Data obtained were supported by means of Scanning Electron Microscopy, quantitative cultures and X-ray computed microtomography. Metabolic trends accompanying S. aureus biofilm formation were found using Principal Component Analysis (PCA). Levels of isoleucine, alanine and 2,3-butanediol were significantly higher in biofilm than in planktonic forms, whereas level of osmoprotectant glycine-betaine was significantly higher in planktonic forms of S. aureus. Results obtained may find future application in clinical diagnostics of S. aureus biofilm-related infections.


Assuntos
Biofilmes/crescimento & desenvolvimento , Metaboloma , Infecções Estafilocócicas/diagnóstico , Staphylococcus aureus/metabolismo , Alanina/isolamento & purificação , Alanina/metabolismo , Biomarcadores/metabolismo , Butileno Glicóis/isolamento & purificação , Butileno Glicóis/metabolismo , Hidrogênio , Isoleucina/isolamento & purificação , Isoleucina/metabolismo , Espectroscopia de Ressonância Magnética , Radioisótopos , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/isolamento & purificação , Tomografia Computadorizada por Raios X
17.
Pol J Microbiol ; 61(3): 191-197, 2012 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-29334048

RESUMO

Roche's xCelligence impedance-measuring instrument is one of a few commercially available systems of such type. According to the best knowledge of authors, instrument was tested so far only for eukaryotic cell research. The aim of this work was to estimate xCELLigence suitability for the microbiological tests, including (i) measurement of morphological changes in eukaryotic cells as a result of bacterial toxin activity, (ii) measurement of bacterial biofilm formation and (iii) impact of antiseptics on the biofilm structure. To test the infuence of bacterial LT enterotoxin on eukaryotic cell lines, Chinese Hamster Ovary (CHO) cell line and reference strain Escherichia coli ATTC 35401 were used. To investigate Roche's instrument ability to measure biofilm formation and impact of antiseptics on its development, Staphylococcus aureus ATTC6538 reference strain was used. The data generated during the experiments indicate excellent ability of xCelligence instrument to detect cytopathic effect caused by bacterial LT endotoxin and to detect staphylococcal biofilm formation. However, interpretation of the results obtained during real-time measurement of antiseptic's bactericidal activity against staphylococcal biofilm, caused many difficulties. xCelligence instrument can be used for real-time monitoring of morphological changes in CHO cells treated with bacterial LT enterotoxin and for real-time measurement of staphylococcal biofilm formation in vitro. Further investigation is necessary to confirm suitability of system to analyze antiseptic's antimicrobial activity against biofilm in vitro.

18.
Otolaryngol Pol ; 59(4): 517-22, 2005.
Artigo em Polonês | MEDLINE | ID: mdl-16273854

RESUMO

UNLABELLED: 40 patients with exacerbation of a chronic maxillary sinusitis were examined. Sinus puncture was performed (sinoject) in all of the patients. Before the treatment (500 mg cefprozil orally twice a day, the recommended duration of therapy was ten days) and on the fifth day, the microorganisms from sinus were isolated (the bacteriological culture, antibiogram and MIC were determined). On the second and the fifth day, blood cefprozil level and the presence of cefprozil in the washings from maxillary sinuses were investigated. RESULTS: 17 pathogens were isolated from maxillary sinuses, only two of them were anaerobic. The bacterial strains: Staphylococcus (55%), E.coli (15%), Klebsiella (10%) and 20% of others, were found. Four of them (23.5%) were resistant to cefprozil. In all patients cefprozil was identified in maxillary sinuses (0.87-2.52 ug/ml). The value of MIC were from 0.094 to 2.0 ug/ml. A satisfactory clinical response was observed in 92.5% persons but the eradication of pathogens was obtained in 70%. The adverse clinical events (diarrhea) were observed only in one patient (2.5%). CONCLUSIONS: cefprozil well penetrates into inflammable mucous membrane of maxillary sinuses. Cefprozil obtains efficient bactericidal concentration in relation to sensitive bacteria on the fifth day of therapy. It is also well tolerated by the patients.


Assuntos
Antibacterianos/farmacologia , Cefalosporinas/farmacologia , Sinusite Maxilar/tratamento farmacológico , Sinusite Maxilar/microbiologia , Adolescente , Adulto , Doença Crônica , Infecções por Escherichia coli/tratamento farmacológico , Feminino , Humanos , Infecções por Klebsiella/tratamento farmacológico , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Infecções Estafilocócicas/tratamento farmacológico , Resultado do Tratamento , Cefprozil
19.
Med Dosw Mikrobiol ; 55(2): 135-46, 2003.
Artigo em Polonês | MEDLINE | ID: mdl-14577193

RESUMO

Colonisation and remaining of microorganism on mucus membrane of microorganism is tightly connected with adhesion mechanisms and determine the first step of physiological settlement of the organism or the first stage of clinically demonstrated infection. In Klebsiella rods there are known three types of fimbrial adhesins (type 1, 3 and KPF-28) and non-fimbrial adhesin CF29K. It is stated that Klebsiella strains adhesions are responsible for their adherence to the epithelial cells of both respiratory and urinary tracts and to intestine epithelium. The in vitro research affirmed Klebsiella rods adherence to protein matrix. The aim of our work was the establishment of character, receptor specificity and the appearance frequency of P-like called adhesin. The frequency of expression of P-like adhesin was estimated among 380 isolated from the patients strains on the basis of agglutinating methods. The amorphic character of P-like adhesin was proved using electron microscopy method. The isolation and purification of P-like protein with a help of affinity chromatography enabled to estimate the receptor specificity of the adhesin. The receptor specificity was established as similar to E.coli PapG adhesin.


Assuntos
Adesinas Bacterianas/fisiologia , Klebsiella/patogenicidade , Adesinas Bacterianas/isolamento & purificação , Aderência Bacteriana , Humanos , Técnicas In Vitro , Klebsiella/isolamento & purificação , Microscopia Eletrônica , Especificidade da Espécie
20.
Med Dosw Mikrobiol ; 55(4): 333-42, 2003.
Artigo em Polonês | MEDLINE | ID: mdl-15103992

RESUMO

Gram negative Klebsiella bacilli present many pathogenic properties, which determine their ability to survive and rapid spreading in hospital environment. There are many factors responsible for the pathogenicity of Klebsiella strains: capsule, fimbriae, nonfimbrial adhesins, lipopolysaccharide of the cell wall and extracellular secreted exotoxins. Klebsiella strains are etiological agents of different nosocomial infections but also colonized gastrointestinal and respiratory tracts. The aim of our work were adhesive properties and antibiotic resistance of Klebsiella strains isolated from stool of hospitalized children, according to source of potential nosocomial infections--100 Klebsiella strains from Wroclaw and 76 strains from Opole, isolated in cases of diarrhea. The resistance of this strains to different group of antibiotics, the expression of ESBL enzymes, the activity in hemagglutination and their ability to adherence to different cell lines were tested. The highest resistance of all strains to aminopenicillins was observed. The production of ESBL was highest in strains from Opole (51% strains) then in Wroclaw (9%). In both hospital units, ESBL+ strains were resistant to aminoglicosides and cotrimoxazol but sensitive to ciprofloxacine. Using hemagglutination method the types of fimbriae were defined. Above 90% investigated Klebsiella strains showed the presence of fimbriae (in Wroclaw more strains simultaneously expressed fimbriae type 1 and 3, in Opole mainly fimbriae type 3). Over 70% strains demonstrated the high level of adherence to cell lines. Only several strains showed the low level or the lack of adhesion. These results suggested that among Klebsiella strains in gastrointestinal tract were presented multiresistant strains with high ability to adherence, which may be potential source of nosocomial infections.


Assuntos
Aderência Bacteriana , Farmacorresistência Bacteriana , Intestinos/microbiologia , Infecções por Klebsiella/microbiologia , Klebsiella/patogenicidade , Criança , Infecções por Enterobacteriaceae/microbiologia , Humanos , Klebsiella/efeitos dos fármacos , Klebsiella/isolamento & purificação
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