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1.
Int J Mol Sci ; 22(3)2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33530493

RESUMO

The impact of the Gram-negative bacterium Escherichia coli (E. coli) on the microbiomic and pathogenic phenomena occurring in humans and other warm-blooded animals is relatively well-recognized. At the same time, there are scant data concerning the role of E. coli strains in the health and disease of cold-blooded animals. It is presently known that reptiles are common asymptomatic carriers of another human pathogen, Salmonella, which, when transferred to humans, may cause a disease referred to as reptile-associated salmonellosis (RAS). We therefore hypothesized that reptiles may also be carriers of specific E. coli strains (reptilian Escherichia coli, RepEC) which may differ in their genetic composition from the human uropathogenic strain (UPEC) and avian pathogenic E. coli (APEC). Therefore, we isolated RepECs (n = 24) from reptile feces and compared isolated strains' pathogenic potentials and phylogenic relations with the aforementioned UPEC (n = 24) and APEC (n = 24) strains. To this end, we conducted an array of molecular analyses, including determination of the phylogenetic groups of E. coli, virulence genotyping, Pulsed-Field Gel Electrophoresis-Restriction Analysis (RA-PFGE) and genetic population structure analysis using Multi-Locus Sequence Typing (MLST). The majority of the tested RepEC strains belonged to nonpathogenic phylogroups, with an important exception of one strain, which belonged to the pathogenic group B2, typical of extraintestinal pathogenic E. coli. This strain was part of the globally disseminated ST131 lineage. Unlike RepEC strains and in line with previous studies, a high percentage of UPEC strains belonged to the phylogroup B2, and the percentage distribution of phylogroups among the tested APEC strains was relatively homogenous, with most coming from the following nonpathogenic groups: C, A and B1. The RA-PFGE displayed a high genetic diversity among all the tested E. coli groups. In the case of RepEC strains, the frequency of occurrence of virulence genes (VGs) was lower than in the UPEC and APEC strains. The presented study is one of the first attempting to compare the phylogenetic structures of E. coli populations isolated from three groups of vertebrates: reptiles, birds and mammals (humans).


Assuntos
Doenças dos Animais/microbiologia , Infecções por Escherichia coli/veterinária , Filogenia , Répteis/microbiologia , Escherichia coli Uropatogênica/classificação , Escherichia coli Uropatogênica/genética , Animais , Proteínas de Escherichia coli/genética , Especificidade de Hospedeiro , Humanos , Tipagem de Sequências Multilocus , Doenças das Aves Domésticas/microbiologia , Virulência/genética , Fatores de Virulência/genética
2.
Int J Mol Sci ; 23(1)2021 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-35008720

RESUMO

The aim of this study was to assess the beneficial inhibitory effect of silver nanoparticles immobilized on SiO2 or TiO2 on biofilm formation by Pseudomonas aeruginosa-one of the most dangerous pathogens isolated from urine and bronchoalveolar lavage fluid of patients hospitalized in intensive care units. Pure and silver doped nanoparticles of SiO2 and TiO2 were prepared using a novel modified sol-gel method. Ten clinical strains of P. aeruginosa and the reference PAO1 strain were used. The minimal inhibitory concentration (MIC) was determined by the broth microdilution method. The minimal biofilm inhibitory concentration (MBIC) and biofilm formation were assessed by colorimetric assay. Bacterial enumeration was used to assess the viability of bacteria in the biofilm. Silver nanoparticles immobilized on the SiO2 and TiO2 indicated high antibacterial efficacy against P. aeruginosa planktonic and biofilm cultures. TiO2/Ag0 showed a better bactericidal effect than SiO2/Ag0. Our results indicate that the inorganic compounds (SiO2, TiO2) after nanotechnological modification may be successfully used as antibacterial agents against multidrug-resistant P. aeruginosa strains.

3.
Pathogens ; 9(8)2020 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-32784631

RESUMO

Salmonella enterica ser. Enteritidis (S. enterica ser. Enteritidis) is the most frequently detected serovar in human salmonellosis, and its ability to produce a biofilm and the risk of transmission from animals and food of animal origin to humans are significant. The main aim of the present work was to compare S. enterica ser. Enteritidis strains isolated from poultry and human feces in terms of resistance profiles, prevalence of selected resistance genes, and their potential for biofilm formation, by assessing their biofilm growth intensity, the prevalence and expression of selected genes associated with this phenomenon, and the correlation between increased antimicrobial resistance and biofilm formation ability of the two tested groups of S. enterica ser. Enteritidis. This study showed a difference in antimicrobial resistance (minimal inhibitory concentration value) between S. enterica ser. Enteritidis groups; however, the majority of multidrug-resistant (MDR) strains were isolated from poultry (environmental samples from chicken broilers, turkey broilers, and laying hens). Differences in the prevalence of resistance genes were observed; the most common gene among poultry strains was floR, and that among strains from humans was blaTEM. S. enterica ser. Enteritidis strains isolated from poultry under the tested incubation conditions exhibited better biofilm growth than strains isolated from humans. A higher level of gene expression associated with the production of cellulose was only detected in the S48 strain isolated from poultry. On the other hand, increased expression of genes associated with quorum sensing was observed in two strains isolated from poultry farms and one strain isolated from human feces.

4.
Int J Nanomedicine ; 15: 199-213, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32021174

RESUMO

Purpose: Resistance to antibiotics is a major problem of public health. One of the alternative therapies is silver - more and more popular because of nanotechnology development and new possibilities of usage. As a component of colloid, powder, cream, bandages, etc., nanosilver is often recommended to treat the multidrug-resistant pathogens and we can observe its overuse also outside of the clinic where different physicochemical forms of silver nanoformulations (e.g. size, shape, compounds, surface area) are introduced. In this research, we described the consequences of long-term bacteria exposure to silver nanoformulations with different physicochemical properties, including changes in genome and changes of bacterial sensitivity to silver nanoformulations and/or antibiotics. Moreover, the prevalence of exogenous resistance to silver among multidrug-resistant bacteria was determined. Materials and Methods: Gram-negative and Gram-positive bacteria strains are described as sensitive and multidrug-resistant strains. The sensitivity of the tested bacterial strains to antibiotics was carried out with disc diffusion methods. The sensitivity of bacteria to silver nanoformulations and development of bacterial resistance to silver nanoformulations has been verified via determination of the minimal inhibitory concentrations. The presence of sil genes was verified via PCR reaction and DNA electrophoresis. The genomic and phenotypic changes have been verified via genome sequencing and bioinformatics analysis. Results: Bacteria after long-term exposure to silver nanoformulations may change their sensitivity to silver forms and/or antibiotics, depending on the physicochemical properties of silver nanoformulations, resulting from phenotypic or genetic changes in the bacterial cell. Finally, adaptants and mutants may become more sensitive or resistant to some antibiotics than wild types. Conclusion: Application of silver nanoformulations in the case of multiple resistance or multidrug-resistant bacterial infection can enhance or decrease their resistance to antibiotics. The usage of nanosilver in a clinic and outside of the clinic should be determined and should be under strong control. Moreover, each silver nanomaterial should be considered as a separate agent with a potential different mode of antibacterial action.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Nanoestruturas/química , Prata/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla/genética , Bactérias Gram-Negativas/genética , Bactérias Gram-Positivas/genética , Testes de Sensibilidade Microbiana , Prata/química
5.
Mater Sci Eng C Mater Biol Appl ; 106: 110295, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31753350

RESUMO

Hexagonal nanocrystalline powders of the non-doped Ca10(PO4)6(OH)2 as well as activated with Ag+ and Eu3+ ions were synthesized by using different wet chemistry methods. Moreover, the obtained hydroxyapatite was loaded with Ag0, as well as nitroimidazole antimicrobials: metronidazole and tinidazole. The structural properties of the products were analyzed by X-ray diffraction (XRD), scanning (SEM) and transmission (TEM) electron microscopy as well as infrared (IR) and Raman spectroscopy. The photoluminescence properties of the Eu3+ and Ag+ co-doped Ca10(PO4)6(OH)2 were characterized via the PL emission, excitation spectra and the luminescence decay curve. The antimicrobial activity of the obtained materials against Prevotella bivia and Parabacteroides distasonis was studied. The cytotoxicity assessment was carried out on the human osteosarcoma cell line (U2OS) as well as human red blood cells (RBC). The choice of the in vitro model was based on the fact that U2OS is a cancer cell line derived from bone tissue which is rich in apatites that play a pivotal role in the extracellular matrix formation. RBCs are the most abundant blood cells and they are used as a cell model in the study of biocompatibility of new prepared biocompounds with potential medical applications. The obtained multifunctional materials do not exhibit the haemolytic activity, therefore, they could be used as a promising antimicrobial agent and for anaerobic bacteria.


Assuntos
Bacteroidetes/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Hidroxiapatitas/química , Nanocompostos/química , Prevotella/efeitos dos fármacos , Adsorção , Animais , Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/química , Sedimentação Sanguínea/efeitos dos fármacos , Bovinos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Európio/química , Hemólise/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Muramidase/química , Nanocompostos/toxicidade , Soroalbumina Bovina/química , Prata/química
6.
BMC Vet Res ; 15(1): 312, 2019 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-31477105

RESUMO

BACKGROUND: Salmonella is generally considered as a human pathogen causing typhoid fever and gastrointestinal infections called salmonellosis, with S. Enteritidis and S. Typhimurium strains as the main causative agents. Salmonella enterica strains have a wide host array including humans, birds, pigs, horses, dogs, cats, reptiles, amphibians and insects. Up to 90% of reptiles are the carriers of one or more serovars of Salmonella. Extraintestinal bacterial infections associated with reptiles pose serious health threat to humans. The import of exotic species of reptiles as pet animals to Europe correlates with the emergence of Salmonella serotypes, which not found previously in European countries. The presented study is a new report about Salmonella serotypes associated with exotic reptiles in Poland. The goal of this research was to examine the zoonotic potential of Salmonella strains isolated from reptiles by comparative analysis with S. Enteritidis strains occurring in human population and causing salmonellosis. RESULTS: The main findings of our work show that exotic reptiles are asymptomatic carriers of Salmonella serovars other than correlated with salmonellosis in humans (S. Enteritidis, S. Typhimurium). Among the isolated Salmonella strains we identified serovars that have not been reported earlier in Poland, for example belonging to subspecies diarizonae and salamae. Restriction analysis with Pulsed-field Gel Electrophoresis (PFGE), showed a great diversity among Salmonella strains isolated from reptiles. Almost all tested strains had distinct restriction patterns. While S. Enteritidis strains were quite homogeneous in term of phylogenetic relations. Most of the tested VGs were common for the two tested groups of Salmonella strains. CONCLUSIONS: The obtained results show that Salmonella strains isolated from reptiles share most of virulence genes with the S. Enteritidis strains and exhibit a greater phylogenetic diversity than the tested S. Enteritidis population.


Assuntos
Eletroforese em Gel de Campo Pulsado , Répteis/microbiologia , Infecções por Salmonella/microbiologia , Salmonella enterica/genética , Animais , Portador Sadio , Cromatografia Gasosa , DNA Bacteriano , Genótipo , Humanos , Salmonella enterica/patogenicidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem , Virulência , Zoonoses
7.
Microb Drug Resist ; 25(2): 277-286, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30698495

RESUMO

Enterococci are a natural component of the intestinal flora of many organisms, including humans and birds. As opportunistic pathogens, they can cause fatal infections of the urinary tract and endocarditis in humans, whereas in poultry symptoms are joint disease, sepsis, and falls in the first week of life. The study covered 107 Enterococcus strains-56 isolated from humans and 51 from turkeys. Among the isolates investigated Enterococcus faecalis was detected in 80.36% of human and 80.39% of turkey samples. Enterococcus faecium was identified in 8.93% of human and 17.65% of turkey strains. The highest percentage of the strains was resistant to tetracycline as follows: 48 (85.71%) and 48 (94.12%) of human and turkey strains, respectively. Resistance to erythromycin occurred in 37.50% of the human and in 76.47% of turkey strains, otherwise 27.10% of all strains showed resistance to ciprofloxacin. Our study revealed that 25% of human and 15.69% of turkey strains were resistant to vancomycin. Multidrug resistance showed in 32.14% and 43.14% of human and turkey strains, respectively. The tetracycline resistance gene, tetM, was detected in 82.24% of all strains analyzed, whereas the tetO gene was found in 53.57% of human but only in 7.84% of turkey strains. The vancomycin resistance gene (vanA) was detected in seven Enterococcus strains (six isolated from turkeys and one from humans). The ermB gene (resistance to macrolide) was detected in 55.14% of all isolates (42.86% of human and 68.63% of turkey strains), whereas the ermA gene was detected in 17.65% of turkey but only in 3.57% of human isolates. All the strains had the ability to form biofilms. A stronger biofilm was formed after 24-hour incubation by strains isolated from turkeys, whereas after 48 hours of incubation all examined strains produced strong biofilm.


Assuntos
Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Farmacorresistência Bacteriana Múltipla/genética , Enterococcus/efeitos dos fármacos , Enterococcus/genética , Perus/microbiologia , Animais , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/genética , Enterococcus faecium/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Polônia/epidemiologia
8.
Curr Med Chem ; 26(11): 1979-1993, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30207213

RESUMO

The environment exerts strong influence on microbes. Adaptation of microbes to changing conditions is a dynamic process regulated by complex networks. Pseudomonas aeruginosa is a life-threating, versatile opportunistic and multi drug resistant pathogen that provides a model to investigate adaptation mechanisms to environmental changes. The ability of P. aeruginosa to form biofilms and to modify virulence in response to environmental changes is coordinated by various mechanisms including two-component systems (TCS), and secondary messengers involved in quorum sensing (QS) and c-di-GMP networks (diguanylate cyclase systems, DGC). In this review, we focus on the role of c-di-GMP during biofilm formation. We describe TCS and QS signal cascades regulated by c-di-GMP in response to changes in the external environment. We present a complex signaling network dynamically changing during the transition of P. aeruginosa from the free-living to sessile mode of growth.


Assuntos
Biofilmes , GMP Cíclico/análogos & derivados , Pseudomonas aeruginosa/fisiologia , Percepção de Quorum/fisiologia , Transdução de Sinais/fisiologia , GMP Cíclico/metabolismo , Pseudomonas aeruginosa/crescimento & desenvolvimento
9.
BMC Microbiol ; 18(1): 86, 2018 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-30119648

RESUMO

BACKGROUND: Yersinia enterocolitica is widespread within the humans, pigs and wild boars. The low isolation rate of Y. enterocolitica from food or environmental and clinical samples may be caused by limited sensitivity of culture methods. The main goal of present study was identification of presumptive Y. enterocolitica isolates using MALDI TOF MS. The identification of isolates may be difficult due to variability of bacterial strains in terms of biochemical characteristics. This work emphasizes the necessity of use of multiple methods for zoonotic Y. enterocolitica identification. RESULTS: Identification of Y. enterocolitica isolates was based on MALDI TOF MS, and verified by VITEK® 2 Compact and PCR. There were no discrepancies in identification of all human' and pig' isolates using MALDI TOF MS and VITEK® 2 Compact. However three isolates from wild boars were not decisively confirmed as Y. enterocolitica. MALDI TOF MS has identified the wild boar' isolates designated as 3dz, 4dz, 8dz as Y. enterocolitica with a high score of matching with the reference spectra of MALDI Biotyper. In turn, VITEK® 2 Compact identified 3dz and 8dz as Y. kristensenii, and isolate 4dz as Y. enterocolitica. The PCR for Y. enterocolitica 16S rDNA for these three isolates was negative, but the 16S rDNA sequence analysis identified these isolates as Y. kristensenii (3dz, 4dz) and Y. pekkanenii (8dz). The wild boar' isolates 3dz, 4dz and 8dz could not be classified using biotyping. The main bioserotype present within pigs and human faeces was 4/O:3. It has been shown that Y. enterocolitica 1B/O:8 can be isolated from human faeces using ITC/CIN culturing. CONCLUSION: The results of our study indicate wild boars as a reservoir of new and atypical strains of Yersinia, for which protein and biochemical profiles are not included in the MALDI Biotyper or VITEK® 2 Compact databases. Pigs in the south-west Poland are the reservoir for pathogenic Y. enterocolitica strains. Four biochemical features included in VITEK® 2 Compact known to be common with Wauters scheme were shown to produce incompatible results, thus VITEK® 2 Compact cannot be applied in biotyping of Y. enterocolitica.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Sus scrofa/microbiologia , Suínos/microbiologia , Yersinia enterocolitica/isolamento & purificação , Animais , DNA Ribossômico , Reservatórios de Doenças/microbiologia , Fezes/microbiologia , Humanos , Polônia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , Análise de Sequência , Especificidade da Espécie , Yersinia/classificação , Yersinia/genética , Yersinia/isolamento & purificação , Yersinia enterocolitica/classificação , Yersinia enterocolitica/genética
10.
Microb Drug Resist ; 24(3): 314-322, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28628752

RESUMO

Campylobacter spp. is a major cause of foodborne diseases in humans, particularly when transmitted by the handling or consumption of undercooked poultry meat. Most Campylobacter infections are self-limiting, but antimicrobial treatment (e.g., fluoroquinolones and macrolides) is necessary in severe or prolonged cases. The indiscriminate use of these drugs, both in clinical medicine and animal production, has a major impact on public health. The aim of the present study was to identify Campylobacter strains, isolated from turkey and broilers, using both PCR and the matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) methods to reveal the accuracy of identification, as well to evaluate the antimicrobial and genetic resistance of the investigated strains. MALDI-TOF and PCR methods were used to show differences, if any, in the specificity of that test. In this study, MALDI-TOF mass spectrometry gave the same results as multiplex PCR, in all cases. The highest rate of resistance (i.e., 100% of turkey and broiler strains) was detected against ciprofloxacin, whereas 58.1% of turkey and 78.6% of broiler strains were resistant to tetracycline. Multidrug-resistant isolates were not found in the study. All ciprofloxacin-resistant strains had a mutation in the gyrA gene, at the Thr-86 position. The presence of the tetO gene was found in 71% of turkey and in 100% of broiler strains. All resistant to tetracycline strains included tetO gene. Additionally, in five turkey and three broiler strains, susceptible to tetracycline, tetO gene was present. These results indicate the high prevalence of Campylobacter strains, which are phenotypically and genetically resistant to fluoroquinolones and tetracycline.


Assuntos
Antibacterianos/farmacologia , Infecções por Campylobacter/veterinária , Campylobacter coli/genética , Campylobacter jejuni/genética , Farmacorresistência Bacteriana/genética , Doenças das Aves Domésticas/epidemiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Campylobacter coli/classificação , Campylobacter coli/efeitos dos fármacos , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/classificação , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/isolamento & purificação , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Galinhas/microbiologia , Ciprofloxacina/farmacologia , DNA Girase/genética , DNA Girase/metabolismo , Fazendas , Fezes/microbiologia , Expressão Gênica , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase Multiplex/veterinária , Polônia/epidemiologia , Aves Domésticas/microbiologia , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Tetraciclina/farmacologia , Perus/microbiologia
11.
Adv Clin Exp Med ; 26(3): 483-490, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28791824

RESUMO

BACKGROUND: Asiatic (AA) and ursolic (UA) acids are widely studied phytochemicals, but their antimicrobial properties are still poorly understood. Therefore our research has focused on their activity against uropathogenic Enterococcus faecalis strains. OBJECTIVES: The aim of this research was to determine the influence of AA and UA on the growth, cell morphology, virulence factors and biofilm formation by E. faecalis strains. MATERIAL AND METHODS: AA and UA were purchased from Sigma-Aldrich. E. faecalis strains were isolated from the urine samples of patients with urinary tract infections. The strains were checked for the presence of virulence genes using the PCR method. Their antimicrobial susceptibility was performed using the disc diffusion method. The MICs of triterpenes were determined using the broth microdilution method. The hydrophobicity of cells was established by salt aggregation test. Lipase and lecithinase activities were determined by using an agar medium containing egg yolk emulsion. DNase agar was used for the detection of DNase synthesis. Hemolytic activity was established using a sheep-blood agar. Todd-Hewitt agar medium containing gelatin was used for determination of gelatinase activity. The anti-biofilm activity of asiatic acid and ursolic acid was tested on polystyrene microtiter plates. It was examined using time-kill and biofilm assays. RESULTS: Reduction of growth and enzyme synthesis after exposure of E. faecalis to the acids was observed. None of the acids changed the hydrophobicity of bacteria. Stronger anti-biofilm activity was observed when the bacteria were incubated with AA. Thus, reduction of both the survival and the virulence factors will make bacteria less infectious. CONCLUSIONS: Based on the results obtained, we can assume that the triterpenes investigated should be considered natural components of a human diet rather than as antibacterial agents used on their own.


Assuntos
Antibacterianos/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Triterpenos Pentacíclicos/farmacologia , Biofilmes/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/metabolismo , Humanos , Lipase/metabolismo , Testes de Sensibilidade Microbiana/métodos , Fosfolipases/metabolismo , Triterpenos/farmacologia , Virulência/efeitos dos fármacos , Fatores de Virulência/metabolismo
12.
Int J Mol Sci ; 18(7)2017 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-28696348

RESUMO

A new emerging phenomenon is the association between the incorrect use of biocides in the process of disinfection in farms and the emergence of cross-resistance in Salmonella populations. Adaptation of the microorganisms to the sub-inhibitory concentrations of the disinfectants is not clear, but may result in an increase of sensitivity or resistance to antibiotics, depending on the biocide used and the challenged Salmonella serovar. Exposure of five Salmonella enterica subsp. enterica serovar Senftenberg (S. Senftenberg) strains to triamine-containing disinfectant did not result in variants with resistance to antibiotics, but has changed their susceptibility to normal human serum (NHS). Three biocide variants developed reduced sensitivity to NHS in comparison to the sensitive parental strains, while two isolates lost their resistance to serum. For S. Senftenberg, which exhibited the highest triamine tolerance (6 × MIC) and intrinsic sensitivity to 22.5% and 45% NHS, a downregulation of flagellin and enolase has been demonstrated, which might suggest a lower adhesion and virulence of the bacteria. This is the first report demonstrating the influence of biocide tolerance on NHS resistance. In conclusion, there was a potential in S. Senftenberg to adjust to the conditions, where the biocide containing triamine was present. However, the adaptation did not result in the increase of antibiotic resistance, but manifested in changes within outer membrane proteins' patterns. The strategy of bacterial membrane proteins' analysis provides an opportunity to adjust the ways of infection treatments, especially when it is connected to the life-threating bacteremia caused by Salmonella species.


Assuntos
Anti-Infecciosos/farmacologia , Proteínas de Bactérias/metabolismo , Desinfetantes/farmacologia , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/metabolismo , Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Humanos , Testes de Sensibilidade Microbiana
13.
PLoS One ; 11(10): e0164069, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27695090

RESUMO

Differential analysis of outer membrane composition of S. Enteritidis strains, resistant to 50% normal human serum (NHS) was performed in order to find factors influencing the resistance to higher concentrations of NHS. Ten S. Enteritidis clinical strains, resistant to 50% NHS, all producing very long lipopolysaccharide, were subjected to the challenge of 75% NHS. Five extreme strains: two resistant and three sensitive to 75% NHS, were chosen for the further analysis of outer membrane proteins composition. Substantial differences were found in the levels of particular outer membrane proteins between resistant and sensitive strains, i.e. outer membrane protease E (PgtE) was present mainly in resistant strains, while sensitive strains possessed a high level of flagellar hook-associated protein 2 (FliD) and significantly higher levels of outer membrane protein A (OmpA).


Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Soros Imunes/imunologia , Proteoma , Proteômica , Salmonella enteritidis/imunologia , Salmonella enteritidis/metabolismo , Complemento C3/imunologia , Feminino , Humanos , Lipopolissacarídeos/imunologia , Masculino , Viabilidade Microbiana/imunologia , Testes de Neutralização , Estabilidade Proteica , Proteômica/métodos , Temperatura
14.
Int J Food Sci Nutr ; 67(8): 1005-16, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27456160

RESUMO

Drinking of cranberry fruit juice and application of commercial preparations containing the cranberry extracts are recommended in the prevention and treatment of urinary tract infections (UTIs), especially in women with recurrent UTIs. Many studies focus on the activity of cranberries against uropathogenic Escherichia coli (E. coli) strains. However, the knowledge of the cranberry effect on Gram-positive Enterococcus faecalis (E. faecalis) is limited. Therefore, the aim of our study was to establish the activity of commercial concentrated cranberry extract on the growth, virulence factors and biofilm formation of E. faecalis strains isolated from urine. Minimal inhibitory concentrations (MICs) of cranberry extract were determined by the broth microdilution method. Disc diffusion method was used to determine antimicrobial susceptibility. The impact of cranberry extract on bacterial survival, hydrophobicity, synthesis of lipase, lecithinase, DNase, hemolysin, gelatinase and biofilm mass was determined. Results show that cranberry extract inhibits the growth, enzymatic activities of bacteria and limits biofilm formation. The antibacterial activities of the studied cranberry extract confirm that it could be successfully used in prevention of UTIs caused by E. faecalis.


Assuntos
Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/patogenicidade , Infecções por Bactérias Gram-Positivas/prevenção & controle , Infecções Urinárias/prevenção & controle , Vaccinium macrocarpon/química , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Enterococcus faecalis/fisiologia , Sucos de Frutas e Vegetais/análise , Genes Bacterianos/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Técnicas In Vitro , Extratos Vegetais/farmacologia , Infecções Urinárias/microbiologia , Fatores de Virulência/biossíntese , Fatores de Virulência/genética
15.
Curr Microbiol ; 72(5): 570-82, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26804795

RESUMO

In diagnostic microbiology as well as in microbiological research, the identification of a microorganism is a crucial and decisive stage. A broad choice of methods is available, based on both phenotypic and molecular properties of microbes. The aim of this study was to compare the application of phenotypic and molecular tools in bacterial identification on the example of Gram-negative intestine rod with an ambiguous phenotype. Different methods of identification procedure, which based on various properties of bacteria, were applied, e.g., microscopic observation of single-bacterial cells, macroscopic observation of bacterial colonies morphology, the automated system of microorganism identification (biochemical tests), the mass spectrometry method (analysis of bacterial proteome), and genetic analysis with PCR reactions. The obtained results revealed discrepancies in the identification of the tested bacterial strain with an atypical phenotype: mucous morphology of colonies, not characteristic for either E. coli and Citrobacter spp., mass spectrometry analysis of proteome initially assigned the tested strain to Citrobacter genus (C. freundii) and biochemical profiles pointed to Escherichia coli. A decisive method in the current study was genetic analysis with PCR reactions which identified conserved genetic sequences highly specific to E. coli species in the genome of the tested strain.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Bactérias Gram-Negativas/isolamento & purificação , Infecções por Bactérias Gram-Negativas/microbiologia , Reação em Cadeia da Polimerase/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Técnicas de Tipagem Bacteriana/instrumentação , Fezes/microbiologia , Bactérias Gram-Negativas/química , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/diagnóstico , Humanos , Fenótipo
16.
J Food Prot ; 73(7): 1325-7, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20615346

RESUMO

In this study, the molecular characteristics of food-derived oxacillin-resistant Staphylococcus aureus were determined. Eight borderline oxacillin-resistant strains with MICs of 2 to 4 microg/ml were identified from 132 S. aureus isolates of food origin. One of the two isolates with a MIC of 4 microg/ml was methicillin-resistant determinant (mecA) gene positive, and the other six with MICs of 2 microg/ml were mecA negative. The mecA-positive isolate was classified as sequence type (ST)228, staphylococcal protein A (spa) type t041, and carried the staphylococcal cassette chromosome mec type I element. Two borderline oxacillin-resistant strains were classified as spa t008 and ST8, and the remaining five as spa t164 and ST20. The mecA-positive strain and four borderline oxacillin-resistant strains were found enterotoxigenic. The enterotoxin genes detected in these strains included selp, egc1, and sed-sej-selr. The borderline-resistant S. aureus isolates from a manually handled product, i.e., minced pork, were shown genetically related to strains associated with human infections. This suggests that humans can be considered as a source of contamination of this food with oxacillin-resistant S. aureus strains. The genotypes of the investigated milk borderline-resistant isolates were shown to occur not only in cows, but also in humans. Since manual handling is reduced in raw milk production, a human origin of S. aureus seems unlikely. Because knowledge of the genotypes of animal staphylococci is limited, more research is needed to address the question of the origin of antibiotic-resistant S. aureus strains in food.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Contaminação de Alimentos/análise , Oxacilina/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Animais , Bovinos , Contagem de Colônia Microbiana , Qualidade de Produtos para o Consumidor , Relação Dose-Resposta a Droga , Genótipo , Humanos , Produtos da Carne/microbiologia , Testes de Sensibilidade Microbiana , Leite/microbiologia , Staphylococcus aureus/isolamento & purificação
17.
FEMS Microbiol Lett ; 296(1): 72-7, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19459969

RESUMO

Enterotoxigenic Staphylococcus aureus has been associated with staphylococcal food poisoning, which in a number of patients is accompanied by gastroenteritis. It has also been found to persist asymptomatically in the human intestinal tract, being considered one of the sources of pathogen transmission to manually handled food. However, very little is known about the incidence and enterotoxigenicity of intestinal S. aureus not associated with enteritis. There are practically no data on the frequency of some enterotoxin genes in intestinal S. aureus. Six thousand six hundred and twenty-one fecal swabs from 6-month- to 8-year-old children were analyzed for S. aureus. Growth of S. aureus was found in 347 samples. Two hundred and eight S. aureus isolates (4.2% of 4900 swabs) were from patients with sporadic enteritis and 139 isolates (8% of 1721 swabs) from patients who did not develop diarrhea during hospitalization. The genes encoding 16 members of the enterotoxin family (sea-see, seg-selp, and selu) and tst were present in 174 (83%) S. aureus isolates accompanying enteritis and in 101 (72%) isolates derived from patients with no enteritis symptoms. The genes of the classical enterotoxins (sea-see) and tst were present in 56% and 59% of the enteritis-associated and nonenteritic isolates, respectively.


Assuntos
Proteínas de Bactérias/genética , Enterotoxinas/genética , Fezes/microbiologia , Staphylococcus aureus/genética , Criança , Pré-Escolar , DNA Bacteriano/genética , Genótipo , Humanos , Lactente , Reação em Cadeia da Polimerase/métodos , Prevalência , Staphylococcus aureus/isolamento & purificação
18.
Med Dosw Mikrobiol ; 60(2): 101-10, 2008.
Artigo em Polonês | MEDLINE | ID: mdl-18819446

RESUMO

The aim of this study was to evaluate the frequency of isolation and antimicrobial resistance testing of non-fermentative rods isolated from clinical specimens from patients hospitalized in Korczak Pediatric Center of Lower Silesia in Wroclaw. The susceptibility of bacteria to selected antibiotics was determined. The commonest pathogens were Pseudomonas rods (81.8%) isolated from respiratory system and urine of patients hospitalized in unit intensive care. Variety of resistance patterns were detected in bacteria. ESBL were detected the most of S. maltophilia. Strains of Pseudomonas and Acinetobacter resistant to carbapenems were detected with a frequency of 5.5% and 35.9%.


Assuntos
Acinetobacter/classificação , Acinetobacter/efeitos dos fármacos , Resistência Microbiana a Medicamentos , Pseudomonas/classificação , Pseudomonas/efeitos dos fármacos , Sistema Respiratório/microbiologia , Urina/microbiologia , Acinetobacter/isolamento & purificação , Carbapenêmicos/farmacologia , Criança , Humanos , Unidades de Terapia Intensiva Pediátrica , Polônia , Pseudomonas/isolamento & purificação , Especificidade da Espécie
19.
Med Dosw Mikrobiol ; 60(4): 303-9, 2008.
Artigo em Polonês | MEDLINE | ID: mdl-19382602

RESUMO

Pseudomonas aeruginosa strains are the prominent nosocomial pathogens, especially dangerous for patients hospitalized in intensive care units. The cell surface hydrophobicity of P. aeruginosa rods and resistance to the bactericidal action of serum are considered as important factors of their virulence. The aim of the study was to evaluate the susceptibility of hydrophilic and hydrophobic P aeruginosa strains to the bactericidal effect of human serum. These strains were isolated from bronchioloalveolar lavage of children with hospital-acquired pneumonia. The BATH test was used to evaluate the hydrophobic properties. Among tested P aeruginosa strains seven had strong hydrophobic properties and eight strains were hydrophilic. The data showed that hydrophobic strains were more frequently serum-resistant than rods with hydrophilic cell surface.


Assuntos
Líquido da Lavagem Broncoalveolar/microbiologia , Infecção Hospitalar/imunologia , Infecção Hospitalar/microbiologia , Pneumonia Bacteriana/microbiologia , Infecções por Pseudomonas/imunologia , Pseudomonas aeruginosa/patogenicidade , Soro/imunologia , Criança , Infecção Hospitalar/sangue , Humanos , Interações Hidrofóbicas e Hidrofílicas , Proteínas de Membrana/imunologia , Pneumonia Bacteriana/sangue , Pneumonia Bacteriana/imunologia , Infecções por Pseudomonas/sangue , Pseudomonas aeruginosa/classificação , Especificidade da Espécie
20.
Med Dosw Mikrobiol ; 59(1): 75-84, 2007.
Artigo em Polonês | MEDLINE | ID: mdl-17601034

RESUMO

The aim of this study was to evaluate the frequency of isolation and antimicrobial resistance testing of bacterial strains isolated from clinical specimens from patients hospitalized in three Intensive Care Units in Wroclaw. The susceptibility of bacteria (107 strains) to selected antibiotics was determined. The results clearly show that non-fermentative rods were identified as the main agents causing pneumonia (58% of isolates). The second commonest pathogens were Gram-positive cocci (29%). The P. aeruginosa and E. cloacae strains were resistant to ampicillin, amoxicillin/clavulanate, cefuroxime and cefotaxime. All isolates of A. baumanii were susceptible only to imipenem. The rods of K. pneumoniae and E. coli were resistant to ampicillin, about 55% strains of both bacteria were sensitive to other antibiotics, except piperacillin/tazobactam, imipenem and ciprofloxacin. About 90% of methicillin resistant S. epidermidis strains were resistant to all antibiotics, except vancomycin (100% isolates were sensitive). ESBL were detected among E. cloace, K. pneumoniae and E. coli. We found P. aeruginosa rods producing MBL.


Assuntos
Antibacterianos/farmacologia , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana Múltipla , Bactérias Gram-Negativas/isolamento & purificação , Cocos Gram-Positivos/isolamento & purificação , Unidades de Terapia Intensiva , Infecções Respiratórias/microbiologia , Adolescente , Adulto , Idoso , Brônquios/microbiologia , Criança , Pré-Escolar , Feminino , Bactérias Gram-Negativas/efeitos dos fármacos , Cocos Gram-Positivos/efeitos dos fármacos , Humanos , Imipenem , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificação , Especificidade da Espécie , Virulência
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