RESUMO
Staphylococcus pseudintermedius, which is part of the skin microbiome of dogs, causes a variety of opportunistic infections. These infections may become more difficult to treat due to the formation of biofilm. The capacity of S. pseudintermedius to form biofilm, as well as the associated genes, has not been elucidated. This study evaluated the production and composition of S. pseudintermedius biofilm. Samples were collected from both infected dogs and asymptomatic dogs. Isolates were identified using mass spectrometry and Multiplex-PCR. Biofilm production and composition were assessed using a quantitative microtiter plate assay. The presence of ica operon genes and sps genes was investigated using conventional PCR. The investigation of Agr type and virulence genes was conducted in silico on 24 sequenced samples. All strains could produce strong biofilms, with most of the isolates presenting a polysaccharide biofilm. 63.6% of the isolates carried the complete ica operon (ADBC). All samples showed the presence of the genes spsK, spsA, and spsL, while the distribution of other genes varied. Agr type III was the most prevalent (52.2%). All sequenced samples carried the cytotoxins hlb, luk-S, luk-F, as well as the exfoliative toxins siet and se_int. No isolate displayed other exfoliative toxins. Only LB1733 presented a set of different enterotoxins (sea, seb, sec_canine, seh, sek, sel, and seq). Our findings suggest that S. pseudintermedius is a strong producer of biofilm and carries virulence genes.
Assuntos
Biofilmes , Doenças do Cão , Staphylococcus , Animais , Biofilmes/crescimento & desenvolvimento , Cães , Doenças do Cão/microbiologia , Virulência/genética , Staphylococcus/genética , Staphylococcus/isolamento & purificação , Staphylococcus/patogenicidade , Staphylococcus/classificação , Staphylococcus/fisiologia , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Fatores de Virulência/genética , Proteínas de Bactérias/genética , ÓperonRESUMO
Biofilm-producing methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci (MR-CoNS) pose clinical challenges in treating healthcare-associated infections. As alternative antimicrobial options are needed, in this study, we aimed to determine the effect of curcumin-chitosan magnetic nanoparticles (Cur-Chi-MNP) on the biofilms of staphylococcal clinical isolates. MRSA and CoNS clinical isolates were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Antimicrobial susceptibility testing was performed using the broth microdilutions. Nanoparticles were synthesized by the co-precipitation of magnetic nanoparticles (MNP) and encapsulated by the ionotropic gelation of curcumin (Cur) and chitosan (Chi). Biofilm inhibition and eradication by nanoparticles, with and without the addition of oxacillin (OXA), were assessed in Staphylococcus strains. Cur-Chi-MNP showed antimicrobial activity against planktonic cells of MRSA and MR-CoNS strains and inhibited MRSA biofilm. The addition of OXA to Cur-Chi-MNP increased the biofilm inhibition and eradication activity against all staphylococcal strains (P = 0.0007), and higher biofilm activity was observed in the early biofilm stages. Cur-Chi-MNP showed antimicrobial and biofilm inhibitory activities against S. aureus. Addition of OXA increased biofilm inhibition and eradication activity against all staphylococcal strains. A combination treatment of Cur-Chi-MNP and OXA could potentially be used to treat staphylococcal biofilm-associated infections in the early stages before the establishment of biofilm bacterial cells.
Assuntos
Antibacterianos , Biofilmes , Quitosana , Curcumina , Staphylococcus aureus Resistente à Meticilina , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas , Biofilmes/efeitos dos fármacos , Quitosana/farmacologia , Quitosana/química , Curcumina/farmacologia , Antibacterianos/farmacologia , Humanos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/tratamento farmacológico , Nanopartículas de Magnetita/química , Oxacilina/farmacologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/fisiologiaRESUMO
The Caatinga biome occurs only in Brazil and offers epidemiological conditions that should be assessed differently from other regions of Brazil and the world. Thus, the aim of this survey was to identify antimicrobial resistance, enterotoxin and biofilm production genes in Staphylococcus spp. isolated from facilities and fomites in a veterinary hospital in Caatinga biome. Samples were collected from surfaces of small animal clinical care tables (n =8), cages in the dog and cat hospitalisation sector and animals with infectious diseases (n = 21), small animal surgical centre (n =8), sterilisation sector (n =7) and stethoscopes (n = 32) by using sterile swabs. Bacterial isolation and identification, antimicrobial resistance phenotypic test and molecular detection of antimicrobial resistance, biofilm formation and enterotoxin genes were carried out. Ninety-five bacterial isolates were obtained, and 29 (30.5%) were identified as Staphylococcus spp. Overall, 13 isolates (44.8%) of six species of Staphylococcus spp. showed antimicrobial resistance profile, as well as S. haemolyticus expressed phenotypic profile of multidrug resistance. The antimicrobials with the highest resistance rates were penicillin and tetracycline. The most frequent resistance genes were blaZ and tetM, both detected in 10 (76.9%) isolates. The mecA, tetL and tetK genes had frequencies of 38.5% (5/13), 23.1% (3/13) and 15.4% (2/13), respectively. The biofilm production marker, icaD gene, was detected in one S. sciuri strain. SEE gene, which encodes enterotoxins, was detected in 15.4% (2/13) of the strains (S. pseudintermedius and S. intermedius). The occurrence of Staphylococcus spp. carrying resistance genes to diferent classes of antimicrobials, presenting MDR phenotypic pattern and carrying enterotoxins and biofim encoding genes recovered from veterinary hospital facilities and fomites in the Caatinga biome reinforce the need to implement prevention cares in veterinary practices to avoid One Health-concerning conditions.
Assuntos
Antibacterianos , Biofilmes , Enterotoxinas , Fômites , Hospitais Veterinários , Testes de Sensibilidade Microbiana , Staphylococcus , Biofilmes/crescimento & desenvolvimento , Biofilmes/efeitos dos fármacos , Staphylococcus/genética , Staphylococcus/efeitos dos fármacos , Staphylococcus/isolamento & purificação , Staphylococcus/fisiologia , Animais , Enterotoxinas/genética , Antibacterianos/farmacologia , Brasil , Fômites/microbiologia , Gatos , Cães , Farmacorresistência Bacteriana/genética , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismoRESUMO
Mastitis is a common and costly disease on dairy farms, commonly caused by Staphylococcus spp. though the various species are associated with different clinical outcomes. In the current study, we performed genomic analyses to determine the prevalence of adhesion, biofilm, and related regulatory genes in 478 staphylococcal species isolated from clinical and subclinical mastitis cases deposited in public databases. The most prevalent adhesin genes (ebpS, atl, pls, sasH and sasF) were found in both clinical and subclinical isolates. However, the ebpS gene was absent in subclinical isolates of Staphylococcus arlettae, S. succinus, S. sciuri, S. equorun, S. galinarum, and S. saprophyticus. In contrast, the coa, eap, emp, efb, and vWbp genes were present more frequently in clinical (vs. subclincal) mastitis isolates and were highly correlated with the presence of the biofim operon (icaABCD) and its transcriptional regulator, icaR. Co-phylogenetic analyses suggested that many of these adhesins, biofilm, and associated regulatory genes could have been horizontally disseminated between clinical and subclinical isolates. Our results further suggest that several adhesins, biofilm, and related regulatory genes, which have been overlooked in previous studies, may be of use for virulence profiling of mastitis-related Staphylococcus strains or as potential targets for vaccine development.
Assuntos
Adesinas Bacterianas/genética , Biofilmes/crescimento & desenvolvimento , Mastite Bovina/patologia , Infecções Estafilocócicas/patologia , Staphylococcus/genética , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bovinos , Feminino , Mastite Bovina/microbiologia , Filogenia , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Staphylococcus/fisiologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Fatores de Virulência/genéticaRESUMO
Biofilm formation is a central feature to guarantee staphylococcal persistence in hosts and is associated with several diseases that are difficult to treat. In this research paper, biofilm formation and antimicrobial susceptibility were investigated in staphylococcal strains belonging to several species. These strains were isolated from the milk of cows with subclinical mastitis and most of them were coagulase-negative, with the prevalence of Staphylococcus chromogenes. High genetic diversity was observed among the strains by pulsed field gel electrophoresis. Antimicrobial resistance was assessed by disk diffusion and more than 50% of the strains were resistant to ampicillin and penicillin G, with multi-resistance profiles (13.6%) also being observed. Most strains (65.9%) formed biofilms when cultivated in BHI supplemented with 1% glucose. Most strains (72.7%) carried the intercellular adhesion gene (icaA), while less than half (36.3%) carried the biofilm-associated protein gene (bap). Concentrations of up to 10xMIC of erythromycin and tetracycline were not sufficient to suppress cell viability in preformed biofilms. Our results revealed that a genetically diverse group of biofilm-forming Staphylococcus species can be involved in subclinical mastitis. Since high antimicrobial concentrations cannot eradicate biofilm cells in vitro, their use in dairy animals may be ineffective in controlling infections, while supporting selection of resistant microorganisms. These data reinforce the need for alternative therapies aiming at disrupting biofilms for effective disease control.
Assuntos
Biofilmes/crescimento & desenvolvimento , Farmacorresistência Bacteriana/fisiologia , Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/efeitos dos fármacos , Staphylococcus/fisiologia , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Bovinos , Coagulase/análise , Farmacorresistência Bacteriana/genética , Feminino , Variação Genética , Mastite Bovina/tratamento farmacológico , Testes de Sensibilidade Microbiana/veterinária , Infecções Estafilocócicas/microbiologia , Staphylococcus/genéticaRESUMO
Staphylococcus nepalensis is a commensal bacterium from the oral microbiota of domestic cats, with a still obscure clinical importance. In this work, we analysed the ability of feline strains of S. nepalensis to transfer antimicrobial resistance genes to Staphylococcus aureus isolated from humans through plasmids. To this end, we first analysed all publicly available genomes from cat staphylococci using computational methods to build a pan-resistome. Genes that encode resistance to erythromycin, gentamicin, mupirocin and tetracycline, common to human and cat staphylococci and previously described to be located in mobile genetic elements, were chosen for the next analyses. We studied 15 strains of S. nepalensis, which were shown to be genetically different by GTG5-PCR. As observed by disc diffusion, resistance to tetracycline was widespread (80â%), followed by resistance to erythromycin (40â%), gentamicin (27â%) and mupirocin (7â%). The strains were positive for several antimicrobial resistance genes and more than half of them harboured plasmids. The loss of plasmids and resistance genes in some strains were induced by stress with SDS. Through conjugation experiments, we observed that these plasmids can be transferred to S. aureus, thus increasing its potential to resist drug therapy. Our findings show that S. nepalensis, an underestimated inhabitant of the cat microbiota, can be a reservoir of antimicrobial resistance genes for S. aureus and, like many other staphylococci, be an overlooked and silent threat to their animal hosts and humans living with them.
Assuntos
Reservatórios de Doenças/veterinária , Farmacorresistência Bacteriana/genética , Transferência Genética Horizontal , Staphylococcus/fisiologia , Animais , Animais Domésticos , Antibacterianos/farmacologia , Gatos , Reservatórios de Doenças/microbiologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Genes Bacterianos , Variação Genética , Testes de Sensibilidade Microbiana , Plasmídeos/efeitos dos fármacos , Plasmídeos/genética , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/efeitos dos fármacos , Staphylococcus/genética , Staphylococcus/isolamento & purificação , Staphylococcus aureus/genéticaRESUMO
Polymicrobial infections are one of the most common reasons for inflammation of surrounding tissues and failure of implanted biomaterials. Because microorganism adhesion is the first step for biofilm formation, physical-chemical modifications of biomaterials have been proposed to reduce the initial microbial attachment. Thus, the use of superhydrophobic coatings has emerged because of their anti-biofilm properties. However, these coatings on the titanium (Ti) surface have been developed mainly by dual-step surface modification techniques and have not been tested using polymicrobial biofilms. Therefore, we developed a one-step superhydrophobic coating on the Ti surface by using a low-pressure plasma technology to create a biocompatible coating that reduces polymicrobial biofilm adhesion and formation. The superhydrophobic coating on Ti was created by the glow discharge plasma using Ar, O2, and hexamethyldisiloxane gases, and after full physical, chemical, and biological characterizations, we evaluated its properties regarding oral biofilm inhibition. The newly developed coating presented an increased surface roughness and, consequently, superhydrophobicity (contact angle over 150°) and enhanced corrosion resistance (p < 0.05) of the Ti surface. Furthermore, proteomic analysis showed a unique pattern of protein adsorption on the superhydrophobic coating without drastically changing the biologic processes mediated by proteins. Additionally, superhydrophobic treatment did not present a cytotoxic effect on fibroblasts or reduction of proliferation; however, it significantly reduced (≈8-fold change) polymicrobial adhesion (bacterial and fungal) and biofilm formation in vitro. Interestingly, superhydrophobic coating shifted the microbiological profile of biofilms formed in situ in the oral cavity, reducing by up to ≈7 fold pathogens associated with the peri-implant disease. Thus, this new superhydrophobic coating developed by a one-step glow discharge plasma technique is a promising biocompatible strategy to drastically reduce microbial adhesion and biofilm formation on Ti-based biomedical implants.
Assuntos
Materiais Revestidos Biocompatíveis/química , Implantes Dentários/microbiologia , Titânio/química , Animais , Aderência Bacteriana , Biofilmes , Candida albicans/fisiologia , Sobrevivência Celular , Corrosão , Fibroblastos/citologia , Interações Hidrofóbicas e Hidrofílicas , Camundongos , Staphylococcus/fisiologia , Propriedades de SuperfícieRESUMO
Common alloys used for the manufacture of aircrafts are subject to different forms of environmental deterioration. A major one is corrosion, and there is a strong body of evidence suggesting that environmental microorganisms initiate and accelerate it. The development of an appropriate strategy to reduce this process depends on the knowledge concerning the factors involved in corrosion. In this work, a biofilm forming bacterial consortium was extracted in situ from the corrosion products formed in an aircraft exposed to Antarctic media. Two thermophilic bacteria, an Anoxybacillus and a Staphylococcus strain, were successfully isolated from this consortium. Two extracellular enzymes previously speculated to participate in corrosion, catalase and peroxidase, were detected in the extracellular fraction of the consortium. Additionally, we assessed the individual contribution of those thermophilic microorganisms on the corrosion process of 7075-T6 aluminum alloy, which is widely used in aeronautical industry, through electrochemical methods and surface analysis techniques.
Assuntos
Ligas/química , Alumínio/química , Anoxybacillus/fisiologia , Biofilmes , Anoxybacillus/enzimologia , Anoxybacillus/isolamento & purificação , Regiões Antárticas , Corrosão , Oxirredução , Staphylococcus/enzimologia , Staphylococcus/isolamento & purificação , Staphylococcus/fisiologia , Propriedades de SuperfícieRESUMO
This review aimed to describe the biofilm formation ability of coagulase-negative Staphylococcus, addressing its impact to the food industry. Coagulase-negative Staphylococcus have the ability to produce enterotoxins in food, making it an important line of study, as it constitutes a risk to public health. The biofilm formation by these microorganisms requires physicochemical processes, such as hydrophobic forces, which are essential for the first phase of fixing the biofilm on the surface. In industrial facilities, stainless steel equipment is the most associated with the formation of biofilms, due to the presence grooves and cracks. Many species of coagulase-negative Staphylococcus produce biofilm, but the most studied is S. epidermidis, as it is the most frequently isolated from food. Coagulase-negative Staphylococcus form biofilm on different surfaces in the food industry, and can become a source of permanent contamination, that can be present in the final product, intended for human consumption. Among other alternatives to combat the formation of biofilm in industrial food facilities, there is the implementation of Good Manufacturing Practices, which is effective in preventing bacterial adhesion, and therefore, the formation of biofilm. However, further studies are needed in order to quantify the occurrence of coagulase-negative Staphylococcus biofilms in the food industry.(AU)
Esta revisão bibliográfica teve como objetivo descrever a capacidade de formação de biofilme por Staphylococcus coagulase-negativo, relacionando seu impacto na indústria alimentícia. Os Staphylococcus coagulase-negativos possuem a capacidade de produzir enterotoxina nos alimentos, tornando-se uma importante linha de estudo, pois constitui um risco para a saúde pública. A formação do biofilme por esses micro-organismos requer processos físico-químicos, como forças hidrofóbicas, essenciais para a primeira fase de fixação do biofilme na superfície. Nas indústrias, equipamentos de aço inoxidável são os mais associados à formação de biofilmes, em decorrência de possuírem ranhuras e fendas. Muitas espécies de Staphylococcus coagulase-negativo produzem biofilme, porém, o mais estudado é o S. epidermidis, por ser o mais frequentemente isolado de alimentos. Os Staphylococcus coagulase-negativos formam biofilme em diferentes superfícies de indústrias alimentícias, podendo se tornar uma fonte de contaminação permanente, contaminando o produto final destinado ao consumo humano. Dentre outras alternativas para combater a formação do biofilme nas plantas alimentícias, a implantação das Boas Práticas de Fabricação é eficaz para prevenir a adesão bacteriana, evitando a formação do biofilme. No entanto, são necessários estudos para quantificar a ocorrência de biofilmes de Staphylococcus coagulase-negativos em indústrias alimentícias.(AU)
Assuntos
Staphylococcus/fisiologia , Coagulase , Biofilmes/crescimento & desenvolvimento , Contaminação de Alimentos , Indústria AlimentíciaRESUMO
Introduction: Infections associated with health care caused by S. aureus and coagulase-negative Staphylococci multi-resistant to antibiotics cause a high epidemiological impact due to their high morbidity and mortality. Biofilm formation, which has been associated with antimicrobial resistance, can also occur. Objectives: To determine methicillin resistance and to quantify the biofilm production to establish if there is a relationship in clinical isolates of S. aureus and coagulase-negative Staphylococci. Material and methods: A total of 11 strains of S. aureus and 12 of coagulase-negative Staphylococci were studied. Methicillin resistance was determined with cefoxitin discs and the Clinical Laboratory Standards Institute (CSLI), 2018 reference values. Biofilm production was quantified by the crystal violet method. The mecA and icaADBC genes were identified by PCR. A bivariate analysis was performed with chi-square (c2) and Cramér's V statistical tests, using SPSS™, version 20.0 software. Results: Nine S. aureus strains were methicillin-resistant and two were sensitive. Eight coagulase-negative Staphylococci strains were resistant and four were sensitive. The mecA genotype was found in eight of the nine S. aureus resistant strains and six of eight resistant coagulase-negative Staphylococci. All strains formed biofilms. Ten strains of S. aureus and 11 of coagulase-negative Staphylococci presented the icaADCB genotype. No association was found between methicillin-resistance and biofilm formation. Conclusions: Cefoxitin is enough to define the resistance phenotype and is associated with the mecA genotype. All strains formed biofilms and were related to the presence of the icaADCB operon. Biofilm formation and methicillin resistance were independent features in both groups of strains.
Introducción. Las infecciones por Staphylococcus aureus y Staphylococcus coagulasa negativa multirresistentes a los antibióticos y asociadas con la atención en salud tienen un gran impacto epidemiológico por su alta morbimortalidad; además, se han relacionado con la formación de biopelículas, lo cual también se asocia con la resistencia a los antimicrobianos. Objetivo. Determinar la resistencia a la meticilina y cuantificar la producción de biopelículas para establecer su posible relación con los aislamientos clínicos de S. aureus y Staphylococcus coagulasa negativa. Materiales y métodos. Se estudiaron 11 cepas de S. aureus y 12 de Staphylococcus coagulasa negativa. La resistencia a la meticilina se determinó con discos de cefoxitina tomando como valores de referencia los estándares del Clinical Laboratory Standards Institute (CLSI) de 2018. La producción de biopelícula se cuantificó con cristal violeta. Los genes mecA e icaADBC se identificaron mediante reacción en cadena de la polimerasa (PCR), y se hizo un análisis bivariado con la prueba de ji al cuadrado y el coeficiente V de Cramér, utilizando el programa SPSS™, versión 20.0. Resultados. Nueve cepas de S. aureus fueron resistentes a la meticilina (SARM) y dos fueron sensibles. Ocho cepas de Staphylococcus coagulasa negativa fueron resistentes y cuatro fueron sensibles. El genotipo mecA se encontró en ocho de las nueve cepas de S. aureus y en seis de las ocho de Staphylococcus coagulasa negativa resistentes a meticilina. Todas las cepas formaron biopelícula. Diez cepas de S. aureus y 11 de Staphylococcus coagulasa negativa presentaron el genotipo icaADCB. No se encontró asociación entre la resistencia a meticilina y la formación de biopelícula. Conclusiones. La cefoxitina es suficiente para determinar el fenotipo resistente a meticilina y se asoció con el genotipo mecA. Las cepas resistentes a la meticilina y poseedoras del gen mecA pueden presentar un mecanismo de resistencia alterno. Los dos grupos de cepas formadoras de biopelícula se relacionaron con la presencia del operón icaADCB. La formación de biopelícula y la resistencia a la meticilina se expresaron como características independientes en los dos grupos de cepas.
Assuntos
Biofilmes/crescimento & desenvolvimento , Resistência a Meticilina , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/fisiologia , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Cefoxitina/farmacologia , Coagulase , DNA Bacteriano/isolamento & purificação , Genes Bacterianos , Humanos , Resistência a Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , México , Testes de Sensibilidade Microbiana/métodos , Oxacilina/farmacologia , Proteínas de Ligação às Penicilinas/genética , Infecções Estafilocócicas/microbiologia , Staphylococcus/enzimologia , Staphylococcus/genética , Staphylococcus aureus/genéticaRESUMO
Resumen Introducción. Las infecciones por Staphylococcus aureus y Staphylococcus coagulasa negativa multirresistentes a los antibióticos y asociadas con la atención en salud tienen un gran impacto epidemiológico por su alta morbimortalidad; además, se han relacionado con la formación de biopelículas, lo cual también se asocia con la resistencia a los antimicrobianos. Objetivo. Determinar la resistencia a la meticilina y cuantificar la producción de biopelículas para establecer su posible relación con los aislamientos clínicos de S. aureus y Staphylococcus coagulasa negativa. Materiales y métodos. Se estudiaron 11 cepas de S. aureus y 12 de Staphylococcus coagulasa negativa. La resistencia a la meticilina se determinó con discos de cefoxitina tomando como valores de referencia los estándares del Clinical Laboratory Standards Institute (CLSI) de 2018. La producción de biopelícula se cuantificó con cristal violeta. Los genes mecA e icaADBC se identificaron mediante reacción en cadena de la polimerasa (PCR), y se hizo un análisis bivariado con la prueba de ji al cuadrado y el coeficiente V de Cramér, utilizando el programa SPSS™, versión 20.0. Resultados. Nueve cepas de S. aureus fueron resistentes a la meticilina (SARM) y dos fueron sensibles. Ocho cepas de Staphylococcus coagulasa negativa fueron resistentes y cuatro fueron sensibles. El genotipo mecA se encontró en ocho de las nueve cepas de S. aureus y en seis de las ocho de Staphylococcus coagulasa negativa resistentes a meticilina. Todas las cepas formaron biopelícula. Diez cepas de S. aureus y 11 de Staphylococcus coagulasa negativa presentaron el genotipo icaADCB. No se encontró asociación entre la resistencia a meticilina y la formación de biopelícula. Conclusiones. La cefoxitina es suficiente para determinar el fenotipo resistente a meticilina y se asoció con el genotipo mecA. Las cepas resistentes a la meticilina y poseedoras del gen mecA pueden presentar un mecanismo de resistencia alterno. Los dos grupos de cepas formadoras de biopelícula se relacionaron con la presencia del operón icaADCB. La formación de biopelícula y la resistencia a la meticilina se expresaron como características independientes en los dos grupos de cepas.
Abstract Introduction: Infections associated with health care caused by S. aureus and coagulase- negative Staphylococci multi-resistant to antibiotics cause a high epidemiological impact due to their high morbidity and mortality. Biofilm formation, which has been associated with antimicrobial resistance, can also occur. Objectives: To determine methicillin resistance and to quantify the biofilm production to establish if there is a relationship in clinical isolates of S. aureus and coagulase-negative Staphylococci. Material and methods: A total of 11 strains of S. aureus and 12 of coagulase-negative Staphylococci were studied. Methicillin resistance was determined with cefoxitin discs and the Clinical Laboratory Standards Institute (CSLI), 2018 reference values. Biofilm production was quantified by the crystal violet method. The mecA and icaADBC genes were identified by PCR. A bivariate analysis was performed with chi-square (c2) and Cramér's V statistical tests, using SPSS™, version 20.0 software. Results: Nine S. aureus strains were methicillin-resistant and two were sensitive. Eight coagulase-negative Staphylococci strains were resistant and four were sensitive. The mecA genotype was found in eight of the nine S. aureus resistant strains and six of eight resistant coagulase-negative Staphylococci. All strains formed biofilms. Ten strains of S. aureus and 11 of coagulase-negative Staphylococci presented the icaADCB genotype. No association was found between methicillin-resistance and biofilm formation. Conclusions: Cefoxitin is enough to define the resistance phenotype and is associated with the mecA genotype. All strains formed biofilms and were related to the presence of the icaADCB operon. Biofilm formation and methicillin resistance were independent features in both groups of strains.
Assuntos
Humanos , Staphylococcus/efeitos dos fármacos , Staphylococcus/fisiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Resistência a Meticilina , Biofilmes/crescimento & desenvolvimento , Oxacilina/farmacologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/enzimologia , Staphylococcus/genética , Staphylococcus aureus/genética , Proteínas de Bactérias/genética , DNA Bacteriano/isolamento & purificação , Testes de Sensibilidade Microbiana/métodos , Cefoxitina/farmacologia , Resistência a Meticilina/genética , Coagulase , Proteínas de Ligação às Penicilinas/genética , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , Genes Bacterianos , México , Antibacterianos/farmacologiaRESUMO
Objective: To determine the prevalence and virulence factors of coagulase-negative Staphylococci (CNS) in prosthetic joint infections (PJI). Method: CNS were isolated of 66 hip and knee PJI from Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, México City. Antimicrobial susceptibility and biofilm formation in CNS were determined; icaADBC, aap, bap and embp genes were determined by PCR. Results: Staphylococcus and Staphylococcus hominis were the most prevalent with 82 y 80% respectively. Staphylococcus lugdunensis, Staphylococcus haemolyticus, Staphylococcus capitis, Staphylococcus caprae, Staphylococcus sciuri and Staphylococcus lentus were less frequent. The majority of isolates were resistant to ß-lactam antibiotics, fluoroquinolone, and erythromycin. 41% of CNS were biofilm former and 59% were non-biofilm former (p = 0.0551). Biofilm former Staphylococcus epidermidis showed a high presence of icaADBC, aap and embp operons compared to the non-biofilm former isolates (p < 0.05). In contrast, non-S. epidermidis CNS had only the aap gen. Conclusion: S. haemolyticus, S. sciuri and S. lentus are new isolates of PJI not previously reported with virulence factors similar to CNS isolates.
Objetivo: Estudiar la prevalencia y los factores de virulencia de Staphylococcus coagulasa negativos (SCN) de infecciones de prótesis articular (IPA). Método: Los SCN se aislaron de 66 pacientes con IPA de cadera y rodilla procedentes del Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, de Ciudad de México. Se determinaron la sensibilidad antimicrobiana y la producción de biopelículas de los SCN. Los genes icaADBC, aap, bap y embp fueron detectados por reacción en cadena de la polimerasa en SCN. Resultados: La IPA de cadera fue el 80%. Se aislaron en alta proporción S. epidermidis (82%) y S. hominis (80%), y en baja frecuencia S. lugdunensis, S. haemolyticus, S. capitis, S. caprae, S. sciuri y S. lentus. La mayoría de los aislamientos fueron resistentes a los betalactámicos, las fluoroquinolonas y la eritromicina. La producción de biopelículas se determinó en el 41% de los SCN y el 59% fueron no productores de biopelículas (p = 0.0551). S. epidermidis productores de biopelículas tuvieron mayor presencia del operón icaADBC, aap y embp que los aislamientos no productores de biopelícula (p < 0.05). Los SCN no S. epidermidis presentaron únicamente el gen aap. Conclusiones: S. haemolyticus, S. sciuri y S. lentus son aislamientos nuevos de IPA no reportados que poseen factores de virulencia, igual que las otras especies de SCN aisladas.
Assuntos
Biofilmes/crescimento & desenvolvimento , Prótese do Joelho/microbiologia , Infecções Relacionadas à Prótese/microbiologia , Staphylococcus/isolamento & purificação , Fatores de Virulência/isolamento & purificação , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias/genética , Coagulase , Farmacorresistência Bacteriana Múltipla/genética , Feminino , Genes Bacterianos , Prótese de Quadril/microbiologia , Prótese de Quadril/estatística & dados numéricos , Hospitais Especializados , Humanos , Masculino , México , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Ortopedia , Estudos Retrospectivos , Staphylococcus/classificação , Staphylococcus/enzimologia , Staphylococcus/fisiologia , Adulto JovemRESUMO
Biofilme é uma comunidade organizada de microrganismos que se forma em superfícies mal higienizadas, constituindo um mecanismo de defesa microbiana para permanência no ambiente. O objetivo do presente estudo foi investigar a capacidade de formação de biofilme de espécies de Staphylococcus coagulase negativa (SCN) isoladas de queijo Mussarela fatiado e de fatiadores de frios de estabelecimentos do município de Garanhuns-PE. De 103 isolados de SCN 56 (54,4%) foram negativos para produção de biofilme e 47 (45,6%) positivos, com a maior frequência de detecção nas espécies S. saprophyticus e S. cohnii subsp. urealyticum. Os resultados apontam para o potencial risco de contaminação cruzada de outros alimentos, uma vez que cepas bacterianas produtoras de biofilmes podem colonizar e persistir em superfícies de diversos equipamentos.(AU)
Assuntos
Biofilmes , Staphylococcus/fisiologia , Queijo/microbiologia , Contaminação de Alimentos , Contaminação de Equipamentos , Staphylococcus/isolamento & purificação , Microbiologia de Alimentos , Equipamentos para AlimentosRESUMO
Biofilme é uma comunidade organizada de microrganismos que se forma em superfícies mal higienizadas, constituindo um mecanismo de defesa microbiana para permanência no ambiente. O objetivo do presente estudo foi investigar a capacidade de formação de biofilme de espécies de Staphylococcus coagulase negativa (SCN) isoladas de queijo Mussarela fatiado e de fatiadores de frios de estabelecimentos do município de Garanhuns-PE. De 103 isolados de SCN 56 (54,4%) foram negativos para produção de biofilme e 47 (45,6%) positivos, com a maior frequência de detecção nas espécies S. saprophyticus e S. cohnii subsp. urealyticum. Os resultados apontam para o potencial risco de contaminação cruzada de outros alimentos, uma vez que cepas bacterianas produtoras de biofilmes podem colonizar e persistir em superfícies de diversos equipamentos.
Assuntos
Biofilmes , Contaminação de Alimentos , Contaminação de Equipamentos , Queijo/microbiologia , Staphylococcus/fisiologia , Staphylococcus/isolamento & purificação , Equipamentos para Alimentos , Microbiologia de AlimentosRESUMO
Bovine mastitis affects the health of dairy cows and the profitability of herds worldwide. Coagulase-negative staphylococci (CNS) are the most frequently isolated pathogens in bovine intramammary infection. Based on the wide range of antimicrobial, mucoadhesive and immunostimulant properties demonstrated by chitosan, we have evaluated therapy efficiency of chitosan incorporation to cloxacillin antibiotic as well as its effect against different bacterial lifestyles of seven CNS isolates from chronic intramammary infections. The therapeutic effects of combinations were evaluated on planktonic cultures, bacterial biofilms and intracellular growth in mammary epithelial cells. We found that biofilms and intracellular growth forms offered a strong protection against antibiotic therapy. On the other hand, we found that chitosan addition to cloxacillin efficiently reduced the antibiotic concentration necessary for bacterial killing in different lifestyle. Remarkably, the combined treatment was not only able to inhibit bacterial biofilm establishment and increase preformed biofilm eradication, but it also reduced intracellular bacterial viability while it increased IL-6 secretion by infected epithelial cells. These findings provide a new approach to prophylactic drying therapy that could help to improve conventional antimicrobial treatment against different forms of bacterial growth in an efficient, safer and greener manner reducing multiresistant bacteria generation and spread.
Assuntos
Antibacterianos/uso terapêutico , Quitosana/uso terapêutico , Cloxacilina/uso terapêutico , Mastite Bovina/tratamento farmacológico , Infecções Estafilocócicas/veterinária , Staphylococcus/efeitos dos fármacos , Animais , Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Bovinos , Quitosana/administração & dosagem , Quitosana/farmacologia , Cloxacilina/administração & dosagem , Cloxacilina/farmacologia , Feminino , Mastite Bovina/microbiologia , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia , Staphylococcus/isolamento & purificação , Staphylococcus/fisiologiaRESUMO
Bovine mastitis, considered the most important cause of economic losses in the dairy industry, is a major concern in veterinary medicine. Staphylococcus aureus and coagulase-negative staphylococci (CNS) are the main pathogens associated with intramammary infections, and bacterial biofilms are suspected to be responsible for the persistence of this disease. CNS from the udder are not necessarily associated with intramammary infections. In fact, some commensal CNS have been shown to have biological activities. This issue led us to screen exoproducts from commensal Staphylococcus chromogenes for anti-biofilm activity against different mastitis pathogens. The cell-free supernatant from S. chromogenes LN1 (LN1-CFS) was confirmed to display a non-biocidal inhibition of pathogenic biofilms. The supernatant was subjected to various treatments to estimate the nature of the biofilm-inhibiting compounds. The results showed that the bioactive compound >5KDa in mass is sensitive to thermal treatment and proteinase K digestion, suggesting its protein properties. LN1-CFS was able to significantly inhibit S. aureus and CNS biofilm formation in a dose-independent manner and without affecting the viability of bovine cells. These findings reveal a new activity of the udder microflora of healthy animals. Studies are underway to purify and identify the anti-biofilm biocompound and to evaluate its biological activity in vivo.
Assuntos
Biofilmes/crescimento & desenvolvimento , Mastite Bovina/microbiologia , Staphylococcus/classificação , Staphylococcus/fisiologia , Animais , Bovinos , Sobrevivência Celular , Feminino , Glândulas Mamárias Animais/citologia , Glândulas Mamárias Animais/microbiologia , Leite/microbiologia , FilogeniaRESUMO
Mastitis affects the health and welfare of dairy cows worldwide. Coagulase-negative staphylococci (CNS) are known to form biofilms and are increasingly recognized as a cause of persistent bovine intramammary infections. A total of 90 CNS isolated from cows with clinical and subclinical mastitis in Argentina from 2008 to 2014 were identified by PCR-RFLP using the gap gene. Standard microtiter plate assays were used to assess CNS biofilm formation, and Staphylococcus haemolyticus species formed the strongest biofilms. The presence of biofilm-associated genes icaA, bap and aap was detected in a few isolates, while embP, fbe, atlE and eno were present in the majority of isolates. Genes encoding resistance to ß-lactams were detected among the isolates; blaZ, mecA and mecC were detected in 21, 4 and 1 isolate, respectively. Resistance to macrolides and lincosamides (n = 6) was attributable to ermB, ermC, mphC or mrsA or a combination of those genes. In this study, we identified CNS species involved in mastitis and provide information about pathogenicity and antimicrobial resistance, which is essential to design efficient strategies to control mastitis caused by CNS.
Assuntos
Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Farmacorresistência Bacteriana Múltipla/genética , Mastite Bovina/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/fisiologia , Animais , Argentina , Bovinos , Coagulase/genética , Coagulase/metabolismo , Feminino , Genes Bacterianos , Testes de Sensibilidade Microbiana , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase em Tempo Real , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/genética , Staphylococcus/isolamento & purificaçãoRESUMO
INTRODUCTION: Biofilm-associated antimicrobial resistance is of increasing importance to the maintenance and spread of foodborne pathogens in the food industry. This study aimed to investigate the ability to form biofilm and the antimicrobial resistance of staphylococci contaminating small-scale goat milk dairy plants. METHODOLOGY: Sixty isolates were tested for antimicrobial resistance against 20 drugs by the microdilution method. Biofilm-forming traits were assessed by the microtiter plate method (MtP), Congo red agar method (CRA), and icaD gene detection by polymerase chain reaction (PCR). RESULTS: High antimicrobial resistance to ampicillin (60/60; 100%), penicillin G (21/60; 35%), and erythromycin (15/60; 25%) was observed, but all isolates were susceptible to amoxicillin/K-clavulanate, ceftriaxone, ciprofloxacin, gentamicin, levofloxacin, linezolid, and moxifloxacin. No resistance to oxacillin or vancomycin was seen among Staphylococcus aureus. Twenty-seven isolates (27/60; 45%) were considered to form biofilm according to MtP, and similar biofilm-producing frequencies were observed in coagulase-negative staphylococci (CoNS) (20/44; 45.4%) and S. aureus (7/16; 43.7%). The icaD gene was observed only in S. aureus isolates. There was no association between biofilm production and antimicrobial resistance. A higher frequency of biofilm-producing staphylococci was found in isolates from bulk tank milk and hand swabs. On the other hand, isolates from pasteurized milk showed lower frequency of biofilm formation. CONCLUSIONS: Staphylococci contaminating goat dairy plants are potential biofilm producers. The results suggest no association between the ability to form biofilm and antimicrobial resistance.
Assuntos
Biofilmes/crescimento & desenvolvimento , Farmacorresistência Bacteriana , Microbiologia Ambiental , Mãos/microbiologia , Leite/microbiologia , Staphylococcus/isolamento & purificação , Staphylococcus/fisiologia , Animais , Antibacterianos/farmacologia , Técnicas Bacteriológicas , Indústria de Laticínios , Cabras , Humanos , Reação em Cadeia da Polimerase , Staphylococcus/efeitos dos fármacosRESUMO
Coagulase-negative staphylococci (CNS) represent one of the most prevalent microorganisms in nosocomial infections worldwide, nevertheless little is known about their pathogenicity features. Thus, our aim was to characterize virulence aspects of CNS isolated from patients with bloodstream infections assisted in hospitals of Belo Horizonte, MG, Brazil. Strains were identified using bioMérieuxVitek® and for biofilm production evaluation, Congo Red Agar (CRA) and polystyrene plates were used. PCR was applied to detect icaA, icaB, icaC, atlE, sea, sec, sed, tsst-1 and agr. For statistical analyses were used hierarchical cluster, chi-square test and correspondence. 59 strains were analyzed, being S. haemolyticus the most prevalent. On CRA, 96.5% were biofilm producer, whereas on polystyrene plate, 100% showed adhesion at different times evaluated. Regarding genotypic analyses, 15.2%, 38.9%, 8.4%, 49.1%, 76.2%, 23.7%, 1.6%, 30.5% and 38.9% were positive for icaA, icaB, icaC, atlE, sea, sec, sed, tsst-1 and agr, respectively. Six clusters were formed and frequency distributions of agr, atlE, icaA, icaB, sea, sec, tsst-1 differed (P < 0.001). In conclusion, all strains were biofilm producer, with high prevalence of atlE, and had potential of toxin production, with high prevalence of sea. According to the group-analyses, icaB showed relationship with the strong adherence in samples.
Assuntos
Toxinas Bacterianas/análise , Biofilmes/crescimento & desenvolvimento , Sepse/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/fisiologia , Aderência Bacteriana , Toxinas Bacterianas/genética , Técnicas de Tipagem Bacteriana , Brasil , Análise por Conglomerados , Infecção Hospitalar/microbiologia , Genótipo , Hospitais , Humanos , Reação em Cadeia da Polimerase , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Staphylococcus/metabolismo , Fatores de Virulência/análise , Fatores de Virulência/genéticaRESUMO
Platelet Concentrates (PCs) are the blood components with the highest rate of bacterial contamination, and coagulase-negative staphylococci (CoNS) are the most frequently isolated contaminants. This study investigated the biofilm formation of 16 contaminated units out of 691 PCs tested by phenotypic and genotypic methods. Adhesion in Borosilicate Tube (ABT) and Congo Red Agar (CRA) tests were used to assess the presence of biofilm. The presence of icaADC genes was assessed by means of the Polymerase Chain Reaction (PCR) technique. With Vitek(r)2, Staphylococcus haemolyticus was considered the most prevalent CoNS (31.25%). The CRA characterized 43.8% as probable biofilm producers, and for the ABT test, 37.5%. The icaADC genes were identified in seven samples by the PCR. The ABT technique showed 85.7% sensitivity and 100% specificity when compared to the reference method (PCR), and presented strong agreement (k = 0.8). This study shows that species identified as PCs contaminants are considered inhabitants of the normal skin flora and they might become important pathogens. The results also lead to the recommendation of ABT use in laboratory routine for detecting biofilm in CoNS contaminants of PCs.