RESUMO
Staphylococcus aureus are extremely important microorganisms, either from an epidemiological point of view or as a pathogen, responsible for causing a series of infectious processes, whether simple, restricted to the skin, or invasive infections such as bacteremia. The emergence of Oxacillin Sensitive-Methicillin Resistant S.aureus (OS-MRSA) isolates has imposed difficulties in the treatment of patients with staphylococcal infection, as such isolates can be mistakenly classified as sensitive and lead to failure of the therapy used. Thus, the objective of this study is to evaluate the prevalence, and genotypic and phenotypic characteristics, of OS-MRSA isolates, from bloodstream infections, collected from patients admitted to a hospital in southern Brazil, as well as to evaluate the treatment used. For this, 801 unique isolates of S. aureus, collected from blood cultures, between January 2011 and December 2020 were evaluated. Of these, 96 isolates were classified as sensitive to oxacillin. The isolates were identified and had their sensitivity profile performed by manual and automated methods. The minimum inhibitory concentration for vancomycin, daptomycin, oxacillin, linezolid and teicoplanine was performed by e-test. The mecA, vanA genes, typing of the SCCmec elements, as well as the search for the icaA, tst-1 and pvl virulence genes were performed by PCR. Biofilm formation was performed using the crystal violet technique. The Sequence Type (ST), as well as the Clonal Complex (CC) of the isolates was evaluated by the RTq -PCR. The clinical characteristics of the patients were evaluated through an active search in medical records. After investigating the mecA gene, 27.1% (26/96) of the isolates were considered OS-MRSA, with SCCmec type I being the most prevalent, 46.1% (12/26). Among the evaluated isolates, 41% (9/22) were included in CC5 and ST9. As for virulence, all isolates were positive for the icaA gene and characterized as strong biofilm formers. The pvl gene was found in 92.3% (24/26) of the isolates and the toxic shock syndrome toxin was present in 61.5% of the isolates (16/26). All isolates were negative for the presence of the van A gene. As for the clinical outcome, 73% (19/26) of the patients were discharged from the hospital and 27% (7/26) died. It was possible to observe a high frequency of OS-MRSA isolates causing bloodstream infections. Furthermore, such isolates contain several virulence genes, which may contribute to a worse clinical outcome.
Assuntos
Antibacterianos , Bacteriemia , Proteínas de Bactérias , Genótipo , Staphylococcus aureus Resistente à Meticilina , Testes de Sensibilidade Microbiana , Oxacilina , Proteínas de Ligação às Penicilinas , Infecções Estafilocócicas , Centros de Atenção Terciária , Humanos , Oxacilina/farmacologia , Brasil , Antibacterianos/farmacologia , Infecções Estafilocócicas/microbiologia , Centros de Atenção Terciária/estatística & dados numéricos , Bacteriemia/microbiologia , Proteínas de Bactérias/genética , Proteínas de Ligação às Penicilinas/genética , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/classificação , FenótipoRESUMO
Staphylococcus aureus is a common clinical pathogen that causes various human infections. The aim of this study was to investigate the antibiotic susceptibility pattern, molecular epidemiological characteristics, and biofilm formation ability of S. aureus isolates from clinical specimens in Xiangyang and to analyze the correlation among them. A total of 111 non-duplicate S. aureus isolates were collected from the Affiliated Hospital of Hubei University of Arts and Science. All isolates were tested for antibacterial susceptibility. Methicillin-resistant S. aureus (MRSA) was identified by the mecA gene PCR amplification. All isolates were analyzed to determine their biofilm-forming ability using the microplate method. The biofilm-related gene was determined using PCR. SCCmec, MLST, and spa types of MRSA strains were performed to ascertain the molecular characteristics. Among the 111 S. aureus isolates, 45 (40.5%) and 66 (59.5%) were MRSA and MSSA, respectively. The resistance of MRSA strains to the tested antibiotics was significantly stronger than that of MSSA strains. All isolates were able to produce biofilm with levels ranging from strong (28.9%, 18.2%), moderate (62.2%, 62.1%), to weak (8.9%, 19.7%). Strong biofilm formation was observed in MRSA strains than in MSSA strains, based on percentages. There were dynamic changes in molecular epidemic characteristics of MRSA isolates in Xiangyang. SCCmecIVa-ST22-t309, SCCmecIVa-ST59-t437, and SCCmecIVa-ST5-t2460 were currently the main epidemic clones in this region. SCCmecIVa-ST5-t2460 and SCCmecIVa/III-ST22-t309 have stronger antibiotic resistance than SCCmecIVa-ST59-t437 strains, with resistance to 6 ~ 8 detected non-ß-lactam antibiotics. The molecular epidemic and resistance attributes of S. aureus should be timely monitored, and effective measures should be adopted to control the clinical infection and spread of the bacteria.
Assuntos
Antibacterianos , Biofilmes , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Infecções Estafilocócicas , Staphylococcus aureus , Centros de Atenção Terciária , Biofilmes/crescimento & desenvolvimento , Biofilmes/efeitos dos fármacos , Humanos , China/epidemiologia , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/epidemiologia , Antibacterianos/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/fisiologia , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/fisiologia , Feminino , Masculino , Proteínas de Bactérias/genética , Adulto , Farmacorresistência Bacteriana , Pessoa de Meia-Idade , Adulto Jovem , Adolescente , Tipagem de Sequências Multilocus , Criança , Idoso , Proteínas de Ligação às PenicilinasRESUMO
Bacterial cell wall formation is essential for cellular survival and morphogenesis. The peptidoglycan (PG), a heteropolymer that surrounds the bacterial membrane, is a key component of the cell wall, and its multistep biosynthetic process is an attractive antibacterial development target. Penicillin-binding proteins (PBPs) are responsible for cross-linking PG stem peptides, and their central role in bacterial cell wall synthesis has made them the target of successful antibiotics, including ß-lactams, that have been used worldwide for decades. Following the discovery of penicillin, several other compounds with antibiotic activity have been discovered and, since then, have saved millions of lives. However, since pathogens inevitably become resistant to antibiotics, the search for new active compounds is continuous. The present review highlights the ongoing development of inhibitors acting mainly in the transpeptidase domain of PBPs with potential therapeutic applications for the development of new antibiotic agents. Both the critical aspects of the strategy, design, and structure-activity relationships (SAR) are discussed, covering the main published articles over the last 10 years. Some of the molecules described display activities against main bacterial pathogens and could open avenues toward the development of new, efficient antibacterial drugs.
Assuntos
Antibacterianos , beta-Lactamas , Proteínas de Ligação às Penicilinas/química , Proteínas de Ligação às Penicilinas/metabolismo , Antibacterianos/farmacologia , beta-Lactamas/química , beta-Lactamas/farmacologia , Penicilinas/química , Penicilinas/metabolismo , Penicilinas/farmacologia , Bactérias/metabolismo , Proteínas de Bactérias/químicaRESUMO
The presence of Staphylococcus spp. resistant to methicillin in the nasal cavity of swine has been previously reported. Considering the possible occurrence of bacterial resistance and presence of resistance genes in intensive swine breeding and the known transmissibility and dispersion potential of such genes, this study aimed to investigate the prevalence of resistance to different antibiotics and the presence of the mecA resistance gene in Staphylococcus spp. from piglets recently housed in a nursery. For this, 60 nasal swabs were collected from piglets at the time of their housing in the nursery, and then Staphylococcus spp. were isolated and identified in coagulase-positive (CoPS) and coagulase-negative (CoNS) isolates. These isolates were subjected to the disk-diffusion test to evaluate the bacterial resistance profile and then subjected to molecular identification of Staphylococcus aureus and analyses of the mecA gene through polymerase chain reaction. Of the 60 samples collected, 60 Staphylococcus spp. were isolated, of which 38 (63.33%) were classified as CoNS and 22 (36.67%) as CoPS. Of these, ten (45.45%) were identified as Staphylococcus aureus. The resistance profile of these isolates showed high resistance to different antibiotics, with 100% of the isolates resistant to chloramphenicol, clindamycin, and erythromycin, 98.33% resistant to doxycycline, 95% resistant to oxacillin, and 85% resistant to cefoxitin. Regarding the mecA gene, 27 (45%) samples were positive for the presence of this gene, and three (11.11%) were phenotypically sensitive to oxacillin and cefoxitin. This finding highlights the importance of researching the phenotypic profile of resistance to different antimicrobials and resistance genes in the different phases of pig rearing to identify the real risk of these isolates from a One Health perspective. The present study revealed the presence of samples resistant to different antibiotics in recently weaned production animal that had not been markedly exposed to antimicrobials as growth promoters or even as prophylactics. This information highlights the need for more research on the possible sharing of bacteria between sows and piglets, the environmental pressure within production environments, and the exposure of handlers during their transport, especially considering the community, hospital, and political importance of the presence of circulating resistant strains.
Assuntos
Infecções Estafilocócicas , Doenças dos Suínos , Animais , Suínos , Feminino , Cefoxitina , Coagulase , Cavidade Nasal/química , Proteínas de Bactérias/genética , Proteínas de Ligação às Penicilinas/genética , Testes de Sensibilidade Microbiana/veterinária , Antibacterianos/farmacologia , Oxacilina , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/genética , Staphylococcus/genética , Doenças dos Suínos/microbiologiaRESUMO
The membrane of methicillin-resistant Staphylococcus aureus (MRSA) contains penicillin-binding proteins (PBPs) in the phospholipidic bilayer, with the protein PBP2a being linked with the resistance mechanism. In this work we confirm the role of PBP2a with molecular-level information obtained with Langmuir monolayers as cell membrane models. The MRSA cell membrane was mimicked with a mixed monolayer of dipalmitoyl phosphatidyl glycerol (DPPG) and cardiolipin (CL), also incorporating PBP2a. The surface pressure-area isotherms and the Brewster angle microscopy (BAM) images for these mixed monolayers were significantly affected by the antibiotic meropenem, which is PBP2a inhibitor. The meropenem effects were associated with the presence of PBP2a, as they were absent in the Langmuir monolayers without PBP2a. The relevance of PBP2a was confirmed with results where the antibiotic methicillin, known to be unsuitable to kill MRSA, had the same effects on mixed DPPG/CL and DPPG/CL-PBP2a monolayers since it prevented PBP2a from incorporating in the monolayer. The biological implication of the findings presented here is that a successful antibiotic against MRSA should be able to interact with PBP2a, but in the membrane.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Meropeném/metabolismo , Meropeném/farmacologia , Meticilina/farmacologia , Testes de Sensibilidade Microbiana , Proteínas de Ligação às Penicilinas/metabolismo , Proteínas de Ligação às Penicilinas/farmacologiaRESUMO
Abatsract Pseudomonas aeruginosa is an important nosocomial pathogen and its clinical importance is mainly related to nosocomial infections. Increased rates of bacterial resistance in recent years has led WHO to publish a global priority list to guide research and discovery of new antibiotics, where P. aeruginosa is among the group of bacteria for which there is a critical level of priority for new drugs to be discovered. In this context, isoeugenol appears as an interesting alternative and the objective of this study was to investigate its action against P. aeruginosa. Isoeugenol presented significant antibacterial activity, with minimum inhibitory concentration (MIC) of 64µg/mL and minimum bactericidal concentration (MBC) of 128µg/mL, and was considered bactericidal against this species. Molecular docking revealed interactions that suggest that isoeugenol may bind to the enzyme Penicillin-Binding Protein 3 and interfere with the bacterial cell wall synthesis process. This study reinforces the antibacterial potential of this compound and emphasizes that more studies are needed in order to better investigate its mechanism of antibacterial action.
Assuntos
Pseudomonas aeruginosa/efeitos dos fármacos , Antibacterianos/efeitos adversos , Bactérias/classificação , Organização Mundial da Saúde , Testes de Sensibilidade Microbiana/instrumentação , Proteínas de Ligação às Penicilinas/agonistas , Medicamentos de Referência , Simulação de Acoplamento Molecular/métodosRESUMO
Infections caused by Staphylococcus aureus are increasingly prevalent, and treatment has become more difficult due to the emergence of strains that are resistant to multiple drugs, such as methicillin-resistant Staphylococcus aureus (MRSA). Penicillin-binding proteins (PBPs) are essential enzymes in peptidoglycan biosynthesis. Only found in bacteria, they are an excellent target for the development of bacterial control strategies. S. aureus has 4 PBPs, and only PBP2 has transglycosylation activity, making it a good model to evaluate whether the inactivation of the transglycosylase domain (PBP2t) could lead to bacterial death. (His6)-tagged PBP2t was purified from the E. coli cell lysate using Ni-charged resin, and ELISA and immunoblotting assays demonstrated that PBP2t is immunogenic. Flow cytometry analysis was performed to verify the binding of polyclonal antibodies to the bacterial cell surface. In order to verify the ability to provide protection, immunized mice were challenged with a sublethal dose of MRSA, and the bacterial loads in kidneys and spleen were evaluated. A reduction of 2-2.5 logs was seen in organs from immunized mice compared with the negative controls in two independent assays (p < 0.01). Our results demonstrate that the PBP2t is a promising target for the development of novel antimicrobial strategies, but further testing should be performed to validate the protection conferred by immunization with this protein.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Staphylococcus aureus , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Proteínas de Bactérias , Escherichia coli , Imunoterapia , Camundongos , Proteínas de Ligação às Penicilinas/genéticaRESUMO
Oxacillin-susceptible mecA-positive Staphylococcus aureus (OS-MRSA) isolates have been described worldwide, but data regarding dogs and their owners have not been reported. This study investigated the occurrence of OS-MRSA and MRSA isolates in the nasal mucosa of 241 healthy dogs and 208 owners in the community. S. aureus isolates were characterized by susceptibility testing, detection of the mecA and the Panton-Valentine leukocidin (PVL) genes, staphylococcal chromosome cassette (SCC)mec typing and rep-PCR-RW3A. We report an unprecedented detection of nasal carriage of OS-MRSA in 5.3 % (2/38) of healthy dogs and 1.75 % (1/57) of their owners. We also found MRSA in 2.6 % (1/38) of the dogs and 3.5 % (2/57) of the owners. Only the human isolate was SCCmec IV and PVL-positive. Molecular typing revealed that the same cluster of S. aureus was present in owners and dogs from the same or different families attended at the same veterinary clinic. The three OS-MRSA isolates did not show genetic similarity to each other. Detection of OS-MRSA in this context alerts us to the role of dogs and owners as possible silent reservoirs of this microorganism in the community, which may potentially be misidentified as methicillin-sensitive S. aureus (MSSA) in the laboratory routine, representing an additional threat in antimicrobial therapy for staphylococcal infections.
Assuntos
Doenças do Cão , Infecções Estafilocócicas , Staphylococcus aureus/isolamento & purificação , Animais , Proteínas de Bactérias/genética , Brasil/epidemiologia , Doenças do Cão/epidemiologia , Doenças do Cão/microbiologia , Cães , Humanos , Testes de Sensibilidade Microbiana/veterinária , Oxacilina/farmacologia , Proteínas de Ligação às Penicilinas/genética , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/genéticaRESUMO
Aim: Genomic analysis of a methicillin-resistant Staphylococcus aureus (MRSA) strain cultured from a non-migratory seabird at Fernando de Noronha Archipelago (Brazilian oceanic islands) was carried out to investigate the potential origin of MRSA genetic determinants in an ecological setting with minimal or absent antimicrobial selective pressure, and minimal interaction with humans and domestic animals. Results: The study determined mecA gene homology and the phylogenetic relatedness with mecA described in Staphylococcus sciuri, which was the major Staphylococcus spp. cultured from the birds. Our findings corroborate in silico assumptions that the mecA gene in MRSA strains clinically relevant for humans and animals originates from S. sciuri ancestors. Conclusion: Coagulase-negative staphylococci seem to be natural reservoirs of methicillin-resistant genes to S. aureus, even in environments with very low antimicrobial selection pressure.
Assuntos
Proteínas de Bactérias/genética , Aves/microbiologia , Staphylococcus aureus Resistente à Meticilina/genética , Proteínas de Ligação às Penicilinas/genética , Staphylococcus/genética , Animais , Brasil , Ilhas , Sequenciamento Completo do GenomaRESUMO
Staphylococcus aureus is a versatile and highly adaptable pathogen associated with a wide range of infectious diseases in humans and animals. In the last decades, concern has increased worldwide due to the emergence and spread of methicillin-resistant S. aureus (MRSA) strains shortly after this drug became a therapeutic option. In this study, we report the genomic features of the first mecC-mediated, ß-lactam resistant MRSA strain associated with livestock in Brazil and in the American continent. Three clonally related phenotypic MRSA isolates originated from a dairy herd were confirmed by polymerase chain reaction as mecC-harbouring MRSA isolates. Whole-genome sequencing was performed by Illumina Miseq platform. Downstream analyses showed that the strain was identified as the sequence type 126 (ST126) and spa type t605. In silico analysis revealed a mecC homolog gene in the orfX region associated with different penicillin-binding proteins. Moreover, genes encoding for efflux pump systems (arlR, mepR, LmrS, norA and mgrA), and antibiotic inactivation enzymes (blaZ and FosB) were also detected. Virulence analyses revealed that the strain harbours genes encoding for exoenzymes (aur, splA, splB and splE), toxin (hlgA, hlgB, hlgC, lukD and lukE) and enterotoxin (sea). The epidemiologic and genomic information provided by this study will support further epidemiological and evolutionary investigations to understand the origin and dissemination of mecC-MRSA among animals and its impact on public health.