RESUMO
Staphylococcus lugdunensis produces lugdulysin, a metalloprotease that may contribute to its virulence. This study aimed to evaluate the biochemical aspects of lugdulysin and investigate its effect on Staphylococcus aureus biofilms. The protease was isolated and characterized for its optimal pH and temperature, hydrolysis kinetics, and influence of metal cofactor supplementation. The protein structure was determined via homology modeling. The effect on S. aureus biofilms was assessed by the micromethod technique. The protease optimal pH and temperature were 7.0 and 37 °C, respectively. EDTA inhibited protease activity, confirming it as a metalloprotease. Lugdulysin activity was not recovered by divalent ion supplementation post-inhibition, and supplementation with divalent ions did not change enzymatic activity. The isolated enzyme was stable for up to 3 h. Lugdulysin significantly inhibited the formation and disrupted preestablished protein-matrix MRSA biofilm. This preliminary study indicates that lugdulysin has a potential role as a competition mechanism and/or modulation of staphylococcal biofilm.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Staphylococcus lugdunensis , Humanos , Staphylococcus aureus , Antibacterianos/farmacologia , Biofilmes , Metaloproteases/farmacologia , Peptídeo Hidrolases , Testes de Sensibilidade MicrobianaRESUMO
INTRODUCCIÓN: Staphylococcus lugdunensis, es un estafilococo coagulasa negativa (SCN) con características de virulencia y de sensibilidad antimicrobiana que lo hacen más parecido a Staphylococcus aureus que a otros SCN. OBJETIVOS: Conocer las características clínicomicrobiológicas de los aislados de S. lugdunensis identificados en nuestra institución. MATERIAL Y MÉTODOS: Se realizó un estudio retrospectivo de los aislados de S. lugdunensis entre los años 2017 y 2019 en el Servicio de Microbiología del Hospital Universitario San Jorge de Huesca (España). Se revisaron las historias clínicas correspondientes a los pacientes con aislamiento de S. lugdunensis, considerándose las siguientes variables: edad, sexo, tipo de muestra, servicio de procedencia y enfermedad de base. La identificación bacteriana se realizó con MALDI-TOF VITEK MS (BioMérieux, Francia). Así mismo, se estudió su patrón de susceptibilidad antimicrobiana in vitro mediante microdilución en placa. RESULTADOS: Se obtuvieron 44 aislados de S. lugdunensis: 12 procedían de heridas, 10 fueron abscesos, 8 úlceras, 7 orinas, 4 frotis cutáneos, 2 exudados óticos, y 1 exudado vaginal. En relación con la enfermedad de base destacaron cinco pacientes con procesos tumorales y diez con diabetes mellitus. En 17 pacientes existían antecedentes de cirugía o traumatismo reciente. La mayoría de las cepas fueron sensibles a los antimicrobianos estudiados. En 19 de ellas se observó producción de β-lactamasa, dos fueron resistentes a macrólidos y tres a clindamicina. Todas las cepas fueron sensibles a oxacilina, gentamicina y cotrimoxazol. CONCLUSIONES: Aunque S. lugdunensis mantiene una buena sensibilidad a la mayoría de los antimicrobianos, su tendencia a producir abscesos y que exprese factores de virulencia más parecido a S. aureus que a otros SCN, hace necesaria una correcta identificación en el laboratorio con el fin de que su incidencia no quede subestimada.
BACKGROUND: Staphylococcus lugdunensis is a coagulase-negative staphylococcus (CNS) with virulence and antibiotic sensitivity characteristics which makes it more similar to Staphylococcus aureus than other CNS. AIM: To know the microbiological and clinical characteristics of S. lugdunensis isolates identified from our health sector. METHODS: A retrospective study of S. lugdunensis isolates was carried out between 2017 and 2019 in the Microbiology Service of the San Jorge University Hospital in Huesca (Spain). The clinical records of patients with S. lugdunensis isolation were reviewed, considering the following factors: age, sex, sample type, service and underlying disease. Bacterial identification was performed using MALDI-TOF VITEK MS (BioMérieux, France). The pattern of antibiotic susceptibility was studied by means of plate microdilution. RESULTS: 44 isolates of S. lugdunensis were obtained: 12 corresponded to wounds, 10 were abscesses, 8 ulcers, 7 urine samples, 4 skin smears, 2 otic exudates, and 1 vaginal exudate. Regarding the underlying disease, five patients had a tumor processes and ten had diabetes mellitus. In 17 patients there was a history of recent surgery or trauma. Most of the strains were susceptible to the antibiotics studied. Production of beta-lactamase was observed in 19 of them, two were resistant to macrolides and three to clindamycin. None of the isolates were resistant to oxacillin, gentamicin or cotrimoxazole. CONCLUSIONS: Although S. lugdunensis maintains a good sensitivity to most antibiotics, its tendency to produce abscesses and that it expresses virulence factors more similar to S. aureus than to other CNS requires a correct identification in the laboratory so that its incidence is not underestimated.
Assuntos
Humanos , Masculino , Feminino , Lactente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Infecções Estafilocócicas/microbiologia , Staphylococcus lugdunensis , Oxacilina , Staphylococcus aureus , beta-Lactamases , Clindamicina , Gentamicinas , Testes de Sensibilidade Microbiana , Combinação Trimetoprima e Sulfametoxazol , Estudos Retrospectivos , Coagulase , Macrolídeos , Fatores de Virulência , Abscesso/tratamento farmacológico , Antibacterianos/uso terapêutico , Antibacterianos/farmacologiaRESUMO
El Staphylococcus lugdunensis se manifiesta más como Staphylococcus aureus que como un típico estafilococo coagulasa negativa, especialmente por su gran virulencia y capacidad para producir infecciones supuradas. Debido al protagonismo creciente de esta especie, es propósito de esta investigación destacar la significación clínica y los aspectos microbiológicos para el diagnóstico de pacientes con dicha infección.
The Staphylococcus lugdunensis is manifested more as Staphylococcus aureus that as a typical negative coagulase staphylococcus, especially for its great virulence and capacity to produce suppurated infections. Due to the growing protagonism of this species, this investigation aims at highlighting the clinical significance and microbiological aspects for the diagnosis of patients with this infection.
Assuntos
Técnicas e Procedimentos Diagnósticos , Staphylococcus lugdunensis , Staphylococcus aureus , Staphylococcus lugdunensis/isolamento & purificaçãoRESUMO
Staphylococcus lugdunensis is a coagulase-negative staphylococcus of growing importance and atypical behavior. The infections caused by this microorganism are becoming more frequent, having a broader spectrum. Psoas abscesses caused by this germ are rare, with few cases reported in the literature. In this work, we present a case of a psoas abscess caused by S. lugdunensis in a patient suffering from diabetes mellitus and rheumatoid arthritis, which was treated with intravenous cloxacillin with a good outcome.
Assuntos
Abscesso do Psoas/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus lugdunensis/isolamento & purificação , Antibacterianos/uso terapêutico , Artrite Reumatoide/complicações , Técnicas de Tipagem Bacteriana , Cloxacilina/uso terapêutico , Diabetes Mellitus Tipo 2/complicações , Abscesso Epidural/tratamento farmacológico , Abscesso Epidural/microbiologia , Feminino , Humanos , Hospedeiro Imunocomprometido , Pessoa de Meia-Idade , Abscesso do Psoas/tratamento farmacológico , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus lugdunensis/patogenicidadeRESUMO
Os estafilococos coagulase negativos (ECNs) são cocos Gram-positivos usualmente considerados contaminantes em laboratórios de microbiologia clínica. Apesar de pertencer a este grupo, Staphylococcus lugdunensis pode causar infecções complicadas, como endocardites, infecções de pele e tecidos moles, osteomielites, entre outras. Além da formação de biofilmes, apresenta patogenicidade similar ao Staphylococcus aureus. É um dos principais agentes causadores de endocardites, com taxa de mortalidade de até 70 por cento. Pode ser confundido com S. aureus quando se utilizam testes rápidos para sua identificação, como a pesquisa de clumping factor, no caso de teste de coagulase em lâmina, ou em testes de aglutinação direta em látex. Pode ser facilmente identificado por meio de provas bioquímicas acessíveis, como a presença de atividade da ornitina descarboxilase e pirrolidonil arilamidase (PYR). Apresenta sensibilidade à maioria dos agentes antimicrobianos, devendo ser pesquisada rotineiramente a presença de betalactamases e do gene mecA por meio de testes com cefalosporina cromogênica e suscetibilidade à cefoxitina, respectivamente. Convém salientar que os critérios interpretativos utilizados para avaliar a sensibilidade à cefoxitina são os mesmos preconizados para S. aureus e diferentes dos utilizados para os outros ECNs. Apesar de incomum, o S. lugdunensis é um patógeno com acentuada virulência que deve ser corretamente identificado, pois raramente poderá ser considerado contaminante quando isolado de sítios estéreis.
Coagulase-negatives staphylococci (CNS) are Gram-positives cocci commonly regarded as contaminants in clinical microbiology laboratories. Despite belonging to this group, Staphylococcus lugdunensis may cause complicated infections such as endocarditis, skin infections and soft tissue, osteomyelitis, among others. Apart from the formation of biofilms, it has pathogenic features similar to Staphylococcus aureus. It may be mistakenly identified as S. aureus when using rapid identification tests, such as clumping factor in slide coagulase or in agglutination latex tests. It is easily identified through available biochemical tests, such as the presence of ornithine decarboxylase and pyrrolidonyl arylamidase (PYR). It presents sensitivity to most antimicrobial agents. Furthermore, the presence of beta-lactamase and mecA gene should be routinely investigated by testing with chromogenic cephalosporin and cefoxitin susceptibility, respectively. It is convenient to highlight that the interpretative criteria used to evaluate cefoxitin sensitivity are the same recommended for S. aureus and different from those used for other CNS. Despite the fact it is atypical, S. lugdunensis is a virulent pathogen, which must be accurately identified insofar as it will rarely be deemed as a contaminant when isolated from sterile sites.