The main goal of this study is to alert researchers who work with cell cultures for the risk of contamination by structures called nanobacteria (NB). NB are tiny structures with size varying from 80 to 500 nm, commonly occurring in clusters and producing a biofilm which contains carbonate or hydroxyl apatite. The most likely source of cell culture contamination by such organisms is serum used as supplement in culture media. The presence of NB leads to a progressive culture deterioration with accumulation of granules (probably phagocytized NB) in cytoplasmic vacuoles, an increasing number of dead cells in the supernatant and degeneration of cells that remained attached to the bottom of the vessel. NB can also be found in culture supernatants where they are found in clusters with variable size and displaying brownian movement. In this study, 19 cell lineages, 8 batches of sera and 1 batch of growth supplement from different sources were analyzed. Samples from sera were cultured in Eagle's Minimum Essential Medium (E-MEM) or incubated directly at 37ºC. Tests carried out to detect the presence of extracellular bacteria, Mycoplasma sp and viruses were all negative. Analysis by scanning electron microscopy (SEM) revealed tiny oval structures less than 500 nm in size, isolated or in small groups, in all material analyzed except in one fetal bovine serum batch.
Storage tanks containing microbially contaminated diesel oil are susceptible to corrosion. This process may be evaluated electrochemically in the laboratory using simulated storage systems containing diesel oil and an aqueous phase. The simulated aqueous phase must supply mineral nutrients for microbial growth, together with adequate electrical conductivity, without, however, being too corrosive, so as to allow the aggressive nature of the microbial metabolites to be detected. In this investigation, microbial growth was measured in six electrically conductive media overlaid with metropolitan diesel oil containing an additive package. The microorganisms were the filamentous fungi, Hormoconis resinae, Paecilomyces variotii and Aspergillus fumigatus, the bacterium Bacillus subtilis and the yeast Candida silvicola, all previously isolated from contaminated diesel oil. After 60 days incubation with pure or mixed inocula of these microorganisms, pH, conductivity and viable microorganisms were measured. The electrochemical behaviour of carbon steel ASTM 283-93-C was determined in each of the six media (uninoculated) and in selected inoculated medium via measurements of open circuit potential and potentiostatic polarization curves. The uptake of phosphate (corrosion inhibitor), microbial growth, pH, conductivity and anodic and cathodic polarization curves were assessed in the water phase after 30 and 60 days of incubation with each single species Aspergillus fumigatus and Hormoconis resinae and with the consortium. The medium which proved most appropriate was Bushnell-Haas medium modified by the omission of chlorides, which allowed satisfactory microbial growth and had low aggressivity towards the steel. The performance of electrochemical tests in aerated, rather than deaerated, electrolyte solutions is suggested to be important to allow the detection of microbial influence on passive film formation and stability.
Objetivos: Verificar a aderência bacteriana a lentes intra-oculares de silicone e de polimetilmetacrilato como possível fator de risco no desenvolvimento de endoftalmite pós-operatória, utilizando-se um modelo in vitro com três microrganismos potencialmente patogênicos. Métodos: As análises foram realizadas com cepas de Staphylococcus aureus ATCC 29213, Staphylococcus epidermidis (amostra clínica) e Pseudomonas aeruginosa ATCC 27853 incluindo a determinação de curvas de crescimento, testes para verificação de produção de cápsula, avaliação da hidrofobicidade, testes de aderência a diferentes materiais, microscopia óptica, microscopia eletrônica de varredura e microscopia de força atômica. Resultados: A produção de cápsula e a aderência das três diferentes cepas não mostraram qualquer relação com a quantidade de microrganismos; em relação às lentes intra-oculares de polimetilmetacrilato e de silicone, não houve diferença estatisticamente significativa na aderência de S. aureus e S. epidermidis; P. aeruginosa foi o microrganismo mais aderente a ambos os materiais. A microscopia eletrônica de varredura confirmou estes achados em relação à aderência, ao peso que a microscopia de força atômica evidenciou a produção de biofilme pelas cepas de S. aureus, S. epidermidis e P. aeruginosa. Conclusões: Constatou-se, in vitro, que os materiais analisados não diferiram com relação à taxa de aderência bacteriana, porém, P. aeruginosa apresentou maior eficiência de adesão entre as bactérias testadas. Todas as cepas produziram biofilme. Silicone foi o material mais hidrofóbico, quando comparado ao polimetilmetacrilato.
INTRODUÇÃOCirurgia de catarata para implante de lentes intra-oculares (LIO) pode levar à endoftalmite pós-operatória e, ainda que raramente, à perda total da visão no olho acometido em até 50% dos casos (1)(2)(3) . A microbiota endógena conjuntival é a primeira suspeita quando se está pesquisando a causa de infecções oculares pós-traumáticas e pós-operatórias (4)(5)(6)(7)(8)(9) . Diferentes tipos de bactérias podem causar endoftalmite sendo estafilococos coagulase-negativos, mais especificamente S. epidermidis, os agentes mais comumente isolados (6,(10)(11)(12)(13) . Sabe-se que S. epidermidis tem habilidade de se aderir e proliferar em superfícies de polímeros, especialmente lentes intra-oculares, secretando uma matriz extracelular viscosa (biofilme) que o protege contra antibióticos, bem como dos mecanismos de defesa do hospedeiro (10,(14)(15)(16) . A adesão bacteriana a lentes intra-oculares e a produção de biofilme parecem
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