Optimal culture incubation time in orthopedic device-associated infections: a retrospective analysis of prolonged 14-day incubation.

Accurate diagnosis of orthopedic device-associated infections can be challenging. Culture of tissue biopsy specimens is often considered the gold standard; however, there is currently no consensus on the ideal incubation time for specimens. The aim of our study was to assess the yield of a 14-day incubation protocol for tissue biopsy specimens from revision surgery (joint replacements and internal fixation devices) in a general orthopedic and trauma surgery setting. Medical records were reviewed retrospectively in order to identify cases of infection according to predefined diagnostic criteria. From August 2009 to March 2012, 499 tissue biopsy specimens were sampled from 117 cases. In 70 cases (59.8%), at least one sample showed microbiological growth. Among them, 58 cases (82.9%) were considered infections and 12 cases (17.1%) were classified as contaminations. The median time to positivity in the cases of infection was 1 day (range, 1 to 10 days), compared to 6 days (range, 1 to 11 days) in the cases of contamination (P < 0.001). Fifty-six (96.6%) of the infection cases were diagnosed within 7 days of incubation. In conclusion, the results of our study show that the incubation of tissue biopsy specimens beyond 7 days is not productive in a general orthopedic and trauma surgery setting. Prolonged 14-day incubation might be of interest in particular situations, however, in which the prevalence of slow-growing microorganisms and anaerobes is higher.

Optimization of incubation time of protein Tc85 in the construction of biosensor: Is the EIS a good tool?

Electrochemical impedance spectroscopy (EIS) in pH 6.9 phosphate buffer solution was used to investigate each step of the procedure employed to modify a screen-printed electrode (SPE). The SPE was modified with self-assembled monolayers (SAMs) of cystamine (CYS, deposited from 20 mM solution), followed by glutaraldehyde (GA, 0.3 M solution). The Trypanosoma cruzi antigen was immobilized using different deposition times. The influence of incubation time (2-18 h) of protein was also investigated. The topography of modified electrode with this protein was investigated by atomic force microscopy (AFM). Interpretation of impedance data was based on physical and chemical adsorption, and degradation of the layer at high and meddle frequencies, and charge transfer reaction involving mainly the reduction of oxygen at low frequencies. EIS studies on modified electrodes with Tc85 protein immobilized for different incubation times indicated that the optimum incubation time was 6-8 h. It was demonstrated that EIS is a good technique to evaluate the different steps and the integrity of the surface modifications, and to optimize the incubation time of protein in the development of biosensors. (C) 2010 Elsevier B.V. All rights reserved.

Optimization of incubation time of protein Tc85 in the construction of biosensor: Is the EIS a good tool?

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bovine sperm cells viability during incubation with or without exogenous DNA

The aim of this study was to assess the effect of exogenous DNA and incubation time on the viability of bovine sperm. Sperm were incubated at a concentration of 5 x 10(6)/ml with or without plasmid pEYFP-NUC. Fluorescent probes, propidium iodide/Hoechst 33342, FITC-PSA and JC-1, were used to assess plasma membrane integrity (PMI), acrosome membrane integrity (AMI) and mitochondrial membrane potential (MMP) respectively at 0, 1, 2, 3 and 4 h of incubation. Exogenous DNA addition did not affect sperm viability; however, incubation time was related to sperm deterioration. Simultaneous assessment of PMI, AMI and MMP showed a reduction in the number of sperm with higher viability (integrity of plasma and acrosome membranes and high mitochondrial membrane potential) from 58.7% at 0 h to 7.5% after 4 h of incubation. Lower viability sperm (damaged plasma and acrosome membranes and low mitochondrial membrane potential) increased from 4.6% at 0 h to 25.99% after 4 h of incubation. When PMI, AMI and MMP were assessed separately we noticed a reduction in plasma and acrosome membrane integrity and mitochondrial membrane potential throughout the incubation period. Therefore, exogenous DNA addition does not affect sperm viability, but the viability is reduced by incubation time.

Effect of time-temperature binomial in obtaining biochemical oxygen demand of different wastewaters

Studies on the effects of temperature and time of incubation of wastewater samples for the estimation of biodegradable organic matter through the biochemical oxygen demand (BOD), that nowadays are rare, considering that the results of the classic study of STREETER & PHELPS(1925) have been accepted as standard. However, there are still questions how could be possible to reduce the incubation time; whether the coefficient of temperature (θ) varies with the temperature and with the type of wastewater and if it approaches 1.047. Aiming the elucidation of these questions, wastewater samples of dairy, swine and sewage treated in septic tanks were incubated at temperatures of 20, 30 and 35 °C, respectively for 5, 3.16 and 2.5 days. From the parameter of deoxygenation coefficient at 20 °C (k20), θ30 and θ35 were calculated. The results indicated that θ values changes with the type of wastewater, however does not vary in the temperature range between 30 and 35 °C, and that the use of 1.047 value did not implied significant differences in obtaining k in a determined T temperature. Thus, it is observed that the value of θ can be used to estimate the required incubation time of the samples at different temperatures.

Effects of Incubation Temperature and Relative Humidity on Embryonic Development in Eggs of Red-Winged Tinamou (Rhynchotus rufescens).

This study was conducted to assess the effects of incubation temperature (34 C, 36[degree]C and 38[degree]C) and relative humidity (RH, 50% and 60%) on egg weight loss, embryo mortality, hatchability, incubation time and chick weight in eggs from red-winged tinamou. The eggs were placed in incubators that were operated at 34[degree]C, 36[degree]C, or 38[degree]C and 50% or 60% RH (mean wet bulb temperatures of 28[degree]C and 30[degree]C, respectively) from day 1 to hatching. Each treatment had two replicate groups of 30 eggs each. Hatchability varied with incubation temperature and RH and was highest for eggs incubated at 36[degree]C and 60% RH and lowest for eggs incubated at 38[degree]C. Early, intermediate and late embryo mortality were highest at 38[degree]C, 38[degree]C/50% RH, and 50% RH, respectively. Incubation period was longest at 34[degree]C and shortest at 38[degree]C/50% RH. Present results show the highest hatchability of red-winged tinamou eggs after incubation at 36[degree]C and 60% RH; highest embryo sensitivity to high temperature in the early period of incubation (1 to 7 days), to high temperature and low RH in the second period of incubation (8-14 days) and to low RH in the late period of incubation (after 15 days) and shortest incubation period with increasing temperature and RH.

Dissolved lithium and silicon through time in microbial dissolution experiments on ODP Leg 209 peridotite

Using peridotite drilled during Ocean Drilling Program Leg 209, a series of enrichment cultures were initiated on board the ship to stimulate microbially enhanced dissolution of olivine. Dissolution was estimated by measured changes in dissolved Li and Si in the media through time (up to 709 days). The results suggest that there was no significant difference between the amounts of dissolved Li and Si in most of the inoculated microbial cultures compared to the control cultures. Alternative explanations for this are that 1. No microbes are living in the culture tubes that can affect the dissolution rates of olivine, 2. The control cultures have microbes effecting the dissolution of olivine as well as the inoculated cultures, 3. Not enough time has passed to build up a large enough microbial population to effect the dissolution of the olivine in the culture tubes, 4. Microbes act to suppress dissolution of olivine instead of enhancing dissolution, and 5. Abiotic dissolution overshadows microbially enhanced dissolution. Further work is required to test these alternatives.