Reducing bioassay variability by identifying sources of variation and controlling key parameters in assay protocol

Reducing bioassay variability by identifying sources of variation and controlling important parameters in assay protocol was demonstrated in this study. The variability of a bioassay based on a luminescent bacterium was examined as an example. This assay involved the growth of cells, storage at a low temperature, activation, and exposure to a test sample, and the assay response was bioluminescence inhibition. After determining that measurement error was small and negligible, the total assay variability was decomposed in an initial variance components study into between-batch, between-vial, and between-tube variations. Results indicated that between-vial variations accounted for the majority of the total observed variability and that reducing this type of variation would be beneficial. Five parameters in the assay protocol were determined as factors that potentially affected assay variability significantly. A split-plot design was employed to investigate the effects of these factors and some of their interactions on the assay response. One of the five factors, i.e., activation temperature, turned out to have a significant effect. The variance components study was repeated with better control of activation temperature as well as other parameters. Results indicated that the total variability of the bioassay was reduced by approximately 85%.     

Solute transport in crystalline rocks at Aspö--I: geological basis and model calibration

Water-conducting faults and fractures were studied in the granite-hosted Asp? Hard Rock Laboratory (SE Sweden). On a scale of decametres and larger, steeply dipping faults dominate and contain a variety of different fault rocks (mylonites, cataclasites, fault gouges). On a smaller scale, somewhat less regular fracture patterns were found. Conceptual models of the fault and fracture geometries and of the properties of rock types adjacent to fractures were derived and used as input for the modelling of in situ dipole tracer tests that were conducted in the framework of the Tracer Retention Understanding Experiment (TRUE-1) on a scale of metres. After the identification of all relevant transport and retardation processes, blind predictions of the breakthroughs of conservative to moderately sorbing tracers were calculated and then compared with the experimental data. This paper provides the geological basis and model calibration, while the predictive and inverse modelling work is the topic of the companion paper [J. Contam. Hydrol. 61 (2003) 175]. The TRUE-1 experimental volume is highly fractured and contains the same types of fault rocks and alterations as on the decametric scale. The experimental flow field was modelled on the basis of a 2D-streamtube formalism with an underlying homogeneous and isotropic transmissivity field. Tracer transport was modelled using the dual porosity medium approach, which is linked to the flow model by the flow porosity. Given the substantial pumping rates in the extraction borehole, the transport domain has a maximum width of a few centimetres only. It is concluded that both the uncertainty with regard to the length of individual fractures and the detailed geometry of the network along the flowpath between injection and extraction boreholes are not critical because flow is largely one-dimensional, whether through a single fracture or a network. Process identification and model calibration were based on a single uranine breakthrough (test PDT3), which clearly showed that matrix diffusion had to be included in the model even over the short experimental time scales, evidenced by a characteristic shape of the trailing edge of the breakthrough curve. Using the geological information and therefore considering limited matrix diffusion into a thin fault gouge horizon resulted in a good fit to the experiment. On the other hand, fresh granite was found not to interact noticeably with the tracers over the time scales of the experiments. While fracture-filling gouge materials are very efficient in retarding tracers over short periods of time (hours-days), their volume is very small and, with time progressing, retardation will be dominated by altered wall rock and, finally, by fresh granite. In such rocks, both porosity (and therefore the effective diffusion coefficient) and sorption K(d)s are more than one order of magnitude smaller compared to fault gouge, thus indicating that long-term retardation is expected to occur but to be less pronounced.     

Earthworm biomass as additional information for risk assessment of heavy metal biomagnification: a case study for dredged sediment-derived soils and polluted floodplain soils

The important role of earthworms in the biomagnification of heavy metals in terrestrial ecosystems is widely recognised. Differences in earthworm biomass between sites is mostly not accounted for in ecological risk assessment. These differences may be large depending on soil properties and pollution status. A survey of earthworm biomass and colonisation rate was carried out on dredged sediment-derived soils (DSDS). Results were compared with observations for the surrounding alluvial plains. Mainly grain size distribution and time since disposal determined earthworm biomass on DSDS, while soil pollution status of the DSDS was of lesser importance. Highest earthworm biomass was observed on sandy loam DSDS disposed at least 40 years ago.     

Analytical solutions to compartmental indoor air quality models with application to environmental tobacco smoke concentrations measured in a house

This paper derives the analytical solutions to multi-compartment indoor air quality models for predicting indoor air pollutant concentrations in the home and evaluates the solutions using experimental measurements in the rooms of a single-story residence. The model uses Laplace transform methods to solve the mass balance equations for two interconnected compartments, obtaining analytical solutions that can be applied without a computer. Environmental tobacco smoke (ETS) sources such as the cigarette typically emit pollutants for relatively short times (7-11 min) and are represented mathematically by a "rectangular" source emission time function, or approximated by a short-duration source called an "impulse" time function. Other time-varying indoor sources also can be represented by Laplace transforms. The two-compartment model is more complicated than the single-compartment model and has more parameters, including the cigarette or combustion source emission rate as a function of time, room volumes, compartmental air change rates, and interzonal air flow factors expressed as dimensionless ratios. This paper provides analytical solutions for the impulse, step (Heaviside), and rectangular source emission time functions. It evaluates the indoor model in an unoccupied two-bedroom home using cigars and cigarettes as sources with continuous measurements of carbon monoxide (CO), respirable suspended particles (RSP), and particulate polycyclic aromatic hydrocarbons (PPAH). Fine particle mass concentrations (RSP or PM3.5) are measured using real-time monitors. In our experiments, simultaneous measurements of concentrations at three heights in a bedroom confirm an important assumption of the model-spatial uniformity of mixing. The parameter values of the two-compartment model were obtained using a "grid search" optimization method, and the predicted solutions agreed well with the measured concentration time series in the rooms of the home. The door and window positions in each room had considerable effect on the pollutant concentrations observed in the home. Because of the small volumes and low air change rates of most homes, indoor pollutant concentrations from smoking activity in a home can be very high and can persist at measurable levels indoors for many hours.     

Effects of a mixture of tetracyclines to Lemna gibba and Myriophyllum sibiricum evaluated in aquatic microcosms

The impact of a mixture of oxytetracycline, chlortetracycline, tetracycline and doxycycline on Myriophyllum sibiricum and Lemna gibba was investigated using fifteen 12,000-L microcosms (k=5, n=3). Significant concentration-response relationships were only found for M. sibiricum, where dry mass was 69, 47, 30, and 7% of controls at respective treatment concentrations of 0.080, 0.218, 0.668, and 2.289 micromol/L. Somatic endpoints were strongly and negatively correlated with percent light transmission, except plant length, which was positively correlated. Treated microcosms experienced a reduction in the percent of surface irradiance penetrating the water column as high as 99.8% at a depth of 70 cm, relative to controls. Position relative to the water column was likely responsible for the differential effects observed between floating (L. gibba) and submerged (M. sibiricum) species of macrophytes. A hazard quotient assessment of the lowest EC10 value indicated significant risk, exceeding the critical HQ value, but not the lowest EC25 value.     

Evidence for destruction of PCBs by the OH radical in urban atmospheres

Evidence for reaction of polychlorinated biphenyl (PCB) congeners with the hydroxyl (OH) radical in the troposphere was observed in diurnal variations in ambient gas-phase PCB concentrations at three urban sampling sites located in the Chicago, IL; Baltimore, MD; and Jersey City, NJ urban/industrial areas. The magnitude of the depletion of individual PCB congeners decreased by about 10-20% for each additional chlorine substituent, reflecting slower reaction rates for higher MW congeners with the OH radical. Octa- and nonachlorobiphenyls, which are largely unreactive with the OH radical, were used as tracers to investigate the effects of dilution on diurnal variation. The environmental rate constants for disappearance of the PCBs range from about 1.0 day(-1) for trichlorobiphenyls to about 0.3 day(-1) for hexachlorobiphenyls. Assuming a OH radical concentration of 3 x 10(6) molecules cm (-3), the second-order rate constants for reaction of specific congeners with the OH radical are consistent with laboratory measurements. More importantly, the relative reactivity of PCB homologues agrees well with the relationship predicted by other researchers from laboratory measurements, suggesting that losses of PCBs during daytime tropospheric transport are due at least in part to reactions with the OH radical.     

Incineration and human health

After pollutants from an incineration facility disperse into the air, some people close to the facility may be exposed directly through inhalation or indirectly through consumption of food or water contaminated by deposition of the pollutants from air to soil, vegetation, and water. For metals and other pollutants that are very persistent in the environment, the potential effects may extend well beyond the area close to the incinerator. Persistent pollutants can be carried long distances from their emission sources, go through various chemical and physical transformations, and pass numerous times through soil, water, or food.     

Control of microbial growth and of amphipod grazing by water-soluble compounds from leaves of Zostera marina

The role of minor components of the leaves of Zostera marina L. in altering the activity of micro-organisms directly (and indirectly by affecting amphipod grazers) was investigated in laboratory experiments, using plants collected at Roberts Bank (49°2N; 123°8W) on the west coast of Canada. Water-soluble extracts of eelgrass leaves inhibited the growth of a micro-alga (Platymonas sp.) and many marine bacteria at concentrations equivalent to as little as 10 mg dry leaf l^-1. The potency of leaf extracts was higher (1) in young, actively metabolizing tissue than in older leaves, and (2) in leaves collected during rapid growth in summer 1980 than during the following winter. Water-soluble inhibitors (especially phenolic acids) may explain the low biomass of epiphytes on actively growing leaves. Three phenolic acids inhibited the test micro-organisms at concentrations as low as 0.3 mg l^-1; caffeic acid was more potent than either protocatechuic or gentisic acid. Extracts of young leaves also inhibited grazing by amphipods [Eogammarus confervicolus (Stimpson)] on dead leaves. The time required for leaching of soluble inhibitors may account for the delay between the loss of leaves from the plants and the onset of grazing. Thus, interactions among the biotic components of the detritus ecosystem may be significantly modified by minor compounds in the leaves of Z. marina