Application of a risk management framework to a drinking water supply augmented by stormwater recharge.

The Blue Lake is an important water resource for the city of Mount Gambier and the surrounding region, primarily as the drinking water supply source, but also as a tourist attraction. Mount Gambier's stormwater is discharged directly via drainage wells into the unconfined, karstic Gambier Limestone aquifer, which in turn provides the majority of recharge to Blue Lake. Discharge of urban runoff to the aquifer commenced in the 1800s as a means of stormwater management, but is now recognised as contributing to the drinking water supply in Blue Lake. Recently, guidelines for managing the risks associated with water recycling and augmenting drinking water supplies have been developed. This paper examines the organic chemical hazards associated with a stormwater to potable recycling scheme as an example of the current risk management framework.

Factors affecting the performance and risks to human health of on-site wastewater treatment systems.

Aerobic wastewater treatment systems (aerobic systems) are the preferred choice in a region overlying a karstic aquifer used for drinking water supplies, as they are thought to provide better protection to groundwater and human health than standard septic systems. However, aerobic systems in operation do not always perform to design standard; while this is often blamed on lack of maintenance, few studies have investigated the link directly. This study investigates the performance of domestic on-site wastewater treatment systems in South Australia, and compares effluent quality to maintenance records. Effluent from 29 septic tanks and 31 aerobic systems was analysed for nutrients, physico-chemical parameters and microbiological indicators. Aerobic systems generally provided greater treatment than septic tanks, yet most aerobic systems did not meet regulatory guidelines with high levels of indicator bacteria in 71% of samples. The effect of system size, number of household occupants and maintenance on aerobic system treatment performance was analysed: chlorine levels were positively correlated with time of last service, and nutrient concentrations were positively correlated with the number of occupants. A microbial risk assessment revealed the observed irrigation practices to be high risk; and sufficient residence time in the aquifer cannot be guaranteed for protection of groundwater used for drinking. Additional preventive measures such as irrigation management or post treatment of drinking water supply (such as UV disinfection) are required to meet public health targets.

A cDNA for a protein that interacts with the human immunodeficiency virus Tat transactivator.

The human immunodeficiency virus (HIV) tat protein (Tat) is a positive regulator of virus gene expression and replication. Biotinylated Tat was used as a probe to screen a lambda gt11 fusion protein library, and a complementary DNA encoding a protein that interacts with Tat was cloned. Expression of this protein, designated TBP-1 (for Tat binding protein-1), was observed in a variety of cell lines, with expression being highest in human cells. TBP-1 was localized predominantly in the nucleus, which is consistent with the nuclear localization of Tat. In cotransfection experiments, expression of TBP-1 was able to specifically suppress Tat-mediated transactivation. The strategy described may be useful for direct identification and cloning of genes encoding proteins that associate with other proteins to modulate their activity in a positive or negative fashion.

Isolation of a rel-related human cDNA that potentially encodes the 65-kD subunit of NF-kappa B.

The sequence reported in our 22 March 1991 report "Isolation of a rel-related human cDNA that potentially encodes the 65-kD subunit of NFkappaB" [Science 251, 1490 (1991)], contained some errors. Resequencing under strong denaturing conditions revealed three insertions at nucleotide positions 1194, 1212, and 1220, which changed the AA sequence from RSAR-PRLGP to QISQASALAP (residues 372 to 380), thus accounting for some of the divergence in this region. A corrected sequence has been sent to GenBank.

Isolation of a rel-related human cDNA that potentially encodes the 65-kD subunit of NF-kappa B.

A DNA probe that spanned a domain conserved among the proto-oncogene c-rel, the Drosophila morphogen dorsal, and the p50 DNA binding subunit of NF-kappa B was generated from Jurkat T cell complementary DNA with the polymerase chain reaction (PCR) and degenerate oligonucleotides. This probe was used to identify a rel-related complementary DNA that hybridized to a 2.6-kilobase messenger RNA present in human T and B lymphocytes. In vitro transcription and translation of the complementary DNA resulted in the synthesis of a protein with an apparent molecular size of 65 kilodaltons (kD). The translated protein showed weak DNA binding with a specificity for the kappa B binding motif. This protein-DNA complex comigrated with the complex obtained with the purified human p65 NF-kappa B subunit and binding was inhibited by I kappa B-alpha and -beta proteins. In addition, the 65-kD protein associated with the p50 subunit of NF-kappa B and the kappa B probe to form a complex with the same electrophoretic mobility as the NF-kappa B-DNA complex. Therefore the rel-related 65-kD protein may represent the p65 subunit of the active NF-kappa B transcription factor complex.

Optimizing land management strategies for maximum improvements in lake dissolved oxygen concentrations.

Eutrophication and anoxia are unresolved issues in many large waterbodies. Globally, management success has been inconsistent, highlighting the need to identify approaches which reliably improve water quality. We used a process-based model chain to quantify effectiveness of terrestrial nutrient control measures on in-lake nitrogen, phosphorus, chlorophyll and dissolved oxygen (DO) concentrations in Lake Simcoe, Canada. Across a baseline period of 2010-2016 hydrochemical outputs from catchment models INCA-N and INCA-P were used to drive the lake model PROTECH, which simulated water quality in the three main basins of the lake. Five terrestrial nutrient control strategies were evaluated. Effectiveness differed between catchments, and water quality responses to nutrient load reductions varied between deep and shallow lake basins. Nutrient load reductions were a significant driver of increased DO concentrations, however strategies which reduced tributary inflow had a greater impact on lake restoration, associated with changes in water temperature and chemistry. Importantly, when multiple strategies were implemented simultaneously, resultant large flow reductions induced warming throughout the water column. Negative impacts of lake warming on DO overwhelmed the positive effects of nutrient reduction, and limited the effectiveness of lake restoration strategies. This study indicates that rates of lake recovery may be accelerated through a coordinated management approach, which considers strategy interactions, and the potential for temperature change-induced physical and biological feedbacks. Identified impacts of flow and temperature on rates of lake recovery have implications for management sustainability under a changing climate.

Effects of formulations of the fungicide, pentachloronitrobenzene on early life stage development of the Japanese medaka (Oryzias latipes).

Quintozene is a fungicide containing the active ingredient, pentachloronitrobenzene (PCNB) that is used to control "snow mold" on golf courses in temperate regions of North America. In this study, quintozene and a formulation of quintozene widely used on golf courses, FFII were tested for toxicity to early life stages of the Japanese medaka, Oryzias latipes. For medaka exposed in static non-renewal assays to quintozene for 17d from the fertilized egg stage to yolk resorption at the fry stage, the LC(50) for mortality was a nominal concentration of 707 microgl(-1) and the effective concentration for 50% hatch (i.e. EC(50)) was a nominal concentration of 71 microgl(-1). Eggs and fry showed developmental abnormalities, including ocular malformations and retarded development of the brain, notochord, organs and body segmentation, which were interpreted as teratogenic responses to exposure to PCNB. For medaka exposed to quintozene, the LOECs for abnormalities of the eye and all other developmental abnormalities were 750 and 100 microgl(-1), respectively. In medaka exposed to the FFII formulation, similar patterns of mortalities, reduced hatching success and developmental abnormalities were observed, but at higher test concentrations that were consistent with the proportion of quintozene in the formulation. For medaka exposed to the formulation, the LOECs for abnormalities of the eye and all other developmental abnormalities were 10,000 and 1,000microgl(-1), respectively. Overall, these data indicate that studies should be conducted to assess the risk of exposure of early life stages of fish to quintozene in watersheds impacted by golf courses.

Never say never: circumventing a contraindication to control apnoea-induced epileptic events with a mandibular advancement device.

BACKGROUND: The benefit of mandibular advancement devices in patients with sleep-disordered breathing and as a potential option for obstructive sleep apnoea syndrome is well recognised. Their use in the setting of epilepsy or other seizure disorders is typically contraindicated.Case reportA 48-year-old patient with a history of poorly controlled epilepsy and obstructive sleep apnoea syndrome was referred for ENT review for possible tracheostomy. The patient was wheelchair-bound with 24-hour continuous positive airway pressure, but sleep studies demonstrated persistent, severe episodes of apnoea and notable sleep disturbance. Sleep nasendoscopy demonstrated marked improvement on capnography with the laryngeal mask airway in situ, and this was maintained with mandibular advancement using jaw thrust following removal of the laryngeal mask airway. A mandibular advancement device was subsequently trialled; this had no subjective benefit for the patient, but the seizures resolved and control of apnoea was achieved with the combination of a mandibular advancement device and continuous positive airway pressure. CONCLUSION: This paper highlights a novel application of mandibular advancement devices, used in combination with continuous positive airway pressure, which resulted in complete resolution of sleep deprivation and apnoea-induced epileptic events.

Single nucleotide polymorphic discrimination by an electronic dot blot assay on semiconductor microchips.

We have developed a rapid assay for single nucleotide polymorphism (SNP) detection that utilizes electronic circuitry on silicon microchips. The method was validated by the accurate discrimination of blinded DNA samples for the complex quadra-allelic SNP of mannose binding protein. The microchip directed the transport, concentration, and attachment of amplified patient DNA to selected electrodes (test sites) creating an array of DNA samples. Through control of the electric field, the microchip enabled accurate genetic identification of these samples using fluorescently labeled DNA reporter probes. The accuracy of this approach was established by internal controls of dual labeled reporters and by using mismatched sequences in addition to the wild-type and variant reporter sequences to validate the SNP-genotype. The ability to customize this assay for multiple genes has advantages over other existing approaches.

Mercury concentration in black flies Simulium spp. (Diptera, Simuliidae) from soft-water streams in Ontario, Canada.

Total Hg in Simulium spp. (Diptera, Simuliidae) was measured in 17 soft-water streams in the District of Muskoka and Haliburton County (Ontario, Canada) during 2003 and 2004. Black flies contained 0.07-0.64 microg/g total Hg (dry weight). The methylmercury concentration was measured in 6 samples of the 17, and ranged from 58% to 93% of total Hg. The concentration of total Hg is much higher than has been found in other filter feeding insects, and represents a significant potential source of Hg to fish. Mercury concentrations in Simulium spp. at different sites were strongly positively correlated with dissolved organic carbon, and the proportion of land within each catchment that was wetland. There was also a strong negative correlation with pH. By examining Hg concentration in filter feeding insects we have found a significant entry point for Hg and MeHg into the food web.

Climate variability and forecasting surface water recovery from acidification: modelling drought-induced sulphate release from wetlands.

Climate-induced drought events have been shown to have a significant influence on sulphate (SO(4)(2-)) export from forested catchments in central Ontario, subsequently delaying recovery of surface waters from acidification. Field and modelling studies have demonstrated that water table drawdown during drought periods promotes oxidation of previously stored (reduced) sulphur (S) compounds in wetlands, with subsequent efflux of SO(4)(2-) upon re-wetting. Although climate-induced changes in processes are generally not integrated into soil-acidification models, MAGIC (Model of Acidification of Groundwater in Catchments) includes a wetland compartment that incorporates redox processes driven by drought events. The potential confounding influence of climate-induced drought events on acidification recovery at Plastic Lake, south-central Ontario (under proposed future S emission reductions) was investigated using MAGIC and two climate scenarios: monthly precipitation and runoff based on long-term means (average-climate scenario), and variable precipitation and runoff based on the past 20 years of observed monthly data (variable-climate scenario). The variable-climate scenario included several periods of summer drought owing to lower than average rainfall and higher then average temperature. Nonetheless, long-term regional trends in precipitation and temperature suggest that the variable-climate scenario may be a conservative estimate of future climate. The average-climate scenario indicated good recovery potential with acid neutralising capacity (ANC) reaching approximately 40 micromol(c)L(-1) by 2020 and 50 micromol(c)L(-1) by 2080. In contrast, the forecasted recovery potential under the variable-climate scenario was very much reduced. By 2080, ANC was forecasted to increase to 2.6 micromol(c)L(-1) from -10.0 micromol(c)L(-1) in 2000. Elevated SO(4)(2-) efflux following drought events (introduced under the variable-climate scenario) has a dramatic impact on simulated future surface water chemistry. The results clearly demonstrate that prediction of future water quality, using models such as MAGIC, should take into account changes or variability in climate as well as acid deposition.

Structural characterization of broadly neutralizing human monoclonal antibodies against the CD4 binding site of HIV-1 gp120.

The human monoclonal antibodies (mAbs) 15e and 21h are derived from HIV-1-infected individuals. They block CD4 binding, recognize conformation-dependent discontinuous epitopes on gp120 and neutralize a broad range of laboratory strains and primary isolates of HIV-1. To determine if a structural basis for neutralization could be identified, analysis of these CD4-binding site anti-gp120 human mAbs was performed, common features and differences were identified and a comparison was made with F105, a previously reported CD4-binding site anti-gp120 human mAb. The 15e and 21h mAb heavy chains are derived from different V region genes, i.e. V2-1 and VDP-35, which are members of the VHIV and VHIII families, respectively. Analysis of the genes encoding the heavy chain complementarity determining region (CDR) 3 revealed that both mAbs show a long DH segment of similar size that could arise from D-D fusions of the dxp1/dlr1 and daudi/d22-12 germline DH genes along with use of the JH6 and JH5 germline segments. Similarly, the 15e and 21h light chains are derived from different V region genes, i.e. Hum01/012 and Hum1v318, that are members of the V kappa I and V lambda IIIa gene families, respectively. These V genes are rearranged with J kappa 1 and J lambda 2 germline genes. For both mAbs, the pattern of replacement mutations in the V region genes of the heavy and light chains is consistent with a process of somatic mutation and antigen-driven clonal selection. By comparing the CDRs of 15e, 21h and F105, eight positions in the rearranged heavy chains and two positions in the rearranged light chains were found to have identical amino acids. These studies suggest that there is no absolute restriction in the use of V region germline genes and form the foundation for understanding the humoral immune response to the CD4-binding site of gp120.

Relative Recovery of Thermal Energy and Fresh Water in Aquifer Storage and Recovery Systems.

This paper explores the relationship between thermal energy and fresh water recoveries from an aquifer storage recovery (ASR) well in a brackish confined aquifer. It reveals the spatial and temporal distributions of temperature and conservative solutes between injected and recovered water. The evaluation is based on a review of processes affecting heat and solute transport in a homogeneous aquifer. In this simplified analysis, it is assumed that the aquifer is sufficiently anisotropic to inhibit density-affected flow, flow is axisymmetric, and the analysis is limited to a single ASR cycle. Results show that the radial extent of fresh water at the end of injection is greater than that of the temperature change due to the heating or cooling of the geological matrix as well as the interstitial water. While solutes progress only marginally into low permeability aquitards by diffusion, conduction of heat into aquitards above and below is more substantial. Consequently, the heat recovery is less than the solute recovery when the volume of the recovered water is lower than the injection volume. When the full volume of injected water is recovered the temperature mixing ratio divided by the solute mixing ratio for recovered water ranges from 0.95 to 0.6 for ratios of maximum plume radius to aquifer thickness of 0.6 to 4.6. This work is intended to assist conceptual design for dual use of ASR for conjunctive storage of water and thermal energy to maximize the potential benefits.

Use of polymerase chain reaction for the rapid construction of synthetic genes.

Although the polymerase chain reaction (PCR) (1,2) is invaluable for the cloning and manipulation of existing DNA sequences, PCR also makes it possible to create new DNA fragments consisting of a nucleic acid sequence that is specified entirely by the investigator. In this chapter we describe a simple two-step PCR method for the rapid construction of synthetic genes (3). This method is based on early observations by Mullis et al. (4) in which multiple overlapping oligonucleotides could be used to generate synthetic DNA through several sequential rounds of Klenow-based PCR amplification. The method described in this chapter utilizes the thermostabile Taq polymerase and allows for the generation of synthetic genes in as little as 1 d. This method has proven useful in studies in which synthetic genes were constructed for the HIV-2 Rev protein (3,5) and the Wilms' tumor locus zinc finger protein (6). Furthermore, this method has been successfully employed in extensive mutagenesis of the HIV-1 rev response element (7).

Response of the Plastic Lake catchment, Ontario, to reduced sulphur deposition.

As a consequence of decreases in the emission rate of sulphur in eastern North America in the late 1970s and early 1980s, sulphate deposition in central Ontario declined by about 40%, but has remained constant for about six years. Plastic Lake, a small, dilute lake on the Precambrian shield that the authors have studied since 1979, acidified between the start of the study and about 1986, but since then has not changed. The authors also monitored the chemistry of streamwater draining the Plastic Lake catchment. Water quality of runoff from an upland site improved rapidly (pH and alkalinity increased, SO4(2-) and Al decreased), but two factors offset these improvements. A small wetland area downstream reversed most of these changes, resulting in a constant output of strong acid from the catchment. In addition, in extremely dry years (1983, 1987, 1989) there were very high concentrations of SO4(2-) in the streamwater, suggesting substantial re-oxidation of reduced S in the catchment.

Comparison of iron accumulation in lakes using sediment core and mass balance calculations.

The accumulation of iron (Fe) in several lakes in Ontario, Canada was determined by two independent approaches. First, Fe accumulation was calculated in cores collected from several sites in each lake by integrating Fe concentration profiles with sediment accumulation rates determined from Pb210 dating. These site-specific accumulation rates were corrected for sediment focussing so that whole-lake Fe accumulation values could be derived. Using this approach, recent whole-lake Fe accumulation in eight lakes ranged between approximately 750 and 4000 mg/m2 per year. Second, whole-lake Fe accumulation was estimated from lake mass budgets, which were measured over a maximum of 14 years. Accumulation measured using the mass balances ranged from 10 to 1330 mg/m2 per year. Comparison of the two approaches indicated that retentions calculated from the sediment cores were much greater than those estimated from the mass balances. The most likely explanation for this difference is that, in the two decades since the cores were collected, there has been a substantial decline in Fe retention (in mass units but not percent) in the study lakes, principally as a result of reduced inputs of Fe from the catchments.

Uncertainty assessments and hydrological implications of climate change in two adjacent agricultural catchments of a rapidly urbanizing watershed.

Lake Simcoe is the most important inland lake in Southern Ontario. The watershed is predominantly agricultural and under increasing pressure from urbanization, leading to changing runoff patterns in rivers draining to the lake. Uncertainties in rainfall-runoff modeling in tributary catchments of the Lake Simcoe Watershed (LSW) can be an order of magnitude larger than pristine watersheds, hampering water quality predictions and export calculations. Here we conduct a robust assessment to constrain the uncertainty in hydrological simulations and projections in the LSW using two representative adjacent agricultural catchments. Downscaled CGCM 3 projections using A1B and A2 emission scenarios projected increases of 4°C in air temperature and a 26% longer growing season. The fraction of precipitation falling as snow will decrease. Spring runoff is an important event in LSW but individual HBV best calibrated parameter sets under-predicted peak flows by up to 32%. Using an ensemble of behavioral parameter sets achieved credible representations of present day hydrology and constrained uncertainties in future projections. Parameter uncertainty analysis showed that the catchments differ in terms of their snow accumulation/melt and groundwater dynamics. Human activities exacerbate the differences in hydrological response. Model parameterization in one catchment could not generate credible hydrological simulations in the other. We cautioned against extrapolating results from monitored to ungauged catchments in managed watersheds like the LSW.

Modelling the long term impact of climate change on the carbon budget of Lake Simcoe, Ontario using INCA-C.

This study presents a process-based model of dissolved organic carbon concentration ([DOC]) in catchments draining into Lake Simcoe, Ontario. INCA-C, the Integrated Catchment model for Carbon, incorporates carbon biogeochemical processes in a terrestrial system with hydrologic flow paths to simulate watershed wide [DOC]. The model successfully simulates present-day inter-annual and seasonal [DOC] dynamics in tributaries draining catchments with mixed or contrasting land cover in the Lake Simcoe watershed (LSW). The sensitivity of INCA-C to soil moisture, hydrologic controls and land uses within a watershed demonstrates its significance as a tool to explore pertinent environmental issues specific to the LSW. Projections of climate change under A1B and A2 SRES scenarios suggest a continuous monotonic increase in [DOC] in surface waters draining into Lake Simcoe. Large variations in seasonal DOC dynamics are predicted to occur during summer with a possibility of displacement of summer [DOC] maxima towards winter and a prolongation of summer [DOC] levels into the autumn. INCA-C also predicts possible increases in dissolved inorganic carbon in some tributaries with rising temperature suggesting increased CO(2) emissions from rivers as climate changes.

Using the INCA-Hg model of mercury cycling to simulate total and methyl mercury concentrations in forest streams and catchments.

We present a new, catchment-scale, process-based dynamic model for simulating mercury (Hg) in soils and surface waters. The Integrated Catchments Model for Mercury (INCA-Hg) simulates transport of gaseous, dissolved and solid Hg and transformations between elemental (Hg(0)), ionic (Hg(II)) and methyl (MeHg) Hg in natural and semi-natural landscapes. The mathematical description represents the model as a series of linked, first-order differential equations describing chemical and hydrological processes in catchment soils and waters which we believe control surface water Hg dynamics. The model simulates daily time series between one and 100 years long and can be applied to catchments ranging in size from <1 to ~10,000 km(2). Here we present applications of the model to two boreal forest headwater catchments in central Canada where we were able to reproduce observed patterns of stream water total mercury (THg) and MeHg fluxes and concentrations. Model performance was assessed using Monte Carlo techniques. Simulated in-stream THg and MeHg concentrations were sensitive to hydrologic controls and terrestrial and aquatic process rates.