A bioassay to assess the potential effects of sediment resuspension on phytoplankton community composition

A laboratory assay (SAGA or Sediment Algal Growth Assay) was developed to assess the potential impact of sediment resuspension on the structure of phytoplankton communities, and to evaluate the effectiveness of various sediment treatments in decreasing the abundance of blue-green algae in the event of sediment resuspension during storms. In assays with sediment from eutrophic Akanoi Bay, Lake Biwa, Japan, 7–11 species of phytoplankton seeded from the sediments grew during the 3-week assay indicating that sediment resuspension has the potential to increase both phytoplankton biomass and species diversity. Treatment of sediments with Ca(NO3)2 substantially decreased phytoplankton biomass (measured as chlorophyll concentration) in assays with sediments from Akanoi Bay and the North Basin of Lake Biwa. Further, among various oxidation treatments of sediments, Ca(NO3)2 was most effective in decreasing or preventing filamentous blue-green algal growth in N- and P-replete media. In contrast, when sediments were added to P-limited phytoplankton dominated by green algae and diatoms, no growth of blue-green algae occurred regardless of sediment treatment. This revised version was published online in June 2006 with corrections to the Cover Date.     

The effect of heavy metal speciation in sediment on bioavailability to tubificid worms

The bioavailability of heavy metals in sediment to freshwater tubificid worms was compared with measures of chemical extractability using a sequential extraction procedure. In order to provide a range of test sediments of different quality, various mineral phases were prepared, in which the metals were spiked by adsorption or coprecipitation and these were then mixed with a bulk base sediment in known proportions. Results indicated good correlation between worm metal burden and metal mobilised from the sediments in the first (exchangeable) sequential extraction step for Cd, Cu and Pb. Of the other metals tested, Zn levels in the worms were found to be constant, suggesting regulation, and Ni uptake was too small for accurate measurement. In general, metals spiked to the sediment directly, or adsorbed on the clay mineral phase were found to be much more available than those bound to sewage sludge, carbonate or hydrous ferric oxide phases.     

Tubifex tubifex as a link in food chain transfer of hexachlorobenzene from contaminated sediment to fish

Sediments contaminated with poorly water-soluble organic chemicals pose a risk to aquatic food chains. Sediment-associated chemicals can be accumulated by endobenthic, sediment-ingesting invertebrates. Some tubificid species – or other benthic annelids – serve as food for benthivorous fish, which thereby ingest the sediment-borne chemicals and may accumulate contaminant concentrations far higher than from water exposure only, and transfer them to organisms of higher trophic levels. For measurement of biomagnification, a sediment based food chain was developed and established in the laboratory. The two-step food chain included the sediment-dwelling freshwater oligochaete Tubifex tubifex (Müller) as a representative species of benthic infauna. The three-spined stickleback (Gasterosteus aculeatus, Linné), a small teleost fish which often feeds primarily on benthic invertebrates, served as a model predator. Spiked artificial sediment and reconstituted water as the overlying medium were used. Experiments were performed using ¹⁴C-labelled hexachlorobenzene, a hydrophobic pollutant as a model compound. To examine the influence of benthic prey on the bioaccumulation of the test substance in the predator, fish were exposed to spiked water, spiked sediment, pre-contaminated prey organisms, or combinations of these exposure routes. The results of these experiments indicate that for hexachlorobenzene, the presence of contaminated Tubifex tubifex as a food source in combined exposure leads to significantly higher accumulation in fish than exposure to single pathways.     

Development of a biotic index using stream macroinvertebrates to assess stress from deposited fine sediment

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Reduced territorial defence in rainbow trout fry exposed to a paper mill effluent: using the mirror image stimulation test as a behavioural bioassay

Rainbow trout Oncorhynchus mykiss fry exposed to a paper mill effluent showed increased EROD (ethoxyresorufin-O-deethylase) activity and reduced territorial aggression in mirror image stimulation (MIS) tests. These results suggest that the MIS test could be a useful tool to detect sublethal behavioural effects of pollution in fish fry.     

Distribution and location of polycyclic aromatic hydrocarbons (PAHs) and PAH-degrading bacteria within polluted soil aggregates

A study was conducted to determine the location and distribution of PAH and PAH-degrading bacteria in different aggregate size fractions of an industrially polluted soil. The estimation of PAH-degrading bacteria using an MPN microplate technique indicated that these bacteria are most numerous in the aggregate size fractions corresponding to fine silt (2–20m) and clay(<2m) compared to larger fractions or unfractionated soil.PAH concentrations were also highest in the aggregate size fraction corresponding to fine silt. Similar results were found in a spiked soil (incubated for 6 months) with similar carbonated minerals. Transmission electron microscopy observations showed that the autochtonous PAH-degrading bacteria were embedded in the aggregates where PAHs were abundant. In spite of this extensive co-localisation PAH degradation was limited during 6 months incubation. This indicates that factors other than spatial distribution and PAH degrading ability control degradation rates. The fine silt fraction of the industrial soil had an elevated C/N ratio (35) compared to the clay fraction (C/N: 16). Thus the fraction which assumably had the highest specific surface area contained less PAH but similar numbers of PAH-degraders. N thus seem to play an important role in the long term, but as PAH degradation was low in fine size fractions, other sources/factors were probably limiting (easily degradable C, P org, O₂ etc.). Based on these findings, soil particle organization and structure of soil aggregates appear to be important for the characterization of a polluted soil (localization and sequestration). Manipulations that modify aggregation in polluted soils could thus potentially influence the accessibility and biodegradability of PAHs.     

A new standardized sediment bioassay protocol using the amphipod Hyalella azteca (Saussure)

A new standardized bioassay procedure for testing the chronic toxicity of sediments to Hyalella azteca was developed. Tests were initiated with 0–1 wk old amphipods exposed to sediments from Hamilton Harbour, Toronto Harbour, and Lake Ontario for 4 to 8 weeks. Both survival and growth were significantly reduced in the Hamilton Harbour sediments relative to those from the lake after 4 weeks. Exposures of 8 weeks resulted in greater variability; survival of amphipods in sediments from one of the harbour stations, and growth in sediments from both harbour stations with surviving young were not statistically different from survival and growth in lake sediments. Growth and survival in lake sediments were comparable to cultures grown with cotton gauze and no sediment after 4 weeks, but survival was poorer by week 8. Replication was good in 12 out of 13 tests done in duplicate; the difference in survival between replicates averaged 2.2 animals (20 amphipods/replicate, 4 week exposure). We propose that 4 week exposures of young (0–1 wk old) Hyalella would provide a suitable standardized chronic toxicity test for sediments. A detailed protocol on the methodology is presented.     

DNA extraction for 16S rRNA gene analysis to determine genetic diversity in deep sediment communities

A protocol was devised which permitted the extraction of DNA from deep marine sediments up to 503 m below the sea floor. These sediments have been laid down over the last 3 million years. 16S rRNA gene sequences were amplified from the DNA by the polymerase chain reaction. The details of the successful extraction and polymerase chain reaction methodology varied between samples from different depths. This emphasizes the attention to detail required to allow the diversity of bacteria in these deep sediments to be studied.     

Mouse embryos used as a bioassay to determine control of marsupial embryonic diapause

Mouse blastocysts appear to be under direct inhibition from the uterine environment, whereas no evidence of direct inhibition during diapause in the tammar wallaby has been observed. Normally developing (day 4) and quiescent mouse blastocysts were incubated for up to 12 hr in media supplemented with BSA, wallaby plasma, wallaby day 0 (day of removal of pouch young; RPY), day 5, or day 10 endometrial exudates at a concentration of 2 mg/ml of protein, and analyzed for rates of carbohydrate metabolism using fluorescence and radioisotopes. Rates of glucose uptake and lactate production by day 4 blastocysts increase after incubation with day 10 and day 5 wallaby exudates compared with rates by blastocysts incubated in BSA. Pyruvate uptake increased after 8 hr irrespective of incubation media, except for embryos incubated in day 0 exudate, which maintained levels significantly lower than BSA-incubated embryos. Quiescent mouse embryos displayed a high ATP/ADP ratio during diapause (1.06 +/- 0.24) which decreased after 4 hr incubation in all media (0.42 +/- 0.05; P < 0.01) but embryos incubated in day 0 exudate media remained at a significantly higher level than embryos incubated in BSA. These results indicate that quiescent tammar endometrial exudate is not capable of initiating diapause in mouse embryos at the concentration used, but is able to slow the rate of reactivation of quiescent blastocysts. Importantly, reactivated wallaby exudate increases mouse blastocyst glucose metabolism and lactate production. It is possible that the quiescent tammar endometrial environment has an inhibitory factor necessary to maintain diapause in the tammar blastocyst.     

Effects of sediment loads on the fish and invertebrates of a Sierra Nevada river, California

This study examined the effects of sedimentloadings from an abandoned hydraulic miningsite on potential trout spawning gravels, largeaquatic macroinvertebrates, and fish growth,survival and reproduction in the South YubaRiver (Nevada County) California. Effects ofsediment loadings, which are transported to theSouth Yuba River via Humbug Creek, wereinvestigated by comparing data from sites onthe South Yuba River upstream and downstream ofHumbug Creek. The study did not find anydeleterious effects of sediment loadings onfish survival and reproduction (based onsnorkel survey data), or large aquaticmacroinvertebrate populations. In addition,the lack of a significant difference in thepercentage of 0.30 to 3.35 mm material insubstrate samples suggests that sediment loadsare not affecting trout spawning gravels. Incontrast, Sacramento pikeminnow(Ptychocheilus grandis) growth appears to beaffected by both water temperatures andsediment loadings. Downstream of Humbug Creek,growth during the first year was significantlyfaster, based on calculated standard lengths atthe age 1 annulus, but condition factor wassignificantly lower than above Humbug Creek.     

Use of microcosms to determine persistence of Escherichia coli in recreational coastal water and sediment and validation with in situ measurements

AIMS: To determine the persistence of the faecal indicator organism Escherichia coli in recreational coastal water and sediment using laboratory-based microcosms and validation with in situ measurements. METHODS AND RESULTS: Intact sediment cores were taken from three distinct coastal sites. Overlying estuarine water was inoculated with known concentrations of E. coli and decay rates from both overlying water and sediment were determined following enumeration by the membrane filtration method at fixed time intervals over a 28-day period. It was demonstrated that E. coli may persist in coastal sediment for >28 days when incubated at 10 degrees C. Escherichia coli survival was found to have an inverse relationship with temperature in both water and sediment. In general the decay rate for E. coli was greater in water than in sediment. Small particle size and high organic carbon content were found to enhance E. coli survival in coastal sediments in the microcosms. CONCLUSIONS: Results of this microcosm study demonstrated the more prolonged survival of E. coli in coastal sediments compared with overlying water, which may imply an increased risk of exposure because of the possible resuspension of pathogenic micro-organisms during natural turbulence or human recreational activity. SIGNIFICANCE AND IMPACT OF THE STUDY: A more accurate estimate of exposure risk has been described which may subsequently be used in a quantitative microbial risk assessment for recreational coastal waters.     

Use of a multiple addition bioassay to determine limiting nutrients in eagle lake,California

This investigation uses anin vitro enrichment bioassay (in which all essential nutrients except one are added to each culture) to determine which nutrients are important to algal growth in Eagle Lake. The technique was developed when it was discovered that addition of individual nutrients produced little if any growth response. Laboratory bioassays correlated well with comparative studies in the lake. A great deal of variation was found throughout the year but P, N, Fe, and S were found to be limiting at one time or another. The north and south basins of the lake, which differ in morphometry, were also found to differ in the intensity spectrum of limitation. While P was the most important nutrient in both basins, the other nutrients were more limiting in the north basin than the south, and Fe, which was least limiting in the south, was very important in the north. The multiple enrichment bioassay has several advantages over other bioassays. Supported in part by Research Corporation     

Substrate roughness, fish grazers, and mesohabitat type interact to determine algal biomass and sediment accrual in a high-altitude subtropical stream

Since periphytic biofilm is an important source of food in lotic ecosystems, it is important to understand how key ecological factors affect the accrual and loss of algal biomass and sediment in the biofilm. We designed a field experiment to evaluate the effects of mesohabitat type (pools and riffles), grazing fish (control and exclusion), and substrate roughness (smooth and rough) on chlorophyll a, ash-free dry mass (AFDM), and total dry mass in a subtropical stream. Mesohabitat type did not influence the effect of grazers on periphyton. However, rough substrates accumulated more total dry mass in pools than in riffles, while smooth substrates accumulated similar amounts of total dry mass in both mesohabitats. The accrual of AFDM and chlorophyll a was greater on rough than on smooth substrates, regardless of mesohabitat. Treatments without fish accrued more total dry mass, AFDM, and chlorophyll a than treatments with fish, showing that fish play a major role in this stream by removing sediment and algal biomass. These results suggest that habitat simplification in the scale of substrate roughness and loss of large grazers may impact the accrual and loss of algal biomass and sediment in lotic ecosystems.     

Development and assessment of indices to determine stream fish vulnerability to climate change and habitat alteration

Understanding the future impacts of climate and land use change are critical for long-term biodiversity conservation. We developed and compared two indices to assess the vulnerability of stream fish in Missouri, USA based on species environmental tolerances, rarity, range size, dispersal ability and on the average connectivity of the streams occupied by each species. These two indices differed in how environmental tolerance was classified (i.e., vulnerability to habitat alteration, changes in stream temperature, and changes to flow regimes). Environmental tolerance was classified based on measured species responses to habitat alteration, and extremes in stream temperatures and flow conditions for one index, while environmental tolerance for the second index was based on species’ traits. The indices were compared to determine if vulnerability scores differed by index or state listing status. We also evaluated the spatial distribution of species classified as vulnerable to habitat alteration, changes in stream temperature, and change in flow regimes. Vulnerability scores were calculated for all 133 species with the trait association index, while only 101 species were evaluated using the species response index, because 32 species lacked data to analyze for a response. Scores from the trait association index were greater than the species response index. This is likely due to the species response index's inability to evaluate many rare species, which generally had high vulnerability scores for the trait association index. The indices were consistent in classifying vulnerability to habitat alteration, but varied in their classification of vulnerability due to increases in stream temperature and alterations to flow regimes, likely because extremes in current climate may not fully capture future conditions and their influence on stream fish communities. Both indices showed higher mean vulnerability scores for listed species than unlisted species, which provided a coarse measure of validation. Our indices classified species identified as being in need of conservation by the state of Missouri as highly vulnerable. The distribution of vulnerable species in Missouri showed consistent patterns between indices, with the more forest-dominated, groundwater fed streams in the Ozark subregion generally having higher numbers and proportions of vulnerable species per site than subregions that were agriculturally dominated with more overland flow. These results suggest that both indices will identify similar habitats as conservation action targets despite discrepancies in the classification of vulnerable species. Our vulnerability assessment provides a framework that can be refined and used in other regions.     

Search of Microorganisms that Degrade PAHs under Alkaline Conditions

Bacterial strains were enriched from building rubble contaminated with polycyclic aromatic hydrocarbons (PAHs). These strains were studied as an inoculum in bioremediation processes with contaminated building rubble. The selection criteria for the bacteria were broad profiles in PAH degradation, stable expression of the traits and tolerance to alkaline conditions. Various strains of Micrococcus sp., Dietzia sp., Rhodococcus sp. and Pseudomonas sp. met the selection criteria. In general, degradative activity was limited at higher pH values. Strains of Micrococcus were suitable for practical use as complete degradation of various PAHs was observed at pH values exceeding 10. Strains of Dietzia sp. showed broad PAH degradation profile, but in some cases degradation came to a halt leaving some of the PAHs unutilized. With Dietzia sp. this could be due to inhibitory effects from the accumulation of toxic PAH metabolic products and/or growth‐limiting media conditions.     

Seasonality of sulfate reduction and pore water solutes in a marine fish farm sediment: the importance of temperature and sedimentary organic matter

Sulfate reduction and pore water solutes related to sulfur cycling and anaerobic processes (short chain fatty acids (SCFA), SO₄ ^2–, TCO₂, NH₄ ⁺, dissolved sulfides (H₂S) and CH₄) were examined during one year at a marine fish farm. Mineralization of fish farm waste products was rapid in this non-bioturbated, organic rich sediment. Stimulation of sulfate reduction rates (SRR) occurred primarily in the surface layers where the organic matter was deposited. Acetate was the most important (<99%) of the measured SCFA attaining high concentrations during summer months (up to 4.7 mM). The acetate profiles exhibited distinct seasonal cycles, where periods with high concentrations in the pore waters were found coincident with a high pool of particulate organic matter in the surface sediments and a low activity of the sulfate reducing bacteria (early spring and late summer). Periods with low acetate pools occurred when sulfate reduction rates were high in early summer and in winter were pools of particulate organic matter were decreasing. Methane production was observed concurrent with sulfate reduction in the microbial active surface layers in late summer. Subsurface peaks of SO₄ ^2–, TCO₂, NH₄ ⁺ and H₂S were evident in July and August due to rapid mineralization in these surface layers. With decreasing autumn water temperatures mineralization rates declined and subsurface peaks of these solutes disappeared. A strong relationship was found between pore water TCO₂, and NH₄ ⁺. Ratios between TCO₂, and NH₄ ⁺ were low compared to a control site, attaining minimum values in mid-summer. This indicated rapid nitrogen mineralization of nitrogen rich labile substrates in the fish farm sediment during the entire season.     

Fluoranthene as a model toxicant in sediment studies with Chironomus riparius

The bioavailability of fluoranthene (as a model toxicant) in freshwater sediment tests with Chironomus riparius was investigated in relation to the partitioning of the chemical between water and sediment. Effects on survival and successful development of newly hatched larvae through to adult emergence were measured over a range of sediment concentrations. Radiolabelled fluoranthene was employed to enable sediment, aqueous and tissue concentrations to be determined. A significant effect on emergence was obtained at a measured sediment concentration of 80 mg kg^-1 (dry weight). The measured concentration in the overlying water was 88 g l^-1. Although effects at 170 mg kg^-1 (dry weight) were more severe, the measured concentration in the overlying water was similar (89 g l^-1), probably reflecting the approach towards the solubility limit. Bioaccumulation of fluoranthene, determined by analysis of emerging adults, was approximately proportional to the sediment concentration. Although the aqueous phase concentration appeared to be the principal factor controlling bioavailability, the increasing toxicity, when water concentrations had reached a plateau, suggested that other mechanisms may also be involved. Different sediment spiking procedures resulted in broadly similar partitioning of the chemical between the sediment and the overlying water. There were only minor differences in toxicity between the procedures, two of which allowed the carrier solvent used for spiking to be eliminated before addition of the test organisms. It was concluded that, for most purposes, all four procedures were equally acceptable, and by retaining a flexible approach, even for standardised regulatory tests, spiking procedures may be tailored to the physico-chemical properties of the test material.     

A study to determine the impact of IMPT optimization techniques on prostate synthetic CT image sets dose comparison against CT image sets

BackgroundThe objective of this study is to determine the impact of intensity modulated proton therapty (IMPT) optimization techniques on the proton dose comparison of commercially available magnetic resonance for calculating attenuation (MRCA T) images, a synthetic computed tomography CT (sCT) based on magnetic resonance imaging (MRI) scan against the CT images and find out the optimization technique which creates plans with the least dose differences against the regular CT image sets.Material and methodsRegular CT data sets and sCT image sets were obtained for 10 prostate patients for the study. Six plans were created using six distinct IMPT optimization techniques including multi-field optimization (MFO), single field uniform dose (SFUD) optimization, and robust optimization (RO) in CT image sets. These plans were copied to MRCA T, sCT datasets and doses were computed. Doses from CT and MRCA T data sets were compared for each patient using 2D dose distribution display, dose volume histograms (DVH), homogeneity index (HI), conformation number (CN) and 3D gamma analysis. A two tailed t-test was conducted on HI and CN with 5% significance level with a null hypothesis for CT and sCT image sets.ResultsAnalysis of ten CT and sCT image sets with different IMPT optimization techniques shows that a few of the techniques show significant differences between plans for a few evaluation parameters. Isodose lines, DVH, HI, CN and t-test analysis shows that robust optimizations with 2% range error incorporated results in plans, when re-computed in sCT image sets results in the least dose differences against CT plans compared to other optimization techniques. The second best optimization technique with the least dose differences was robust optimization with 5% range error.ConclusionThis study affirmatively demonstrates the impact of IMPT optimization techniques on synthetic CT image sets dose comparison against CT images and determines the robust optimization with 2% range error as the optimization technique which gives the least dose difference when compared to CT plans.     

Short review of selected fish biomarkers of xenobiotic exposure with an example using fish hepatic mixed-function oxidase

Abstract One group of biological tools that are useful for monitoring exposure to xenobiotics (and hence water quality) have been collectively referred to as biomarkers and are denned in this paper as any biochemical, histological and/or physiological alterations or manifestations of stress. Biomarkers within an aquatic toxicological context generally represent biological responses of individual organisms to xenobiotic exposure (i. e. responses at the whole organism level of biological organization). These include among others, enzyme alterations, bile metabolites, RNA/DNA ratio, adenylate energy charge, skeletal abnormalities, immune dysfunction, behavioural changes and histopathological lesions. Biomarkers can act as effective early warning sentinels to ensure the protection of the integrity of whole ecosystems, including freshwater and marine ecosystems. This paper briefly reviews a selection of fish biomarkers of xenobiotic chemical exposure and discusses their respective strengths and limitations for use in biomonitoring. An example of the application of fish mixed-function oxidase (MFO) and cytochrome P-450 as biomarkers of chemical exposure in Port Phillip Bay is provided. It is concluded that judicious application of biomarkers such as MFO in association with an understanding of the underlying causal mechanisms of induction and toxicity, will contribute to the successful prediction of biological effects of xenobiotic exposure on fish population health.     

Spheroidal carbonaceous particles as a marker for recent sediment distribution

We have tested the hypothesis that spheroidal carbonaceous particles (SCP), emitted from oil and coal combustion, can be used as a marker for recent sediment distribution in lake basins. Sediment traps have been used to study when and how the particles are introduced into a lake, a laboratory experiment has been made to study the depositional behaviour of the particles in relation to other sedimentary constituents, and spatial SCP distributions have been surveyed in three lakes. The results support the hypothesis that SCP can be used as a marker, and also indicate that acidification can change sediment distribution in lakes.