The influence of time on lead toxicity and bioaccumulation determined by the OECD earthworm toxicity test

Internationally agreed standard protocols for assessing chemical toxicity of contaminants in soil to worms assume that the test soil does not need to equilibrate with the chemical to be tested prior to the addition of the test organisms and that the chemical will exert any toxic effect upon the test organism within 28 days. Three experiments were carried out to investigate these assumptions. The first experiment was a standard toxicity test where lead nitrate was added to a soil in solution to give a range of concentrations. The mortality of the worms and the concentration of lead in the survivors were determined. The LC(50)s for 14 and 28 days were 5311 and 5395 mug(Pb) g(soil)(-1) respectively. The second experiment was a timed lead accumulation study with worms cultivated in soil containing either 3000 or 5000 mug(Pb) g(soil)(-1). The concentration of lead in the worms was determined at various sampling times. Uptake at so' Sol both concentrations was linear with time. Worms in the 5000 mug g(-1) soil accumulated lead at a faster rate (3.16 mug Pb g(tissue)(-1) day(-1)) tiss than those in the 3000 mug g(-1) soil (2.21 mug Pb-tissue g(-1) day(-1)). The third experiment was a timed experiment with worms cultivated in tiss soil containing 7000 mugPb g(soil)(-1). Soil and lead nitrate solution were mixed and stored at 20 degreesC. Worms were added at various times over a 35-day period. The time to death increased from 23 h, when worms were added directly after the lead was added to the soil, to 67 It when worms were added after the soil had equilibrated with the lead for 35 days. In artificially Pb-amended soils the worms accumulate Pb over the duration of their exposure to the Pb. Thus time limited toxicity tests may be terminated before worm body load has reached a toxic level. This could result in under-estimates of the toxicity of Pb to worms. As the equilibration time of artificially amended Pb-bearing soils increases the bioavailability of Pb decreases. Thus addition of worms shortly after addition of Pb to soils may result in the over-estimate of Pb toxicity to worms. The current OECD acute worm toxicity test fails to take these two phenomena into account thereby reducing the environmental relevance of the contaminant toxicities it is used to calculate. (C) 2002 Elsevier Science Ltd. All rights reserved.

A refined LEED analysis of water on Ru{0001}: an experimental test of the partial dissociation model

Despite a number of earlier studies which seemed to confirm molecular adsorption of water on close-packed surfaces of late transition metals, new controversy has arisen over a recent theoretical work by Feibelman, according to which partial dissociation occurs on the Ru{0001} surface leading to a mixed (H2O + OH + H) superstructure. Here, we present a refined LEED-IV analysis of the (root3 x root3)R30degrees-D2O-Ru{0001} structure, testing explicitly this new model by Feibelman. Our results favour the model proposed earlier by Held and Menzel assuming intact water molecules with almost coplanar oxygen atoms and out-of-plane hydrogen atoms atop the slightly higher oxygen atoms. The partially dissociated model with an almost identical arrangement of oxygen atoms can, however, not unambiguously be excluded, especially when the single hydrogen atoms are not present in the surface unit cell. In contrast to the earlier LEED-IV analysis, we can, however, clearly exclude a buckled geometry of oxygen atoms.

The effect of (mis-specified) GARCH filters on the finite sample distribution of the BDS test

This paper considers the effect of using a GARCH filter on the properties of the BDS test statistic as well as a number of other issues relating to the application of the test. It is found that, for certain values of the user-adjustable parameters, the finite sample distribution of the test is far-removed from asymptotic normality. In particular, when data generated from some completely different model class are filtered through a GARCH model, the frequency of rejection of iid falls, often substantially. The implication of this result is that it might be inappropriate to use non-rejection of iid of the standardised residuals of a GARCH model as evidence that the GARCH model ‘fits’ the data.

Nitrogen fertilizer and seed rate effects on Hagberg failing number of hybrid wheats and their parents are associated with alpha-amylase activity, grain cavity size and dormancy

Field experiments were carried out to assess the effects of nitrogen fertilization and seed rate on the Hagberg falling number (HFN) of commercial wheat hybrids and their parents. Applying nitrogen (200 kg N ha(-1)) increased HFN in two successive years. The HFN of the hybrid Hyno Esta was lower than either of its parents (Estica and Audace), particularly when nitrogen was not applied. Treatment effects on HFN were negatively associated with a-amylase activity. Phadebas grain blotting suggested two populations of grains with different types of a-amylase activity: Estica appeared to have a high proportion of grains with low levels of late maturity endosperm a-amylase activity (LMEA); Audace had a few grains showing high levels of germination amylase; and the hybrid, Hyno Esta, combined the sources from both parents to show heterosis for a-amylase activity. Applying nitrogen reduced both apparent LMEA and germination amylase. The effects on LMEA were associated with the size and disruption of the grain cavity, which was greater in Hyno Esta and Estica and in zero-nitrogen treatments. External grain morphology failed to explain much of the variation in LMEA and cavity size, but there was a close negative correlation between cavity size and protein content. Applying nitrogen increased post-harvest dormancy of the grain. Dormancy was greatest in Estica and least in Audace. It is proposed that effects of seed rate, genotype and nitrogen fertilizer on HFN are mediated through factors affecting the size and disruption of the grain cavity and therefore LMEA, and through factors affecting dormancy and therefore germination amylase. (c) 2004 Society of Chemical Industry.

Influence of temperature on the production and infectivity of entomopathogenic nematodes against larvae and pupae of vine weevil, Otiorhynchus sulcatus, (Coleoptera: Curculionidae)

Susceptibility of late instar vine weevil Otiorhynchus sulcatus larvae and pupae to four species entomopathogenic nematodes were tested. Bioassays on production and infectivity to larvae and pupae were compared for two steinernematids and two heterorhabditis such as Steinernema carpocapsae, S. feltiae, Heterorhabditis indica and H. bacteriophora. Nematodes production of all species was determined by the number infective juveniles (IJs) established in vine weevil larvae and pupae O. sulcatus using sand and filter paper bioassay. S. feltiae produced the maximum number in larvae and pupae at 20°C as compared to other nematodes but production of H. indica, was better at 25°C in larvae and pupae followed by H. bacteriophora, S. carpocapsae and Infectivity test of larvae and pupae was also done in sand media. Infective juveniles recovered from larvae and pupae when infected with S. feltiae produced maximum infective juveniles at 20°C temperatures than all other isolates. H. bacteriophora produced higher number of IJs in larvae and pupae than all other nematode isolates at 25°C. This paper indicates the application of nematodes with the knowledge of insect pest biology represents a possible new strategy for O. sulcatus larvae and pupae.

Some factors affecting reproductive success in crossbred dairy cows on smallholder farms in coastal north-east Tanzania

A 2-year longitudinal survey was carried out to investigate factors affecting reproduction in crossbred cows on smallholder farms in and around an urban centre. Sixty farms were visited at approximately 2-week intervals and details of reproductive traits and body condition score (BCS) were collected. Fifteen farms were within the town (U), 23 farms were approximately 5 km from town (SU), and 22 farms approximately 10 km from town (PU). Sources of variation in reproductive traits were investigated using a general linear model (GLM) by a stepwise forward selection and backward elimination approach to judge important independent variables. Factors considered for the first step of formulation of the model included location (PU, SU and U), type of insemination, calving season, BCS at calving, at 3 months postpartum and at 6 months postpartum, calving year, herd size category, source of labour (hired and family labour), calf rearing method (bucket and partial suckling) and parity number of the cow. The effects of the independent variables identified were then investigated using a non-parametric survival technique. The number of days to first oestrus was increased on the U site (p = 0.045) and when family labour was used (p = 0.02). The non-parametric test confirmed the effect of site (p = 0.059), but effect of labour was not significant. The number of days from calving to conception was reduced by hiring labour (p = 0.003) and using natural service (p = 0.028). The non-parametric test confirmed the effects of type of insemination (p = 0.0001) while also identifying extended calving intervals on U and SU sites (p = 0.014). Labour source was again non-significant. Calving interval was prolonged on U and SU sites (p = 0.021), by the use of AI (p = 0.031) and by the use of family labour (p = 0.001). The non-parametric test confirmed the effect of site (p = 0.008) and insemination type (p > 0.0001) but not of labour source. It was concluded that under favourable conditions (PU site, hired labour and natural service) calving intervals of around 440 days could be achieved.

Histone H4 gene expression and morphological changes on shoot apices of strawberry (Fragaria x ananassa Duch.) during floral induction

To investigate flower induction in June-bearing strawberry plants, morphological changes in shoot apices and Historic H4 expression in the central zone during flower initiation were observed. Strawberry plants were placed under flower inducible, short-day conditions (23 degrees C/17 degrees C, 10 h day length) for differing number of days (8, 16, 20, 24 or 32 days) and then these plants were transferred to non-inducible, long-day conditions (25 degrees C/20 degrees C, 14 h day length). The shoot apices of plants placed under short-day conditions for 8 days were flat, similar to shoot apices of plants in the vegetative phase of development, and Histone H4 was not expressed in the central zone during the experimental period. On the other hand, the shoot apices of plants placed under short-day conditions for 16 days remained flat, similar to shoot apices of plants placed under short-day conditions for 8 days, but Histone H4 was expressed in the central zone at the end of the short-day treatment. Morphological changes in the shoot apices of these plants were observed 8 days after the change in day-length. These plants developed differentiated flower organs after they were grown for another 30 days under long-day conditions. These results indicate that changes in the expression pattern of the Histone H4 gene occur before morphological changes during flower induction and that the expression of the gene in the central zone can be used as one of the indicators of the flowering process in strawberries. (c) 2006 Elsevier B.V. All rights reserved.

In vitro examination of the effect of Orlistat on the ability of the faecal microbiota to utilize dietary lipids

Orlistat is an anti-obesity treatment with which several gastrointestinal (GI) side-effects are commonly associated in the initial stages of therapy. There is no physiological explanation as to why two-thirds of those who take the drug experience one or more side-effects. It has been hypothesized that the GI microbiota may protect from or contribute to these GI disturbances. Using in vitro batch culture and human gut model systems, studies were conducted to determine whether increased availability of dietary lipids and/or orlistat affect the composition and/or activity of the faecal microbiota. Results from 24-h batch culture fermentation experiments demonstrated no effect of orlistat in the presence or absence of a dietary lipid (olive oil) on the composition of bacterial communities [as determined by fluorescence in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE) analyses], but did show there was great variability in the lipolytic activities of the microbiotas of individuals, as determined by gas chromatography analysis of long-chain fatty acids in samples. Subsequent studies focused on the effect of orlistat in the presence and absence of lipid in in vitro human gut model systems. Systems were run for 14 days with gut model medium (GMM) only (to steady state, SS), then fed at 12-h intervals with 50 mg orlistat, 2 g olive oil or a mixture of both for 14 days. FISH and DGGE were used to monitor changes in bacterial populations. Bacteria were cultivated from the GMM only (control) systems at SS. All strains isolated were screened for lipolytic activity using tributyrin agar. FISH and DGGE demonstrated that none of the compounds (singly or in combination) added to the systems had any notable effect on microbial population dynamics for any of the donors, although Subdoligranulum populations appeared to be inhibited by orlistat in the presence or absence of lipid. Orlistat had little or no effect on the metabolism of indigenous and added lipids in the fermentation systems, but there was great variability in the way the faecal microbiotas of the donors were able to degrade added lipids. Variability in lipid degradation could be correlated with the number and activity of isolated lipolytic bacteria. The mechanism by which orlistat and the GI microbiota cause side-effects in individuals is unknown, but several hypotheses have been proposed to account for their manifestation. The demonstration of great variability in the lipolytic activity of microbiotas to degrade lipids led to a large-scale cultivation-based study of lipolytic/lipase-positive bacteria present in the human faecal microbiota. Of 4,000 colonies isolated from 15 donors using five different agars, 378 strains were identified that had lipase activity. Molecular identification of strains isolated from five donors demonstrated that lipase activity is more prevalent in the human GI microbiota than previously thought, with members of the phyla Firmicutes, Bacteroidetes and Actinobacteria identified. Molecular identification and characterization of the substrate specificities of the strains will be carried out as part of ongoing work.

Influence of substrate on corneal epithelial cell viability within ocular surface models

Corneal tissue engineering has improved dramatically over recent years. It is now possible to apply these technological advancements to the development of superior in vitro ocular surface models to reduce animal testing. We aim to show the effect different substrates can have on the viability of expanded corneal epithelial cells and that those which more accurately mimic the stromal surface provide the most protection against toxic assault. Compressed collagen gel as a substrate for the expansion of a human epithelial cell line was compared against two well-known substrates for modeling the ocular surface (polycarbonate membrane and conventional collagen gel). Cells were expanded over 10 days at which point cell stratification, cell number and expression of junctional proteins were assessed by electron microscopy, immunohistochemistry and RT-PCR. The effect of increasing concentrations of sodium lauryl sulphate on epithelial cell viability was quantified by MTT assay. Results showed improvement in terms of stratification, cell number and tight junction expression in human epithelial cells expanded upon either the polycarbonate membrane or compressed collagen gel when compared to a the use of a conventional collagen gel. However, cell viability was significantly higher in cells expanded upon the compressed collagen gel. We conclude that the more naturalistic composition and mechanical properties of compressed collagen gels produces a more robust corneal model.

On the performance of the tick test

In financial research, the sign of a trade (or identity of trade aggressor) is not always available in the transaction dataset and it can be estimated using a simple set of rules called the tick test. In this paper we investigate the accuracy of the tick test from an analytical perspective by providing a closed formula for the performance of the prediction algorithm. By analyzing the derived equation, we provide formal arguments for the use of the tick test by proving that it is bounded to perform better than chance (50/50) and that the set of rules from the tick test provides an unbiased estimator of the trade signs. On the empirical side of the research, we compare the values from the analytical formula against the empirical performance of the tick test for fifteen heavily traded stocks in the Brazilian equity market. The results show that the formula is quite realistic in assessing the accuracy of the prediction algorithm in a real data situation.

The effects of distraction on metacognition and metacognition on distraction: evidence from recognition memory

The effects of auditory distraction in memory tasks have been examined to date with procedures that minimize participants’ control over their own memory processes. Surprisingly little attention has been paid to metacognitive control factors which might affect memory performance. In this study, we investigate the effects of auditory distraction on metacognitive control of memory, examining the effects of auditory distraction in recognition tasks utilizing the metacognitive framework of Koriat and Goldsmith (1996), to determine whether strategic regulation of memory accuracy is impacted by auditory distraction. Results replicated previous findings in showing that auditory distraction impairs memory performance in tasks minimizing participants’ metacognitive control (forced-report test). However, the results revealed also that when metacognitive control is allowed (free-report tests), auditory distraction impacts upon a range of metacognitive indices. In the present study, auditory distraction undermined accuracy of metacognitive monitoring (resolution), reduced confidence in responses provided and, correspondingly, increased participants’ propensity to withhold responses in free-report recognition. Crucially, changes in metacognitive processes were related to impairment in free-report recognition performance, as the use of the ‘don’t know’ option under distraction led to a reduction in the number of correct responses volunteered in free-report tests. Overall, the present results show how auditory distraction exerts its influence on memory performance via both memory and metamemory processes.

Occurrence probability, width and number of steps of cusp precipitation for fully pulsed reconnection at the dayside magnetopause

We discuss the characteristics of magnetosheath plasma precipitation in the “cusp” ionosphere for when the reconnection at the dayside magnetopause takes place only in a series of pulses. It is shown that even in this special case, the low-altitude cusp precipitation is continuous, unless the intervals between the pulses are longer than observed intervals between magnetopause flux transfer event (FTE) signatures. We use FTE observation statistics to predict, for this case of entirely pulsed reconnection, the occurrence frequency, the distribution of latitudinal widths, and the number of ion dispersion steps of the cusp precipitation for a variety of locations of the reconnection site and a range of values of the local de-Hoffman Teller velocity. It is found that the cusp occurrence frequency is comparable with observed values for virtually all possible locations of the reconnection site. The distribution of cusp width is also comparable with observations and is shown to be largely dependent on the distribution of the mean reconnection rate, but pulsing the reconnection does very slightly increase the width of that distribution compared with the steady state case. We conclude that neither cusp occurrence probability nor width can be used to evaluate the relative occurrence of reconnection behaviors that are entirely pulsed, pulsed but continuous and quasi-steady. We show that the best test of the relative frequency of these three types of reconnection is to survey the distribution of steps in the cusp ion dispersion characteristics.

The accuracy of Doppler radar wind retrievals using insects as targets

Insect returns from the UK's Doppler weather radars were collected in the summers of 2007 and 2008, to ascertain their usefulness in providing information about boundary layer winds. Such observations could be assimilated into numerical weather prediction models to improve forecasts of convective showers before precipitation begins. Significant numbers of insect returns were observed during daylight hours on a number of days through this period, when they were detected at up to 30 km range from the radars, and up to 2 km above sea level. The range of detectable insect returns was found to vary with time of year and temperature. There was also a very weak correlation with wind speed and direction. Use of a dual-polarized radar revealed that the insects did not orient themselves at random, but showed distinct evidence of common orientation on several days, sometimes at an angle to their direction of travel. Observation minus model background residuals of wind profiles showed greater bias and standard deviation than that of other wind measurement types, which may be due to the insects' headings/airspeeds and to imperfect data extraction. The method used here, similar to the Met Office's procedure for extracting precipitation returns, requires further development as clutter contamination remained one of the largest error contributors. Wind observations derived from the insect returns would then be useful for data assimilation applications.

Influence of time on the plant availability of Cd, Ni and Zn after sewage sludge has been applied to soils

A pot experiment was conducted to test the hypothesis that decomposition of organic matter in sewage sludge and the consequent formation of dissolved organic compounds (DOC) would lead to an increase in the bioavailability of the heavy metals. Two Brown Earth soils, one with clayey loam texture (CL) and the other a loamy sand (LS) were mixed with sewage sludge at rates equivalent to 0, 10 and 50 1 dry sludge ha(-1) and the pots were sown with ryegrass (Lolium perenne L.). The organic matter content and heavy metal availability assessed with soil extractions with 0.05 M CaCl2 were monitored over a residual time of two years, while plant uptake over one year, after addition of the sludge. It was found that the concentrations of Cd and Ni in both the ryegrass and the soil extracts increased slightly but significantly during the first year. In most cases, this increase was most evident especially at the higher sludge application rate (50 t ha(-1)). However, in the second year metal availability reached a plateau. Zinc concentrations in the ryegrass did not show an increase but the CaCl2 extracts increased during the first year. In contrast, organic matter content decreased rapidly in the first months of the first year and much more slowly in the second (total decrease of 16%). The concentrations of DOC increased significantly in the more organic rich CL soil in the course of two years. The pattern followed by the decomposition of organic matter with time and the production of DOC may provide at least a partial explanation for trend towards increased metal availability.