E. I. Duff

Alkyl phenols and diethylhexyl phthalate in tissues of sheep grazing pastures fertilized with sewage sludge or inorganic fertilizer.

We studied selected tissues from ewes and their lambs that were grazing pastures fertilized with either sewage sludge (treated) or inorganic fertilizer (control) and determined concentrations of alkylphenols and phthalates in these tissues. Mean tissue concentrations of alkylphenols were relatively low (< 10–400 μg/kg) in all animals and tissues. Phthalates were detected in tissues of both control and treated animals at relatively high concentrations (> 20,000 μg/kg in many tissue samples). The use of sludge as a fertilizer was not associated with consistently increased concentrations of either alkylphenols or phthalates in the tissues of animals grazing treated pastures relative to levels in control animal tissues. Concentrations of the two classes of chemicals differed but were of a similar order of magnitude in liver and muscle as well as in fat. Concentrations of each class of compound were broadly similar in tissues derived from ewes and lambs. Although there were significant differences (p < 0.01 or p < 0.001) between years (cohorts) in mean tissue concentrations of both nonylphenol (NP) and phthalate in each of the tissues from both ewes and lambs, the differences were not attributable to either the age (6 months or 5 years) of the animal or the duration of exposure to treatments. Octylphenol concentrations were generally undetectable. There was no consistent cumulative outcome of prolonged exposure on the tissue concentrations of either class of pollutant in any ewe tissue. Mean tissue concentrations of phthalate were higher (p < 0.001) in the liver and kidney fat of male compared with female lambs. We suggest that the addition of sewage sludge to pasture is unlikely to cause large increases in tissue concentrations of NP and phthalates in sheep and other animals with broadly similar diets and digestive systems (i.e., domestic ruminants) grazing such pasture.

Phthalate and alkyl phenol concentrations in soil following applications of inorganic fertiliser or sewage sludge to pasture and potential rates of ingestion by grazing ruminants

Soil concentrations of dioctyl phthalate (DOP) and the alkyl phenols, octyl phenol (OP) and nonyl phenol (NP), after repeated surface applications of sewage sludge to pastures, were investigated. Liquid sludge was applied at a rate of 2.25 tonnes dry matter (DM) per hectare to each of three treated (T) plots on three occasions during the summer and two occasions in the early spring over a period of 2.5 years. Control (C) plots were treated with inorganic fertiliser containing amounts of nitrogen equivalent to those applied to the treated plots. At between 69 and 81 days after the application of sludge, 15 separate soil samples were collected from one half of each of the plots (Experiment 1). Concentrations (microg g(-1)) of DOP were higher (P < 0.001) than those of NP, while those of OP were generally below detectable levels. Mean soil concentrations of DOP were not significantly different in T and C plots [0.233 vs. 0.155 microg g(-1); standard error of the difference (SED) = 0.046; not significant (NS)], partly because there was already a relatively large amount of DOP present. NP concentrations were, however, significantly higher in T than in C plots (0.021 vs. 0.013 microg g(-1) SED = 0.002; P < 0.05). There was no consistent change over time in the mean soil concentrations of these compounds when sampled at intervals of 3-6 months. Concentrations in soil samples collected at monthly intervals following sludge application indicated that the variation in concentrations of these endocrine-disrupting compounds (EDC) was unrelated to time since sludge application. Rates of soil ingestion, expressed as the percentage of DM intake represented by soil, were higher during the winter than the summer (5.40 vs. 1.17; SED = 0.360; P < 0.001) and estimated daily intakes of DOP and NP were up to 150 microg and 8 microg, respectively. It is concluded that the application of sewage sludge to pasture does not increase soil concentrations of phthalate (as DOP) or alkyl phenols. Thus, the risk of increased exposure to these EDC as a result of sludge application is small. However, the small effect of sludge application on soil concentrations may be largely a reflection of the relatively high concentrations of DOP already present in the soil, which may be biologically significant.

Maternal and fetal tissue accumulation of selected endocrine disrupting compounds (EDCs) following exposure to sewage sludge-treated pastures before or after conception

This paper reports patterns of accumulation of selected EDCs in adult and fetal livers of sheep exposed to sewage sludge-treated pastures, at different times and at two stages of development.

Concentrations and geographic distribution of selected organic pollutants in Scottish surface soils.

Concentrations of selected persistent organic pollutants (POPs) representing three chemical classes (polycyclic aromatic hydrocarbons (PAH), polybrominated diphenyl ethers (PBDE) and polychlorinated biphenyls (PCB) and the organic pollutant diethylhexyl phthalate (DEHP), were determined in surface soil samples (0-5 cm) collected at 20 km grid intersects throughout Scotland over a three-year period. Detectable amounts of all chemical classes and most individual congeners were present in all samples. There were no consistent effects of soil or vegetation type, soil carbon content, pH, altitude or distance from centres of population on concentrations which exhibited extreme variation, even in adjacent samples. It is concluded that soil POPs and DEHP concentrations and associated rates of animal and human exposure were highly variable, influenced by multiple, interacting factors, and not clearly related to local sources but possibly related to wet atmospheric deposition and the organic carbon content of the soil.

Effect of long-term feed restriction on seasonal endocrine changes in Soay sheep

Groups of 15 adult, castrated, male Soay sheep were housed under natural daylength conditions at 57 degrees N and fed a complete diet ad libitum (AL) or at a restricted rate (R) of 35 g dry matter (DM)/kg(0.75) initial liveweight per day. The diet was based on barley and dried grass pellets and contained an estimated 11.6 MJ of metabolisable energy, 83% DM and 140 g crude protein/kg DM. In the AL animals, higher levels of feed intake during the periods of long daylength were associated with shorter inter-meal intervals (p<0. 001), a greater meal frequency (p<0.001), and a greater proportion of time spent eating (p<0.001) together with a greater rate of feed ingestion (p<0.001) and an increased meal size (p<0.001). Mean plasma concentrations of insulin, prolactin, insulin-like growth factor 1 (IGF-1), triiodothyronine (T(3)), and thyroxine (T(4)) were higher (p<0.001) in the spring or summer than in the autumn. Mean plasma GH concentrations did not differ with month. Compared with R animals, AL animals had higher mean plasma concentrations of insulin (p<0.001), prolactin (p<0.01), T(3) (p<0.01), and T(4) (p<0.01). Plasma GH and IGF-1 concentrations did not differ significantly with treatment. There was a greater increase in plasma insulin concentrations following feeding in R than AL animals (p<0.001) owing to higher pre-feeding concentrations in AL animals and the ingestion of larger amounts of feed by R than AL animals in the period after fresh feed was introduced. There were significant differences between months in this response, in R animals (p<0.01). Mean CSF insulin concentrations were significantly higher in AL than R animals (p<0.05) but were not affected by month. Neither was there a difference between pre-feeding concentrations and concentrations at approximately 12 h after feeding. It is concluded that the differences in the response of plasma insulin concentrations to feeding at different times of year, which were detected in R animals, were attributable, primarily, to differences in the vagally-induced insulin response to feeding and that these differences may provide important feedback signals to the appetite centre.

Small genetic differences between ericoid mycorrhizal fungi affect nitrogen uptake by Vaccinium

Ericoid mycorrhizal fungi have been shown to differ in their pattern of nitrogen (N) use in pure culture. Here, we investigate whether this functional variation is maintained in symbiosis using three ascomycetes from a clade not previously shown to include ericoid mycorrhizal taxa. Vaccinium macrocarpon and Vaccinium vitis-idaea were inoculated with three fungal strains known to form coils in Vaccinium roots, which differed in their patterns of N use in liquid culture. (15)N was used to trace the uptake of -N, -N and glutamine-N into shoots. (15)N transfer differed among the three fungal strains, including two that had identical internal transcribed spacer (ITS) sequences, and was quantitatively related to fungal growth in liquid culture at low carbon availability. These results demonstrate that functional differences among closely related ericoid mycorrhizal fungi are maintained in symbiosis with their hosts, and suggest that N transfer to plant shoots in ericoid mycorrhizas is under fungal control.

A study on temporal trends and estimates of fate of Bisphenol A in agricultural soils after sewage sludge amendment

Temporal concentration trends of BPA in soils were investigated following sewage sludge application to pasture (study 1: short term sludge application; study 2: long term multiple applications over 13 years). The background levels of BPA in control soils were similar, ranging between 0.67-10.57 ng g(-1) (mean: 3.02 ng g(-1)) and 0.51-6.58 ng g(-1) (mean: 3.22 ng g(-1)) for studies 1 and 2, respectively. Concentrations in both treated and control plots increased over the earlier sampling times of the study to a maximum and then decreased over later sampling times, suggesting other sources of BPA to both the treated and control soils over the study period. In study 1 there was a significant treatment effect of sludge application in the autumn (p=0.002) although no significant difference was observed between treatment and control soils in the spring. In study 2 treated soils contained considerably higher BPA concentrations than controls ranging between 12.89-167.9 ng g(-1) (mean: 63.15 ng g(-1)). This and earlier studies indicate the long-term accumulation of multiple contaminants by multiple sewage sludge applications over a prolonged period although the effects of the presence of such contaminant mixtures have not yet been elucidated. Fugacity modelling was undertaken to estimate partitioning of Bisphenol A (soil plus sewage: pore water: soil air partitioning) and potential uptake into a range of food crops. While Bisphenol A sorbs strongly to the sewage-amended soil, 4% by mass was predicted to enter soil pore water resulting in significant uptake by crops particularly leafy vegetables (3.12-75.5 ng g(-1)), but also for root crops (1.28-31.0 ng g(-1)) with much lower uptake into cereal grains (0.62-15.0 ng g(-1)). This work forms part of a larger programme of research aimed at assessing the risks associated with the long-term application of sewage sludge to agricultural soils.

Temporal changes in soil properties at an upland Scottish site between 1956 and 1997

The aim of this study was to examine the frequency with which soil samples require to be taken in order to determine significant temporal changes in soil properties. The examination was carried out using data from Glensaugh Research Station in north-east Scotland where podzolic soils were sampled in 1956, 1977 and 1997, and by re-analysis of archived material. Significant differences in chemistry due to storage were detected, particularly decreases in pH of air-dried organic soils. In these cases original data were used for statistical analysis to establish changes between 1956 and 1997. Temporal changes were found for exchangeable Ca and Mg which generally decreased with time throughout the soil profile, whereas exchangeable H increased. Derived data, such as percent base saturation, declined dramatically due to decreases in exchangeable base cations. Similar podzolic soils were sampled at an adjacent Environmental Change Network (ECN) site in 1993. Application of statistical techniques to the ECN soil chemistry data allowed an estimation of the detectable change between any two years. These data along with the rates of temporal change from 1956 to 1997 allowed the calculation of the number of years required for measurable changes to be achieved. These changes and sampling intervals vary among different horizons and chemistries. Although they are site-specific, they do confirm that the current ECN protocols of a 5-year and 20-year sampling would be appropriate in order to detect changes in soil properties over time at this site.

Impact of Sublethal Levels of Environmental Pollutants Found in Sewage Sludge on a Novel Caenorhabditis elegans Model Biosensor

A transgenic strain of the model nematode Caenorhabditis elegans in which bioluminescence reports on relative, whole-organism ATP levels was used to test an environmentally-relevant mixture of pollutants extracted from processed sewage sludge. Changes in bioluminescence, following exposure to sewage sludge extract, were used to assess relative ATP levels and overall metabolic health. Reproductive function and longevity were also monitored. A short (up to 8 h) sublethal exposure of L4 larval stage worms to sewage sludge extract had a concentration-dependent, detrimental effect on energy status, with bioluminescence decreasing to 50–60% of the solvent control (1% DMSO). Following longer exposure (22–24 h), the energy status of the nematodes showed recovery as assessed by bioluminescence. Continuous exposure to sewage sludge extract from the L4 stage resulted in a shorter median lifespan relative to that of solvent or medium control animals, but only in the presence of 400–600 µM 5-fluoro-2′-deoxyuridine (FUdR), which was incorporated to inhibit reproduction. This indicated that FUdR increased lifespan, and that the effect was counteracted by SSE. Exposure to sewage sludge extract from the L1 stage led to slower growth and a delayed onset of egg laying. When L1 exposed nematodes reached the reproductive stage, no effect on egg laying rate or egg number in the uterus was observed. DMSO itself (1%) had a significant inhibitory effect on growth and development of C. elegans exposed from the L1 stage and on reproduction when exposed from the L4 stage. Results demonstrate subtle adverse effects on C. elegans of a complex mixture of environmental pollutants that are present, individually, in very low concentrations and indicate that our biosensor of energy status is a novel, sensitive, rapid, quantitative, whole-organism test system which is suitable for high throughput risk assessment of complex pollutant mixtures.

Geographic variation in tissue accumulation of endocrine disrupting compounds (EDCs) in grazing sheep.

Muscle tissue was collected from ewes and lambs derived from farms throughout Scotland and sample concentrations of five endocrine disrupting compound groups were determined. Farms of origin were categorised according to geographic region. There were few statistically-significant differences with region or distance from cities. However, the magnitude of the difference between the highest and lowest mean values in ewe muscle from different regions exceeded 30% for 13 of the 15 compounds that were consistently detected in muscle, with animals derived from the industrialised region having the highest mean values for 11 of the 13 compounds. A less marked trend was apparent in the lamb muscle (8 of 13 highest were in the industrialised region). The physiological effects of such small differences in exposure to mixtures of pollutants remain to be determined.