Isolation and characterisation of symbiotically effective Rhizobium resistant to arsenic and heavy metals after the toxic spill at the Aznalcóllar pyrite mine

After the toxic spill occurred at Aznalcóllar pyrite mine (Southern Spain), a wide area of croplands near the Doñana Wild Park was contaminated with 4.5 million m³ of slurries composed of acidic waters loaded with toxic metals and metalloids such as As, Sb, Zn, Pb, Cu, Co, Tl, Bi, Cd, Ag, Hg and Se. Today, 6 years after the spill, the concentration of toxic elements in these soils is still very high, in spite of the efforts to clean the zone. However, some plant species have colonised this contaminated area. Legumes possessing N₂-fixing nodules on their roots represented a significant proportion of these plants. Our objective was to use the Rhizobium-legume symbiosis as a new tool for bioremediate the affected area. We have isolated about 100 Rhizobium strains, 41 of them being resistant to high concentrations of As (300 mg l⁻¹), Cu (100 mg l⁻¹) and Pb (500 mg l⁻¹). Their phenotypes and bioaccumulation potentials have been characterised by their growth rates in media supplemented with As and heavy metals. The presence of the resistance genes in some strains has been confirmed by PCR and Southern blot hybridisation. Several Rhizobium were symbiotically effective in the contaminated soils. On the other hand, the first steps in nodule establishment seemed to be more affected by heavy metals than N₂-fixation.     

Technical and Environmental Functioning of Detention Ponds for the Treatment of Highway and Road Runoff

Based on an inventory of more than 200 facilities for the treatment of road runoff water in Sweden, 26 representative stormwater ponds were studied with regard to technical function, maintenance and metal accumulation. Sediment accumulation and metal content in the pond sediment was studied in relation to concentration in the runoff water, spatial variation and pond geometry. It emerged that the pollution retention capacity was not optimal for many of the ponds. The observed sediment thickness was low reflecting turbulent conditions and that much of the pollutants are transported through the ponds. Statistical tests performed on 15 ponds showed a significant relationship between metal content in sediment and pond size while pond shape was found to have no effect on accumulation rate. There is still significant potential for the development of the design and technical function of the ponds, e.g. design elements such as pre-settlement trench and elements for enhancing high hydraulic efficiency. Disturbance of the technical function, in terms of clogging of pipes, erosion, flooding etc. was reviewed for three of the ponds, however only 50% of the ponds did show some form of documented maintenance-program. This generates an obvious threat to the effective long-term function of these ponds.     

Changes in soil fungal:bacterial biomass ratios following reductions in the intensity of management of an upland grassland

In this study we examined the effect on soil fungal:bacterial biomass ratios of withholding fertiliser, lime, and sheep-grazing from reseeded upland grassland. The cessation of fertiliser applications on limed and grazed grassland resulted in a reduction in soil pH from 5.4 to 5.1. The cessation of fertiliser applications and liming on grazed grassland resulted in a fall in pH from 5.4 to 4.7, whereas withholding fertiliser and lime and the removal of grazing resulted in a further reduction to pH 4.5. Substrate-induced respiration was reduced in the unfertilised grazed (21%; P<0.01) and unfertilised ungrazed (36%; P<0.001) treatments. Bacterial substrate-induced respiration and bacterial fatty acids were unaffected by the treatments. The relative abundance of the fungal fatty acid 18:26 increased by 39 and 72% (P<0.05) in the limed grazed and unfertilised grazed treatments, respectively. Fungal substrate-induced respiration increased in the limed grazed (18%) and unfertilised grazed (65%; P<0.05) treatments. The ratio of 18:26: bacterial fatty acids was correlated with the ratio of fungal:bacterial substrate-induced respiration (r=0.69; P<0.001).     

Clear-cutting affects the ammonia-oxidising community differently in limed and non-limed coniferous forest soils

The effects of clear-cutting on the ammonia-oxidising bacterial community were studied in the soil of limed and non-limed spruce forest plots located in the central part of southern Sweden. The communities were studied using denaturing gradient gel electrophoresis (DGGE) profiling after polymerase chain reaction (PCR) amplification from total DNA with primers reported to be specific for -subgroup ammonia-oxidising bacteria. The bands on the DGGE were sequenced and each unique sequence was interpreted as representing one ammonia-oxidising population. The relative abundance of each population was determined by measuring the fluorescence of the respective DGGE bands. In both limed and non-limed soil, the same two Nitrosospira populations were found, one belonging to cluster 2 (NScl2) and one to cluster 4 (NScl4). However, while NScl4 first appeared a year after the clear-cutting in the non-limed plot, it was present both before and after the cutting in the limed plot. Irrespective of previous liming, clear-cutting caused a shift in the ammonia-oxidiser community, from dominance by the NScl2 population to a community with approximately equal relative abundance of NScl2 and NScl4. In both plots the total size of the community increased after clear-cutting (based on increased DGGE band intensity), most likely due to increased NH₄⁺ availability, but the growth response was faster in the limed plot. Hence, the prior liming increased the responsiveness of the ammonia-oxidisers to the changes caused by cutting. This is the first study to report the effects of clear-cutting on the ammonia-oxidising community, and the results show a clear correlation between increased potential nitrification and a shift in the ammonia-oxidiser community.     

Soil Acidification Induced by Ammonium Sulphate Addition in a Norway Spruce Forest in Southwest Sweden

The contributions of different acidifying processes to the total protonload (TPL) of the soil in control plots (C) and ammonium sulphate treatedplots (NS) were studied in a Norway spruce stand in Southwest Sweden during 1988–1998. The annual deposition of inorganic nitrogen and sulphate was on average 18 kg N and 20 kg S ha^-1. In addition the NS treated plots received 100 kg N and 114 kg S ha^-1 annually. The amounts of nutrients added to the ecosystem by wet and dry deposition and the leaching at 50 cm depth were calculated. The net atmosphericproton load, the proton load by nitrogen transformations in the soil, the sulphate sorption/desorption in the soil and the excess base cation accumulation in biomass were calculated. There was no leaching of inorganic nitrogen from control plots during the study period. The net atmospheric proton deposition, originating from sulphuric and nitric acid deposition, was the main contributor to TPL in control plots. The addition of ammonium sulphate increased the leaching of ammonium, nitrate, sulphate, magnesium and calcium but not of potassium. The TPL in NS plots was about ten times that in control plots. The nitrogen transformation processes were the main contributors to TPL to NS soil, in the beginning by ammonium uptake and later also by nitrification. The pH decreased by 0.4 units in the mineral soil. The between-year variation in TPL during the eleven year period in C plots (200–1500 mol_c ha^-1 yr^-1) and in NS plots (1000–13000 mol_c ha^-1 yr^-1) was mainly dependent on the sorption or release of sulphate. Both in C and NS, the TPL was buffered mainly by dissolving solid aluminium compounds, most probably some Al(OH)₃ phase.     

Hydrogeochemistry of a stream draining sulfide-bearing postglacial sediments in Finland

The influence of hydrological conditions and overburden type on the stream-water quality in a catchment effected by fine-grained sulfide-bearing sediments was studied. Water samples for measurement of pH and electric conductivity were collected at 16 sites along the main course of the stream during autumn and spring high water flow and during low and intermediate water flow. On one occasion (spring high-water flow) samples for analysis of total organic carbon were collected at these sites. Seventeen water samples for analyses of chemical elements (ICP-MS and ICP-AES) were taken within the catchment basin during autumn high water flow. The pH and conductivity of snow, and the geochemistry of the sulfide sediments were also studied. The occurrence of fine-grained sediments within the catchment was estimated from topographic and geologic maps and by visit to areas were the overburden type could not be determined from the maps. The hydrological conditions had a strong influence on the water quality. During high water flow in spring and autumn the concentrations of ionic components (conductivity) and hydrogen ions in the stream water were high, while the concentrations were much lower during low and intermediate water flow and when the ground was frozen. Extensive leaching of the sulfide-bearing sediments, which had higher contents of S and metals than the fine fraction of till in the study area, resulted in elevated contents of Li, Ni, Zn, Co, B, Al, Mg, U, Sr, Ca, K and Na in the stream water during autumn high water flow. The contents of Cr, Ba, Pb, Si and Fe in stream water were not much influenced by the type of overburden, while V was depleted in drainage waters from the sulfide sediments. Humic substances flocculated in the water in the middle section of the stream, which may have been caused by di- and trivalent metal cations leached in large quantities from the oxidizing sulfide sediments.     

Occurrence and turnover of atmospheric mercury over the nordic countries

A seasonal variation of both particle and gaseous Hg concentrations in the atmosphere is present in south-western Sweden. An average gaseous Hg level of 3.7 ng m^?3 is found in winter, compared to 2.8 ng m^?3 in summer. A weak decreasing south-north gradient for gaseous Hg in air over the Nordic countries is also present, with yearly average values from 3.2 to 2.8 ng m^?3. A gradient for particulate Hg is less clear. An air parcel trajectory sector classification of gaseous Hg levels in air, and to some extent the particulate associated Hg, clearly demonstrates the increased concentrations in the southern sectors, especially in south-western Sweden where the gaseous Hg increase is about I ng m^?3. These observations are consistent with an influence from the European continent. The average concentrations of Hg in precipitation at the various stations show a pronounced decreasing south-north gradient. A major portion of the total Hg present in precipitation is associated with particles. For the southern stations, a strong correlation between Hg and sulfate, or pH, is present suggesting a connection between Hg in precipitation and anthropogenic activities.     

Cattle ticks in Malawi

Four genera of cattle ticks were found and identified during two tick surveys: Amblyomma, Boophilus, Hyalomma and Rhipicephalus. Amblyomma variegatum was the only species found of genus Amblyomma. The genus Boophilus was represented by two species: B. microplus and B. decoloratus, and the genus Hyalomma by two species: H. truncatum and H. marginatum rufipes. Ten different species of Rhipicephalus were identified: R. appendiculatus, R. sanguineus, R. compositus, R. pravus, R. simus, R. kochi, R. reichenowi, R. tricuspis, R. masseyi and R. punctatus.The results are compared with those from previous investigations conducted by the late Dr S.G. Wilson.All sites at which the different species of cattle ticks were found, and the distribution of each tick are illustrated by a series of maps. The territorial distribution of the ticks is discussed. R. appendiculatus was collected at five sites in the Shire Highlands, an area in which their presence has not been previously demonstrated.     

Ustekinumab pharmacokinetics after subcutaneous administration in swine model

BackgroundDue to multiple similarities in the structure and physiology of human and pig skin, the pig model is extremely useful for biological drug testing after subcutaneous administration. Knowledge of the differences between subcutaneous injection sites could have a significant impact on the absorption phase and pharmacokinetic profiles of biological drugs.ObjectivesThis study aimed to analyze the impact of administration site on pharmacokinetics and selected biochemical and hematological parameters after a single subcutaneous administration of ustekinumab in pigs. Drug concentrations in blood plasma were analyzed by enzyme-linked immunosorbent assay. Pharmacokinetic analyses were performed based on raw data using Phoenix WinNonlin 8.1 software and ThothPro v 4.1.MethodsThe study included 12 healthy, female, large white piglets. Each group received a single dose of ustekinumab given as a 1 mg/kg subcutaneous injection into the internal part of the inguinal fold or the external part of the inguinal fold.ResultsThe differences in absorption rate between the internal and external parts of the inguinal fold were not significant. However, the time of maximal concentration, clearance, area under the curve calculated between zero and mean residence time and mean residence time between groups were substantially different (p > 0.05). The relative bioavailability after administration of ustekinumab into the external part of the inguinal fold was 40.36% lower than after administration of ustekinumab into the internal part of the inguinal fold.ConclusionsHealthy breeding pigs are a relevant model to study the pharmacokinetic profile of subcutaneously administered ustekinumab.     

Testing Snow Mould Resistance of Winter Wheat: Inoculation Experiments with Microdochium nivale in the Field

Winter survival of 24 winter wheat lines was evaluated at three locations in south-eastern Norway in 1996-97 and 1997-98. Plants were inoculated with a mycelial suspension of Microdochium nivale or a preparation of M. nivale mycelium grown on boiled wheat grains, or they remained uninoculated. Artificially applied inoculum led to a higher disease pressure in some of the trials only. The effect of environmental conditions on variation in winter survival among wheat lines was greater than that of inoculation, and it was therefore difficult to draw a definite conclusion on the variation in specific snow mould resistance.