Presence of Muscarinic Acetylcholine Receptors in the Cattle Tick Boophilus microplus and in Epithelial Tissue Culture Cells of Chironomus tentans

A muscarinic acetylcholine receptor (mAChR) has been demonstrated and partially characterized in larvae of the cattle tick Boophilus microplus. Its properties are compared with mAChR from an epithelial cell line from the dipteran insect Chironomus tentans. Competition studies with cholinergic ligands of different specificity revealed the muscarinic nature of the cholinergic receptors investigated in both species. In homogenates from tick larvae, specific binding sites for [³H]quinuclidinyl benzilate (QNB) with high affinity (1·2±(0·13) nM ; B_max 22·5 pmol mg protein⁻¹) were detected that do not bind nicotinic compounds specifically. The estimated IC₅₀ values for nicotine, imidacloprid and α-bungarotoxin were all in the mM range. Additionally, with tick larvae, high-affinity nicotinic binding sites were detected with [³H]nicotine which could be displaced by high concentrations of imidacloprid or QNB. The estimated IC₅₀ values for nicotine, α-bungarotoxin, imidacloprid and QNB were 43(±8) nM , 0·8(±0·2) μM , 2·8(±0·6) μM and 78(±1·9) μM , respectively. With homogenates of the non-neuronal insect cell line from C. tentans, only high-affinity binding sites for [³H]QNB were found. Muscarinic antagonists selectively displaced [³H]quinuclidinyl benzilate (QNB) binding to tick larvae homogenates. The mAChR of B. microplus preferred pirenzepine (IC₅₀ 2·13(±1·02) μM ) among different subtype-specific mAChR antagonists (4-DAMP had IC₅₀ 49·9(±9·13) μM and methoctramine had IC₅₀ 121(±14·2) μM ) indicating a type of binding site similar to the vertebrate M1 mAChR subtype. The tick muscarinic receptor seems to be a G-protein-coupled receptor, as concluded from the 4·8-fold reduction in receptor affinity for binding of the muscarinic agonist oxotremorine M upon treatment with the non-hydrolysable GTP-analogue γ-S-GTP. Binding data for the agonists oxotremorine M (IC₅₀ 71·3(±19·6) μM ) and carbachol (IC₅₀ 253(±87·1) μM ) parallel the biological efficacy of these compounds, in that, while oxotremorine M showed some activity against ticks, carbachol was ineffective.     

Spatial pattern of trees influences species productivity in a mature oak–pine mixed forest

Spatial pattern has a key role in the interactions between species in plant communities. These interactions influence ecological processes involved in the species dynamics: growth, regeneration and mortality. In this study, we investigated the effect of spatial pattern on productivity in mature mixed forests of sessile oak and Scots pine. We simulated tree locations with point process models and tree growth with spatially explicit individual growth models. The point process models and growth models were fitted with field data from the same stands. We compared species productivity obtained in two types of mixture: a patchy mixture and an intimate mixture. Our results show that the productivity of both species is higher in an intimate mixture than in a patchy mixture. Productivity difference between the two types of mixture was 11.3 % for pine and 14.7 % for oak. Both species were favored in the intimate mixture because, for both, intraspecific competition was more severe than interspecific competition. Our results clearly support favoring intimate mixtures in mature oak–pine stands to optimize tree species productivity; oak is the species that benefits the most from this type of management. Our work also shows that models and simulations can provide interesting results for complex forests with mixtures, results that would be difficult to obtain through experimentation.     

Indicative properties of fly‐ash affected forest soils in Northeastern Germany

Many forest ecosystems in Germany are strongly influenced by emissions of pollutants like SO₂ and alkaline dusts. To quantify and evaluate the consequences of long‐term fly ash deposition on forest soils, a study was conducted in pine stands (Pinus sylvestris) in the Dübener Heide in Northeastern Germany. This forest area has been influenced mainly by emissions from coal‐fired power plants and the chemical industry of the industrial region Bitterfeld‐Wolfen‐Zschornewitz since the early 1900. The study sites are located along a fly ash deposition gradient of 8, 16, 14, 18, and 25 km away from the main emission source in Bitterfeld (sites 1, 2, 3, 4, and 5, respectively). Samples of the organic horizons (Oi, Oe, and Oa) and mineral topsoil (0—10 cm) were taken in fall 1998 and analyzed for their ferromagnetic susceptibility and total ash content. Scanning electron microscopy (SEM) and energy dispersive X‐ray microanalysis (EDX) were performed on selected samples to differentiate between the pedogenic and atmospheric origin of the mineral components in the organic horizons. As a result of the long‐term deposition, ferromagnetic fly ash components are mainly accumulated in the Oe and Oa horizons of the forest soils studied. Ferromagnetic susceptibility was significantly higher (p ≤ 0.05) in the Oe horizon of sites 1 and 2 compared to sites 3, 4, and 5. Unusually high total ash contents for organic horizons of > 74 % were determined in the Oa at all sites. SEM revealed 3 distinct features of persistent fly ash deposits from coal‐fired power plants within the organic horizons that can be defined as ”︁stable glasses” with magnetic properties, aluminum‐silicate‐minerals, and slag fragments. SEM and EDX indicated that a great portion of the mineral particles found in the organic horizons of forests soils influenced by fly ash are from atmospheric sources. For detection of atmospheric lignite‐derived deposition into forest soils, the Oe and Oa horizons have to be considered as specific diagnostic horizons because they show indicative properties for such soils.     

Organic‐carbon fractions in an agricultural topsoil assessed by the determination of the soil mineral surface area

According to recent conceptual models, the organic carbon (OC) of soils can be divided into OC fractions of increasing stability from labile free OC to resistant OC associated with the soil mineral phase. In this study, we present a method for quantifying two OC fractions based on soil aggregate–size fractionation and the N₂ gas–adsorption method. For this purpose, we analyzed soil material of the plow layer of a Haplic Chernozem subjected to different fertilizer treatments (no fertilizer, mineral fertilizer, mineral and organic fertilizer). The total organic‐C concentration (TOC) and the clay content of the different size fractions were determined as well as the specific surface area (SSA_mineral) and the sample pore volume after thermal oxidation (OC‐free). The TOC of the different soil‐aggregate fractions was linearly related to SSA_mineral. Clay‐associated OC and nonassociated OC fractions of the different soil samples were quantified using two methods based on the OC surface loading at the clay fraction. The application of organic fertilizer increased the amount of nonassociated OC but hardly affected the concentration of clay‐associated OC. This finding agrees with previous studies on C dynamics in soils and indicates a finite capacity of soil materials to sequester OC. Even without any addition of organic fertilizer, the mineral phase of the analyzed soil material appears to be C‐saturated.     

Mobile and readily available C and N fractions and their relationship to microbial biomass and selected enzyme activities in a sandy soil under different management systems

Within different land‐use systems such as agriculture, forestry, and fallow, the different morphology and physiology of the plants, together with their specific management, lead to a system‐typical set of ecological conditions in the soil. The response of total, mobile, and easily available C and N fractions, microbial biomass, and enzyme activities involved in C and N cycling to different soil management was investigated in a sandy soil at a field study at Riesa, Northeastern Germany. The management systems included agricultural management (AM), succession fallow (SF), and forest management (FM). Samples of the mineral soil (0—5, 5—10, and 10—30 cm) were taken in spring 1999 and analyzed for their contents on organic C, total N, NH₄⁺‐N and NO₃^—‐N, KCl‐extractable organic C and N fractions (Corg_(KCl) and Norg_(KCl)), microbial biomass C and N, and activities of β‐glucosidase and L‐asparaginase. With the exception of Norg_(KCl), all investigated C and N pools showed a clear relationship to the land‐use system that was most pronounced in the 0—5 cm profile increment. SF resulted in greater contents of readily available C (Corg_(KCl)), NH₄⁺‐N, microbial biomass C and N, and enzyme activities in the uppermost 5 cm of the soil compared to all other systems studied. These differences were significant at P ≤ 0.05 to P ≤ 0.001. Comparably high C_mic:C_org ratios of 2.4 to 3.9 % in the SF plot imply a faster C and N turnover than in AM and FM plots. Forest management led to 1.5‐ to 2‐fold larger organic C contents compared to SF and AM plots, respectively. High organic C contents were coupled with low microbial biomass C (78 μg g^—1) and N contents (10.7 μg g^—1), extremely low C_mic : C_org ratios (0.2—0.6 %) and low β‐glucosidase (81 μg PN g^—1 h^—1) and L‐asparaginase (7.3 μg NH₄‐N g^—1 2 h^—1) activities. These results indicate a severe inhibition of mineralization processes in soils under locust stands. Under agricultural management, chemical and biological parameters expressed medium values with exception for NO₃^—‐N contents which were significantly higher than in SF and FM plots (P ≤ 0.005) and increased with increasing soil depth. Nevertheless, the depth gradient found for all studied parameters was most pronounced in soils under SF. Microbial biomass C and N were correlated to β‐glucosidase and L‐asparaginase activity (r ≥ 0.63; P ≤ 0.001). Furthermore, microbial biomass and enzyme activities were related to the amounts of readily mineralizable organic C (i.e. Corg_(KCl)) with r ≥ 0.41 (P ≤ 0.01), suggesting that (1) KCl‐extractable organic C compounds from field‐fresh prepared soils represent an important C source for soil microbial populations, and (2) that microbial biomass is an important source for enzymes in soil. The Norg_(KCl) pool is not necessarily related to the size of microbial biomass C and N and enzyme activities in soil.<?show $6#>     

Ammonium‐driven nitrification plays a key role in increasing Mn availability in calcareous soils

Background : Manganese deficiency often becomes a yield limiting factor, particularly on calcareous soils, even though the total soil manganese content is usually sufficient. Although it is known that acidifying N fertilizers can improve Mn availability, the reason of this effect is still unknown. Aim : Our aim was to investigate the effect of stabilized ammonium fertilizers as a tool to distinguish between physiological‐ and nitrification‐induced acidification. Method : Two pot experiments with Triticum aestivum L. and one soil incubation experiment using different nitrogen forms (CN = calcium nitrate, AN = ammonium nitrate, AS = ammonium sulfate, ATS = ammonium thiosulfate) with and without addition of nitrification inhibitors (DCD, Nitrapyrin, Piadin, DMPP) were conducted to examine the effect on Mn availability in the soil and Mn uptake by the plants at different development stages (EC 31 und 39). Results : With increasing fertilizer ${\mathrm{NH}}_4^{+}$ content a higher Mn concentration was detected: CN: 32 µg Mn g^?1 DM, AN: 39 µg Mn g^?1 DW, AS: 55 µg Mn g^?1 DM, ATS: 109 µg Mn g^?1 DM. The addition of a nitrification inhibitor resulted in a significantly lower rhizosphere pH compared to the non‐stabilized fertilizer. Surprisingly, the use of different nitrification inhibitors led to unchanged (CN, AN) or lower Mn concentrations of wheat. Especially in the ${\mathrm{NH}}_4^{+}$ treatments (AS and ATS), this negative effect was very evident (AS+DCD: 42 µg Mn g^?1 DM; ATS+DCD: 55 µg Mn g^?1 DM). Conclusions : Mn availability was enhanced by ongoing nitrification process rather than physiological acidification. Compared to other N forms, ammonium thiosulfate led to the highest Mn availability in bulk soil.     

Few alterations in clinical pathology and histopathology observed in a CYP2C18&19 humanized mice model

Background

This study was performed to characterize a gene-addition transgenic mouse containing a BAC (bacterial artificial chromosome) clone spanning the human CYP2C18&19 genes (tg-CYP2C18&19).

Methods

Hemizygous tg-CYP2C18&19, 11 week old mice were compared with wild-type littermates to obtain information regarding clinical status, clinical pathology and anatomical pathology. After one week of clinical observations, blood samples were collected, organs weighed, and tissues collected for histopathology.

Results

In males, the tissue weights were lower in tg-CYP2C18&19 than in wild-type mice for brain (p ≤ 0.05), adrenal glands (p ≤ 0.05) and brown fat deposits (p ≤ 0.001) while the heart weight was higher (p ≤ 0.001). In female tg-CYP2C18&19, the tissue weights were lower for brain (p ≤ 0.001) and spleen (p ≤ 0.001) compared to wild-type females. Male tg-CYP2C18&19 had increased blood glucose levels (p ≤ 0.01) while females had decreased blood triglyceride levels (p ≤ 0.01).

Conclusion

Despite the observed alterations, tg-CYP2C18&19 did not show any macroscopic or microscopic pathology at the examined age. Hence, these hemizygous transgenic mice were considered to be viable and healthy animals.