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Adaptation to a repeated restraint stress schedule was monitored in ethanol-treated and control rats. A single episode of 2 h restraint decreased food intake in both control and ethanol-treated rats. The decreases in control rats were not observed following the 5th daily restraint of 2 h/day, suggesting that adaptation has occurred. Ethanol-treated rats, however, exhibited decreased food intake even after 5th daily restraint of 2 h/day. Ethanol administration decreased weekly but not daily cumulative food intake in unrestrained rats. Food intakes of ethanol-treated and control restrained rats were comparable following 1st-3rd daily restraints, but were smaller in ethanol-treated rats following the 4th and 5th daily restraints. Open-field ambulatory activities monitored 24 h after the 5th daily restraint on the 6th day were comparable in control restrained and unrestrained rats. Ethanol-treated and control unrestrained rats also exhibited comparable ambulation, but ethanol-treated rats exhibited smaller activity than control restrained or ethanol-treated unrestrained rats. Fluid intakes of ethanol and control rats were comparable during the 2 weeks of ethanol administration, but daily restraint schedule decreased ethanol intake. The findings show adaptation to repeated restraint in control rats and inability of ethanol-treated rats to adapt in the stress schedule. These findings imply that excessive alcohol consumption may impair adaptation to stress and thus conceivably precipitate depression.

It is increasingly acknowledged that climate change is influencing terrestrial ecosystems by increased drought and rainfall intensities. Soil microbes are key drivers of many processes in terrestrial systems and rely on water in soil pores to fulfill their life cycles and functions. However, little is known on how drought and rainfall fluctuations, which affect the composition and structure of microbial communities, persist once original moisture conditions have been restored. Here, we study how simulated short-term drying and re-wetting events shape the community composition of soil fungi and prokaryotes. In a mesocosm experiment, soil was exposed to an extreme drought, then re-wetted to optimal moisture (50% WHC, water holding capacity) or to saturation level (100% WHC). Composition, community structure and diversity of microbes were measured by sequencing ITS and 16S rRNA gene amplicons 3 weeks after original moisture content had been restored. Drying and extreme re-wetting decreased richness of microbial communities, but not evenness. Abundance changes were observed in only 8% of prokaryote OTUs, and 25% of fungal OTUs, whereas all other OTUs did not differ between drying and re-wetting treatments. Two specific legacy response groups (LRGs) were observed for both prokaryotes and fungi. OTUs belonging to the first LRG decreased in relative abundance in soil with a history of drought, whereas OTUs that increased in soil with a history of drought formed a second LRG. These microbial responses were spread among different phyla. Drought appeared to be more important for the microbial community composition than the following extreme re-wetting. 16S profiles were correlated with both inorganic N concentration and basal respiration and ITS profiles correlated with fungal biomass. We conclude that a drying and/or an extreme re-wetting history can persist in soil microbial communities via specific response groups composed of members with broad phylogenetic origins, with possible functional consequences on soil processes and plant species. As a large fraction of OTUs responding to drying and re-wetting belonged to the rare biosphere, our results suggest that low abundant microbial species are potentially important for ecosystem responses to extreme weather events.

Fluorescence energy homotransfer offers a powerful tool for the investigation of the state of oligomerization of cell surface receptors on a cell-by-cell basis by measuring the polarized components of fluorescence intensity of cells labeled with fluorescently stained antibodies. Here we describe homotransfer-based methods for the flow cytometric detection and analysis of hetero- and homo-associations of cell surface receptors. Homotransfer efficiencies for two- and three-body energy transfer interactions are defined and their frequency distribution curves are computed from the fluorescence anisotropy distributions of multiple-labeled cells. The fractions of receptors involved in homo-clustering is calculated based on the dependence of the fluorescence anisotropy on the surface concentration of the fluorescently stained antibodies. A homotransfer analysis of the homo- and hetero-clustering of the MHCI and MHCII glycoproteins, the cytokine receptor IL-2Ralpha, transferrin receptor and the receptor-type tyrosine phosphatase CD45 on JY B and Kit-225-K6 T cells is presented. We investigated how various factors such as the type of dye, rotational mobility of the dye and dye-targeting antibody, as well as the wavelength of the exciting light affect the homotransfer. We show that the homotransfer technique combined with the high statistical resolution of flow cytometry is an effective tool for detecting different oligomeric states of receptors by using fluorophores having restricted rotational mobility on the time scale of fluorescence.

AbstractMicrobial carbon (C) use efficiency (CUE) delineates the proportion of organic C used by microorganisms for anabolism and ultimately influences the amount of C sequestered in soils. However, the key factors controlling CUE remain enigmatic, leading to considerable uncertainty in understanding soil C retention and predicting its responses to global change factors. Here, we investigate the global patterns of CUE estimate by stoichiometric modeling in surface soils of natural ecosystems, and examine its associations with temperature, precipitation, plant‐derived C and soil nutrient availability. We found that CUE is determined by the most limiting resource among these four basic environmental resources within specific climate zones (i.e., tropical, temperate, arid, and cold zones). Higher CUE is common in arid and cold zones and corresponds to limitations in temperature, water, and plant‐derived C input, while lower CUE is observed in tropical and temperate zones with widespread limitation of nutrients (e.g., nitrogen or phosphorus) in soil. The contrasting resource limitations among climate zones led to an apparent increase in CUE with increasing latitude. The resource‐specific dependence of CUE implies that soils in high latitudes with arid and cold environments may retain less organic C in the future, as warming and increased precipitation can reduce CUE. In contrast, oligotrophic soils in low latitudes may increase organic C retention, as CUE could be increased with concurrent anthropogenic nutrient inputs. The findings underscore the importance of resource limitations for CUE and suggest asymmetric responses of organic C retention in soils across latitudes to global change factors.

Abstract Tropical mountain cloud forests (TMCF) harbour a high bryophyte (mosses and liverworts) biomass and diversity. Furthermore, the high air humidity makes these forests well suited for bryophyte‐associated nitrogen (N2) fixation by cyanobacteria, providing a potentially important source of N input to the ecosystem. However, few studies have assessed bryophyte‐associated N input in these ecosystems, and these have focused on epiphytic bryophytes, whereas abundant ground‐covering bryophytes have not been included. In this study, we quantified N2 fixation rates associated with bryophytes, focusing on ground‐covering mosses in a neotropical mountain cloud forest. Furthermore, we identified the effects of climate change (higher temperature 10 vs. 20° and lower bryophyte moisture level 50% vs. 100%) on N2 fixation across bryophyte species and groups (mosses and liverworts). Nitrogen fixation rates associated with ground‐covering moss species were up to 2 kg N ha−1 year−1, which is comparable to other N inputs (e.g. N deposition) in tropical cloud forests. Furthermore, changes in temperature showed little effect on N2 fixation, but low moisture levels significantly suppressed N2 fixation activity. We found low N2 fixation activity associated with the investigated liverworts. Our results demonstrate the importance of ground‐covering, moss‐associated N2 fixation as a N source in tropical cloud forests and suggest that predicted future declines in precipitation in these systems will reduce N inputs from bryophyte‐associated cyanobacteria. Read the free Plain Language Summary for this article on the Journal blog.

Differences in bacterial community composition (BCC) between bulk and rhizosphere soil and between rhizospheres of different plant species are assumed to be strongly governed by quantitative and qualitative rhizodeposit differences. However, data on the relationship between rhizodeposit amounts and BCC are lacking. Other soil microorganisms, e.g. arbuscular mycorrhizal fungi (AMF), may also influence BCC. We simulated foliar herbivory (cutting) to reduce belowground carbon allocation and rhizodeposition of pea plants grown either with or without AMF. This reduced soil respiration, rhizosphere microbial biomass and bacteriovorous protozoan abundance, whereas none of these were affected by AMF. After labelling plants with (13)CO(2), root and rhizosphere soil (13)C enrichment of cut plants were reduced to a higher extent (24-46%) than shoot (13)C enrichment (10-24%). AMF did not affect (13)C enrichment. Despite these clear indications of reduced rhizosphere carbon-input, denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes PCR-amplified targeting DNA and RNA from rhizosphere soil did not reveal any effects of cutting on banding patterns. In contrast, AMF induced consistent differences in both DNA- and RNA-based DGGE profiles. These results show that a reduction in rhizosphere microbial activity is not necessarily accompanied by changes in BCC, whereas AMF presence inhibits proliferation of some bacterial taxa while stimulating others.