New Silver-Gold Intensification Method of Diaminobenzidine for Double-Labeling Immunoelectron Microscopy

The available methods for double-labeling preembedding immunoelectron microscopy are highly limited because not only should the ultrastructure be preserved, but also the different antigens should be visualized by reaction end products that can be clearly distinguished in gray-scale images. In these procedures, one antigen is detected with 3,3′-diaminobenzidine (DAB) chromogen, resulting in a homogeneous deposit, whereas the other is labeled with either a gold-tagged immunoreagent, or DAB polymer, on the surface of which metallic silver is precipitated. The detection of the second antigen is usually impeded by the first, leading to false-negative results. The authors aimed to diminish this hindrance by a new silver intensification technique of DAB polymer, which converts the deposit from amorphous to granular. The method includes three major postdevelopmental steps: (1) treatment of nickel-enhanced DAB with sulfide, (2) silver deposition in the presence of hydroquinone under acidic conditions, and (3) precious metal replacement with gold thiocyanate. This new sulfide-silver-gold intensification of DAB (SSGI) allows a subsequent detection of other antigens using DAB. In conclusion, the new technique loads fine gold particles onto the DAB deposit at a very low background level, thereby allowing a reliable discernment between the elements stained for the two antigens at the ultrastructural level.     

Harvest of red-legged partridge in central Spain

A basic rule to attain sustainable use of harvested resources is to adjust take to availability. Populations of red-legged partridges in Spain have decreased in recent decades, and releases of farm-bred partridges to improve short-term availability are increasingly common. We used questionnaires and bird surveys to assess whether harvest was related to availability of wild partridges or intensity of farm-bred partridge releases. We studied 50 hunting estates, including 6 administratively labeled as intensive (with few numerical and temporal restrictions to releases). In addition, we considered hunting pressure (number of hunters × hunting days/km²) and habitat as explanatory variables in the analyses. In intensive estates, annual harvest was exclusively related to release intensity, indicating that in these estates hunting is detached from natural resources and approaches an industrial activity based on external inputs. In non-intensive estates, harvest was affected by wild stock availability, walked-up shooting pressure, and habitat (greater harvest in estates with more Mediterranean shrubland). In these estates, releases did not increase annual harvest, and can be considered an inefficient practice. Additionally, the relationship between abundance estimates and harvest disappeared in estates with low partridge abundance estimates, suggesting possibilities for overharvesting in a large proportion of estates. Increasing the abundance of wild red-legged partridge through techniques like habitat management, and improving the adjustment of harvest to availability, may be a good strategy to increase long-term harvest in non-intensive estates. Additionally, Government and managers must create ways to segregate and label the estates where only wild red-legged partridges are managed from those where releases are used, to reduce ecological costs of management decisions. © 2012 The Wildlife Society.     

Climate-based identification of suitable cropping areas for giant reed and reed canary grass on marginal land in Central and Southern Europe under climate change

Giant reed (GR) and reed canary grass (RCG) have emerged as promising perennial industrial crops for providing sustainable bioenergy from marginal land. However, there is great uncertainty among farmers and researchers about where these crops can be grown in the future due to climate change, which complicates a timely transition to a bioeconomy. Therefore, this study quantifies marginal land and suitable cropping areas for GR and RCG in Europe, as well as their overlap. To derive these areas, the present (1991–2020) and future (2071–2100, RCP8.5) growing degree days, growing season length, annual precipitation, and aridity index were analyzed using the E-OBS observational dataset and EURO-CORDEX regional climate simulations. The study concludes that while marginal land will decrease by ~18%, GR and RCG will profit from the changing European climate, increasing by ~24% and ~13%, respectively. Looking at regions of overlap between marginal land and the selected crops, a decrease of ~87% and an increase of ~462% is projected for RCG and GR, respectively. This is due to marginal land shifting southward, benefitting the warm-season grass GR, while RCG prefers cooler climates.     

Structure,productivity and energy flow in an alpine grassland in the Garhwal Himalaya

Abstract. Structure, primary production and energy flow were studied in ungrazed and grazed alpine meadows in the Garhwal Himalaya, India. In grazed grasslands, the cover of graminoids increased, the biomass of palatable species was reduced by about 50%, but the biomass of unpalatable species increased, due to reduced competition from the better-foraged species. Generally, 32% of live shoot and root biomass was consumed during a growing season. Also, above-ground plant productivity increased as a result of plant regrowth, while below-ground productivity decreased. Flow and storage of energy in ungrazed and grazed alpine grasslands showed an accumulation of 10.1 and 8.5 kJ/yr with energy conversion efficiencies of 0.7 and 0.6% respectively. Ca. 46% of the net energy accumulated was consumed by herbivores on the grazed grassland. Only 26% of the net captured energy disappeared from the system in the growing season, mainly via root and litter decomposition.     

Seed banks of grazed and ungrazed Baltic seashore meadows

Abstract. The seed banks of three grazed and three ungrazed seashore meadows were studied on the west coast of Finland. 8486 seedlings (mean 13 669 seedlings/m²) germinated from cold-treated samples (n = 343; depth = 10 cm). Most seedlings and species were monocots and perennials. The seed bank flora included 54 dicots vs. 28 monocots and 66 perennials vs. 16 annuals. The most abundant species were Juncus gerardii, Schoenoplectus tabernaemontani, Eleocharis uniglumis, Agrostis stolonifera, Juncus bufonius and Carex nigra, which made up 73% of the seed bank. Numbers of species and seedlings differed between elevation classes. Species richness was highest in elevation class 50–70 cm. The highest seed density occurred in class 20–50 cm. A model for size and species composition of the seed bank in relation to elevation is presented. The seed bank was larger and richer in species in the ungrazed than in the grazed sites, but not so in the upper elevations and closest to the open sea. Grazing reduced the size of the seed bank of Agrostis stolonifera, A. capillaris, Calamagrostis stricta, Elymus repens, Juncus bufonius, Limosella aquatica and Schoenoplectus tabernaemontani, but increased that of J. gerardii. 32 species germinated only from ungrazed samples and 11 species only from grazed ones. Multivariate classification resulted in nine sample groups. The ordination scatter was best explained by the flooding stress variables, elevation, the distance from the water line and the number of helophyte species in samples. 75 species were found both in the seed bank and in the vegetation, but there was a significant lack of resemblance (in the Mantel test) due to over-representation of some species. Eight species occurring only in the seed bank were mainly annuals or biennials (63%); those occurring only in the established vegetation (86 species) were mainly perennials (86%).