Characterization of Oat Endoproteinases that Hydrolyze Oat Globulins

During the germination of oats, the major seed storage proteins (globulins) are hydrolyzed by endoproteinases. We have used two methods to characterize these endoproteinases. A qualitative PAGE method that used oat globulins as gel‐incorporated substrates was used to determine which enzymes hydrolyzed the globulins. The proteolytic hydrolysis products were studied by hydrolyzing the globulins in vitro with the endoproteinases and analyzing the products by SDS‐PAGE. Class‐specific proteinase inhibitors were used to show that the globulin hydrolyzing enzymes were cysteine‐class proteinases. The proteinases were active at pH 3.8. Using the gel analysis method, a little activity was present at the beginning of seed germination, but the major activity only appeared on the sixth day of germination. Extracts from four‐day germinated oats contained cysteine proteinases that hydrolyzed the globulins in vitro to form a polypeptide of intermediate size (MW ≈34,500). Cysteine proteases from an eight‐day germinated sample totally hydrolyzed the globulins in <1 hr. Very little hydrolysis occurred at pH 6.2, the pH of germinated oats endosperm tissue. The fact that hydrolysis occurred quickly at pH 3.8 implies that there is probably pH compartmentalization within the endosperm, with some areas of the seed having a low pH value where the globulins can be degraded.     

The microbiota of pigs influenced by diet texture and severity of Lawsonia intracellularis infection

Pigs with and without naturally occurring Lawsonia intracellularis infection were fed diets with different texture. In a previous study from 79 pig herds using a similar feeding on pelleted or non-pelleted form showed that the non-pelleted diet was associated with a reduced prevalence of L. intracellularis. In this study a mechanistic approach was taken for explaining and testing this observation by studying the microbiota and the occurrence of L. intracellularis in the distal ileum of 54 pigs by terminal restriction fragment length polymorphism (T-RFLP) analysis, Real-Time PCR and in situ hybridization. The texture of the diet influenced the microbiota, and from a quantitative discriminative analysis of the terminal restriction fragments (T-RFs) of ileum samples it was deduced that Clostridium spp. and Lactobacillus spp. were associated with the non-pelleted diet and Streptococcus spp. with the pelleted diet. In experimentally infected pigs it was verified that 89bp and 90bp sized T-RFs (HhaI) from ileum represented L. intracellularis. The non-pelleted diet seemed to reduce the relative amount of L. intracellularis in the total microbiota of the ileum, but the number of pigs detected positive with L. intracellularis by Real-Time PCR was not influenced. The five pigs with highest L. intracellularis content showed T-RFs that were not present in profiles from less or non-infected pigs, which may indicate that some bacterial species were associated with L. intracellularis infection.     

Neither hippurate-negative Brachyspira pilosicoli nor Brachyspira pilosicoli type strain caused diarrhoea in early-weaned pigs by experimental infection

A hippurate-negative biovariant of Brachyspira pilosicoli (B. pilosicolihipp-) is occasionally isolated in diarrhoeic pigs in Finland, often concomitantly with hippurate-positive B. pilosicoli or Lawsonia intracellularis. We studied pathogenicity of B. pilosicolihipp- with special attention paid to avoiding co-infection with other enteric pathogens. Pigs were weaned and moved to barrier facilities at the age of 11 days. At 46 days, 8 pigs were inoculated with B. pilosicolihipp- strain Br1622, 8 pigs were inoculated with B. pilosicoli type strain P43/6/78 and 7 pigs were sham-inoculated. No signs of spirochaetal diarrhoea were detected; only one pig, inoculated with P43/6/78, had soft faeces from day 9 to 10 post inoculation. The pigs were necropsied between days 7 and 23 after inoculation. Live pigs were culture-negative for Brachyspira spp., but B. pilosicolihipp- was reisolated from necropsy samples of two pigs. The lesions on large colons were minor and did not significantly differ between the three trial groups. In silver-stained sections, invasive spirochaetes were detected in colonic mucosae of several pigs in all groups. Fluorescent in situ hybridisation for genus Brachyspira, B. pilosicoli and strain Br1622 was negative. However, in situ detection for members of the genus Leptospira was positive for spirochaete-like bacteria in the colonic epithelium of several pigs in both infected groups as well as in the control group. L. intracellularis, Salmonella spp., Yersinia spp. and intestinal parasites were not detected. The failure of B. pilosicoli strains to cause diarrhoea is discussed with respect to infectivity of the challenge strains, absence of certain intestinal pathogens and feed and management factors.     

Flood or drought: how do aerosols affect precipitation?

Aerosols serve as cloud condensation nuclei (CCN) and thus have a substantial effect on cloud properties and the initiation of precipitation. Large concentrations of human-made aerosols have been reported to both decrease and increase rainfall as a result of their radiative and CCN activities. At one extreme, pristine tropical clouds with low CCN concentrations rain out too quickly to mature into long-lived clouds. On the other hand, heavily polluted clouds evaporate much of their water before precipitation can occur, if they can form at all given the reduced surface heating resulting from the aerosol haze layer. We propose a conceptual model that explains this apparent dichotomy.     

Degradation of Secalins During Rye Sourdough Fermentation

Rye sourdough (RSD) gives rye bread mildly acidic taste and desired flavor. Flavor precursors (amino acids and small peptides) are generated in the proteolytic breakdown of rye proteins. Our aim was to study the protein degradation during RSD fermentations. Two sourdoughs were prepared of flours derived from two rye cultivars (Amilo and Akusti). RSD samples were collected during fermentations. Three protein fractions were obtained by sequential protein extraction and these were analyzed by SDS‐PAGE. Free amino nitrogen (FAN) was measured with a ninhydrin method. In addition, two rye incubations without starter microorganisms (with antibiotics) were made at pH 3.6 and 6.1, and proteinase profiles of the rye cultivars were analyzed at pH 4.3. SDS‐PAGE analysis showed that during RSD fermentations, rye proteins, especially the alcohol‐soluble secalins, were degraded. Secalins also evidently degraded during the incubation without starter microorganisms at pH 3.6. Aspartic proteinases were in the major proteinase group in both rye cultivars. This study confirms that endogenous proteinases of rye, mainly aspartic proteinases, hydrolyze rye proteins, especially secalins, during RSD fermentation. Protein degradation in rye sourdoughs may thus be enhanced by selecting rye flours with high proteolytic activity toward secalins.     

Differentiating sources of CO2 from organic soil under bioenergy crop cultivation: A field-based approach using C

We tested here a plant-soil system to separate recent, plant-derived and native, soil-derived carbon in soil respiration. The approach uses a perennial crop cultivated on an organic soil where upper soil layers have been removed as a result of peat extraction. There, the ¹⁴C signal from native organic matter is highly depleted compared to that in vegetation established at the site after peat extraction ceased. Radiocarbon was analyzed in carbon dioxide respired from soil over one growing season, and a two-pool isotope mixing model was applied to calculate the relative contribution of old vs. new carbon sources. The analysis showed that the approach is reliable for source partitioning with isotopes. After six years of cultivation, old peat decomposition contributed less to total soil respiration than respiration of recent plant material (30% vs. 70% on average, respectively), but the relative proportions were highly variable over the growing season. The approach offers a new possibility to follow the fate of old, native soil organic matter in highly organic soils.     

Aerosol particles in the developing world; a comparison between New Delhi in India and Beijing in China

In developing countries, aerosol particles damage the health of hundreds of millions of people. Migration from the country side to megacities increases emissions and exposure to particles. Some countries have started to limit emissions based on particulate mass, but this may increase particle number concentrations. In this study we discuss some earlier measurements carried out in the developing world and compare results from one-week measurement campaigns concerning the particle number size distribution and PM₁₀ mass concentrations in New Delhi, India and Beijing, China. Our results show that submicron particle concentrations are high in both places. The average PM₁₀ concentration was 360 μg/m³ in New Delhi and 120 μg/m³ in Beijing. The corresponding total particle number concentrations in the size range 3–800 nm were 63 000 cm^?3 and 35 000 cm^?3. Number and mass concentrations and their characteristics showed significantly different behaviour between these two locations, which stresses the importance of long-term simultaneous measurements of both quantities in different types of megacities.     

Dispersal,environment, and floristic variation of western Amazonian forests

The distribution of plant species, the species compositions of different sites, and the factors that affect them in tropical rain forests are not well understood. The main hypotheses are that species composition is either (i) uniform over large areas, (ii) random but spatially autocorrelated because of dispersal limitation, or (iii) patchy and environmentally determined. Here we test these hypotheses, using a large data set from western Amazonia. The uniformity hypothesis gains no support, but the other hypotheses do. Environmental determinism explains a larger proportion of the variation in floristic differences between sites than does dispersal limitation; together, these processes explain 70 to 75% of the variation. Consequently, it is important that management planning for conservation and resource use take into account both habitat heterogeneity and biogeographic differences.     

Marine connectivity in spatial conservation planning: analogues from the terrestrial realm

Context

Spatial prioritization is an analytical approach that can be used to provide decision support in spatial conservation planning (SCP), and in tasks such as conservation area network design, zoning, planning for impact avoidance or targeting of habitat management or restoration.

Methods

Based on literature, we summarize the role of connectivity as one component of relevance in the broad structure of spatial prioritization in both marine and terrestrial realms.

Results

Partially diffuse, directed connectivity can be approximated in Zonation-based multi-criteria SCP by applying hydrodynamic modelling, knowledge on species traits, and information on species occurrences and quality of habitats. Sources and destinations of larvae or propagules can be identified as separate spatial layers and taken into account in full-scale spatial prioritization involving data on biota, as well as economic factors, threats, and administrative constraints. While population connectivity is an important determinant of metapopulation persistence, the importance of marine connectivity depends on species traits and the marine environment studied. At one end of the continuum are species that occupy isolated habitats and have long pelagic larval durations in deeper sea areas with strong directional currents. At the other extreme are species with short pelagic durations that occupy fragmented habitats in shallow topographically complex sea areas with weak and variable currents.

Conclusions

We conclude that the same objectives, methods, and analysis structures are applicable to both terrestrial and marine spatial prioritization. Marine spatial conservation planning, marine spatial planning, marine zoning, etc., can be implemented using methods originated in the terrestrial realm of planning.