A.C. van Aelst

Cluster Structure of Anaerobic Aggregates of an Expanded Granular Sludge Bed Reactor

ABSTRACT The metabolic properties and ultrastructure of mesophilic aggregates from a full-scale expanded granular sludge bed reactor treating brewery wastewater are described. The aggregates had a very high methanogenic activity on acetate (17.19 mmol of CH4/g of volatile suspended solids [VSS]·day or 1.1 g of CH4 chemical oxygen demand/g of VSS·day). Fluorescent in situ hybridization using 16S rRNA probes of crushed granules showed that 70 and 30% of the cells belonged to the archaebacterial and eubacterial domains, respectively. The spherical aggregates were black but contained numerous whitish spots on their surfaces. Cross-sectioning these aggregates revealed that the white spots appeared to be white clusters embedded in a black matrix. The white clusters were found to develop simultaneously with the increase in diameter. Energy-dispersed X-ray analysis and back-scattered electron microscopy showed that the whitish clusters contained mainly organic matter and no inorganic calcium precipitates. The white clusters had a higher density than the black matrix, as evidenced by the denser cell arrangement observed by high-magnification electron microscopy and the significantly higher effective diffusion coefficient determined by nuclear magnetic resonance imaging. High-magnification electron microscopy indicated a segregation of acetate-utilizing methanogens (Methanosaeta spp.) in the white clusters from syntrophic species and hydrogenotrophic methanogens (Methanobacterium-like andMethanospirillum-like organisms) in the black matrix. A number of physical and microbial ecology reasons for the observed structure are proposed, including the advantage of segregation for high-rate degradation of syntrophic substrates.

Endosperm cap weakening and endo-beta-mannanase activity during priming of tomato (Lycopersicon esculentum cv. Moneymaker) seeds are initiated upon crossing a threshold water potential.

The relationship between endosperm cap weakening and endo-β-mannanase activity during priming and the time to germination after priming was studied in tomato ( Lycopersicon esculentum ) seeds. During priming of seeds in −0.4 MPa PEG, the mechanical restraint of the endosperm cap decreased while the endo-β-mannanase activity in the endosperm cap increased. There was no decrease in required puncture force and no increase in endo-β-mannanase activity in seeds during priming in −1.0 MPa PEG. Two classes of seeds could be distinguished during priming in −0.7 MPa PEG: one with decreased required puncture force and one without. A strong correlation was found between the lowering of the mechanical restraint and endo-β-mannanase activity. It was concluded that individual seeds have to cross a threshold water potential in order to develop enzyme activity and lower their mechanical restraint. A decrease in required puncture force and increase in endo-β-mannanase activity correlated with ice crystal-induced porosity in the endosperm cap cell walls in scanning micrographs. It was presumed that ice crystal-induced porosity reflects cell-wall hydrolysis. Germination time after priming correlated positively with required puncture force during priming, depending on the osmotic potential. Seeds in −1.0 MPa PEG improved their time to germination, without a decrease in the required puncture force. Therefore it was concluded that lowering of the endosperm restraint during priming positively affects the time to germination of primed seeds but is not a prerequisite for rapid germination.

Is oxidative stress involved in the loss of neem (Azadirachta indica) seed viability

Neem (Azadirachta indica) is a valuable multipurpose tree of tropical arid and semi-arid regions. The use of its seeds is hindered by their short storage longevity. The possible causes of rapid loss of viability were investigated on different seed lots during exposure to 32% and 75% RH at 20°C. Within 6 months the seeds almost lost germinability at 75% RH, whereas at 32% RH viability decreased only slightly. On rehydration, the axis cells from nongerminable seeds had lost turgor, whereas those from viable seeds were turgescent as visualized by low temperature scanning electron microscopy images of fractured axes. Glutathione oxidation status was used to estimate oxidative stress during storage. Oxidative stress was much higher at 75% RH storage than at 32% RH, mainly caused by the rapid loss of reduced glutathione at 75% RH. Oligosaccharides and phospholipids decreased, and free fatty acids increased during storage at the high RH but remained at a constant level at the low RH. However, the degree of fatty acid unsaturation between viable and nonviable seed lots was similar. During the (slow) dehydration of fresh seeds, total glutathione, oligosaccharides and phospholipids accumulated, particularly in the initially more hydrated seeds. We interpret this accumulation as a post-maturation process associated with acquisition of the capability for long-term survival in the dry state. The mass ratio of oligosaccharides to sucrose was 0.19 on average in dehydrated neem seeds. The data suggest that the storage behaviour of neem seeds has features that characterize it as orthodox.

Combined chemical and electrical enhancement modulates stratum corneum structure

In a previous in vitro study it has been shown that pretreatment with a water-based surfactant formulation results in a two-fold increase in transdermal iontophoretic transport of R-apomorphine compared to iontophoresis only. The aim of the study presented in this paper was to unravel the mechanisms involved in the increased iontophoretic delivery. Freeze fracture electron microscopy and cryo-scanning electron microscopy were used to visualise the ultrastucture of human stratum corneum after (i) application of the surfactant formulation, (ii) iontophoresis and (iii) application of the surfactant formulation followed by iontophoresis. Non-occlusive application of the surfactant formulation did not exert any detectable changes in the ultrastructure of the stratum corneum, except for swelling of the outermost corneocyte layers. Application of a current density of 0.5 mA/cm(2) for 9 h induced a swelling of the corneocytes and the formation of water pools that were occasionally present in the intercellular regions. Application of the surfactant formulation followed by iontophoresis resulted in a further swelling of the corneocytes and a frequent presence of water pools in the intercellular regions throughout the whole stratum corneum. The observed changes in the ultrastructure of the stratum corneum can explain the increased R-apomorphine transport during iontophoresis.

Nuclear pore dynamics during pollen development and androgenesis in Brassica napus.

Abstract Changes in nuclear pore complex (NPC) densities, NPCs/nucleus and NPCs/µm3, are described using freeze-fractured Brassica napus microspores and pollen in vivo and in vitro. Early stages of microspore- and pollen-derived embryogenic cells were also analysed. The results of in vivo and in vitro pollen development indicate an increase in activity of the vegetative nucleus during maturation of the pollen. At the onset of microspore and pollen culture, NPC density decreased from 15 NPCs/µm2 at the stage of isolation to 9 NPCs/µm2, under both embryogenic and non-embryogenic conditions. This implies that the drop in NPC density might be a result of culturing the microspores and pollen rather than an indication for microspore and pollen embryogenesis in Brassica napus. However, after 1 day in culture under embryogenic conditions, the NPC density increased again and stabilised around 13 NPCs/µm2, whereas under non-embryogenic conditions the NPC density remained about 9 NPCs/µm2. This low density of 9 NPCs/µm2 was also found in the nuclei of sperm cells, in contrast to the 19 NPCs/µm2 found in the vegetative nucleus. It means that, although both the vegetative and sperm nuclei are believed to be metabolically rather inactive in mature pollen, the NPC density of vegetative nucleus is twice as high as the NPC density of the sperm nuclei. In a few cases, embryos formed suspensor-like structures with a NPC density of 9 NPCs/µm2, indicating a lower nucleocytoplasmic exchange of the nuclei of the suspensor cells than with the nuclei in the embryo proper. In addition, observations on NPCs and other organelles, obtained by high resolution cryo-scanning microscopy, are presented.

Biological Production of Selenium Nanoparticles from Waste Waters

Due to the photoelectric, semi-conducting and X-ray-sensing properties, selenium is an element of great economic potential. Since nanomaterials display special properties compared to bulk particles, the production of selenium nanostructures (wires, rods or spherical particles) has attracted much attention. In this study, alternative approaches to classical synthesis were investigated, producing selenium nanospheres biologically during treatment of contaminated water.