Formation of polyester blends by a recombinant strain ofPseudomonas oleovorans: Different poly(3-hydroxyalkanoates) are stored in separate granules

WhenPseudomonas oleovorans (GPo1) is grown on sodium octanoate under ammonium limiting conditions, it is able to accumulate a copolyester consisting of medium chain length 3-hydroxyalkanoic acids (PHA_m). 3-Hydroxybutyrate is only incorporated in trace amounts. WhenP. oleovorans is equipped with the PHB biosynthetic genes ofAlcaligenes eutrophus (GPo1[pVK101::PP1]), it forms a polyester containing major amounts of 3-hydroxybutyrate. The resulting polymer however is a blend of PHA_m and PHB, rather than a copolymer of 3-hydroxybutyrate and medium chain length 3-hydroxyalkanoic acids [11]. To establish whether PHA_m and PHB molecules are stored in the same or separate granules by this recombinantP. oleovorans strain, we studied polymer forming cells by freeze-fracture electron microscopy. This approach is possible because previous freeze-fracture electron microscopy studies on PHA_m and PHB accumulating strains have shown that PHA_m and PHB granules can be distinguished from each other: PHA_m granules from mushroom-like structures, whereas PHB granules from needle structures during freeze-fracturing. In this paper we show that stationary phase cells of GPo1[pVK101::PP1] contained both mushroom and needle-like structures, indicating that PHA_m and PHB chains were stored in separate granules. To be able to determine whether the separation of PHA_m and PHB is complete, the respective granules were separated on sucrose gradients. A total cell extract of GPo1[pVK101::PP1] which was subjected to sucrose gradient centrifugation revealed two white bands of different densities: the upper band with a density of 1.05 g/mL consisted exclusively of PHA_m granules, while the lower band with a density of 1.19 g/mL consisted of PHB granules only. Thus, when bacteria synthesize both PHA_m and PHB, the resulting polymer chains are segregated completely and stored in separate granules.     

Sulfide assimilation by ectosymbionts of the sessile ciliate, <Emphasis Type="Italic">Zoothamnium niveum</Emphasis>

We investigated the constraints on sulfide uptake by bacterial ectosymbionts on the marine peritrich ciliate Zoothamnium niveum by a combination of experimental and numerical methods. Protists with symbionts were collected on large blocks of mangrove-peat. The blocks were placed in a flow cell with flow adjusted to in situ velocity. The water motion around the colonies was then characterized by particle tracking velocimetry. This shows that the feather-shaped colony of Z. niveum generates a unidirectional flow of seawater through the colony with no recirculation. The source of the feeding current was the free-flowing water although the size of the colonies suggests that they live partly submerged in the diffusive boundary layer. We showed that the filtered volume allows Z. niveum to assimilate sufficient sulfide to sustain the symbiosis at a few micromoles per liter in ambient concentration. Numerical modeling shows that sulfide oxidizing bacteria on the surfaces of Z. niveum can sustain 100-times higher sulfide uptake than bacteria on flat surfaces, such as microbial mats. The study demonstrates that the filter feeding zooids of Z. niveum are preadapted to be prime habitats for sulfide oxidizing bacteria due to Z. niveum’s habitat preference and due to the feeding current. Z. niveum is capable of exploiting low concentrations of sulfide in near norm-oxic seawater. This links its otherwise dissimilar habitats and makes it functionally similar to invertebrates with thiotrophic symbionts in filtering organs. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Parameterizing,evaluating and comparing metapopulation models with data from individual-based simulations

Due to the lack of sufficient data and appropriate ecological information parameterizing predictive population dynamical models usually is a difficult task. The approach proposed in this study is meant to overcome this problem by using detailed individual-based simulations to generate artificial data. With short-term data samples, the models to be investigated can be parameterized and their predictions be compared. The flexibility of individual-based simulations as experimental tools also facilitates the evaluation and comparison of different (aggregated) model types. The presented approach is a step towards unifying models of different complexity. As an example we applied it to two metapopulation models of insect species in a highly fragmented landscape: the well-known incidence function model with a patch-based representation of space and a grid-based analogue. The models are tested with respect to their data requirement and recommendations for a better data sampling are derived.     

Plant surface wax affects parasitoid’s response to host footprints

The plant surface is the substrate upon which herbivorous insects and natural enemies meet and thus represents the stage for interactions between the three trophic levels. Plant surfaces are covered by an epicuticular wax layer which is highly variable depending on species, cultivar or plant part. Differences in wax chemistry may modulate ecological interactions. We explored whether caterpillars of Spodoptera frugiperda, when walking over a plant surface, leave a chemical trail (kairomones) that can be detected by the parasitoid Cotesia marginiventris. Chemistry and micromorphology of cuticular waxes of two barley eceriferum wax mutants (cer-za.126, cer-yp.949) and wild-type cv. Bonus (wt) were assessed. The plants were then used to investigate potential surface effects on the detectability of caterpillar kairomones. Here we provide evidence that C. marginiventris responds to chemical footprints of its host. Parasitoids were able to detect the kairomone on wild-type plants and on both cer mutants but the response to cer-yp.949 (reduced wax, high aldehyde fraction) was less pronounced. Experiments with caterpillar-treated wt and mutant leaves offered simultaneously, confirmed this observation: no difference in wasp response was found when wt was tested against cer-za.126 (reduced wax, wt-like chemical composition) but wt was significantly more attractive than cer-yp.949. This demonstrates for the first time that the wax layer can modulate the detectability of host kairomones.     

The safety barometer: How safe is my plant today? Is instantaneously measuring safety level utopia or realizable?

During the last decade, serious accidents have continued to occur in the process industry. Apparently the scenarios of various undesired events leading to those accidents are still not sufficiently controlled. The key question is how potentially hazardous situations develop, what processes form the basis for this development, and how to control them? Safety level is not static but depends on many risk factors that change in presence and intensity over location and time. Safety level is dependent not only on technical process parameters that have immediate effects on the ‘frequency’ or probability of catastrophic consequences, but also depends on equipment integrity degradation, operational and management quality, attitudes, and cultural processes which may change over a prolonged time. The time and human interaction aspects make dynamic risk assessment complex. This paper will outline a conceptual approach using in addition to the regular process parameter signals received, also weak and slowly changing signals from various safety indicators, enabling to keep track of the risk factors. In theory this could lead to obtaining an instantaneous safety level ‘measure’ making possible forecast alarming for an imminent event to occur. Such concept could be regarded as a ‘writing’ safety barometer, or barograph. However, there are quite a number of problems to be solved which in the paper will be discussed.     

A mathematical model for the distribution and abundance of benthic algae in a Norwegian fjord

A mathematical model has been developed to study the distribution and abundance of benthic algae species in a Norwegian fjord. The main forcing functions are the physical factors which have well defined gradients in this area, but biotic interactions are also accounted for in the model.The distribution of algae along vertical transects in the intermediate fjord area has been simulated, and the observed distribution is reproduced fairly well. This indicates that the major factors determining the equilibrium elevation between intertidal and subtidal vegetations in this area are adequately represented in the model.Based on observations in a fjord branch with a high level of freshwater influx and polluted by heavy metals, assumptions are made of the combined influence of these contaminants on growth characteristics of the species considered. The resulting effects on community development and community patterns are simulated. For a certain level of contamination, the decrease in abundance, community development rate and species separation have been determined. It is also shown that the fluctuations in biomass during a year are more pronounced than under unaffected conditions.The relative importance of external growth regulating factors and self-shading is demonstrated for different degrees of community development, and we have indicated that this may be an important factor in the assessment of transient and long-term responses to “natural” and man-made influences on the shore-line vegetation.One of the main problems during model development was the formulation of the interaction function for factors influencing growth parameters. Several alternatives were examined.The differences between the most frequently used forms were small with respect to abundance and not very large with respect to possible changes in distribution. It was noted that even by including only the most limiting factor at each instant, (Liebig's form) a high percentage of the potentially limiting factors did actually limit growth during periods of the year.The model is intended as a working tool for the studies of changes in the shore-line vegetation caused by man-made interferences like heavy metal and oil pollution, and we believe that the results presented demonstrate that it can successfully be used for this purpose.