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.     

Individual PCBs as predictors for concentrations of non and mono-ortho PCBs in human milk

32 Dutch human milk samples were analyzed for PCBs with either HRGC-ECD or HRGC-LRMS in the NCI mode. Samples were collected from three different locations in The Netherlands: Amsterdam, Rotterdam and Groningen. Quantitatively, no differences could be observed between the three localities, while in addition the congener specific pattern showed a striking similarity for all individual samples. Only principal component analysis revealed slight individual differences. Based on similarities in the PCB profiles, linear relationships were calculated between 2,3′4,4′,5-PnCB (#118) or 2,2′4,4′5,5′HxCB (#153) and the most relevantnon andmonoortho PCBs exhibiting dioxinlike activity. These PCBs included 2,3,3′,4,4′-PnCB (#105), 3,3′,4,4′5-PnCB (#126) 2,3,3′,4,4′,5-HxCB (#156), 2,3,3′,4,4′,5′-HxCB (#157), 2,3′,4,4′,5,5′-HxCB (#167) and 3,3′,4,4′,5′5-HxCB (#169). Good linear relationships were observed between individual PCBs. Based on the results of this study, PCB #118 can be used to predict concentrations of the PCBs #105 and #126. PCB #153 can be used as a predictor for the PCBs #156, #157, #167 and #169, but also for the total toxic equivalencies (TEQs) ofnon andmonoortho PCBs present in human milk. This method using certain PCBs as predictors for other toxicological relevant congeners, can be useful and cost effective, e.g. for epidemiological studies. However, before applied a number of conditions should be met. These are:

1. A stable composition of the PCB matrix should be established.
2. A possible time dependent change in composition of the matrix should first be excluded when used over different time periods.

     

Toxic effects of a bloom of the diatom Rhizosolenia chunii on shellfish in Port Phillip Bay,Southeastern Australia

A bloom of the diatom Rhizosolenia chunii Karsten sensu Sundström occurred in Port Phillip Bay (38°S; 145°E), southeastern Australia, between late August and mid-October 1987. Coincident with this bloom, mussels (Mytilus edulis planulatus), scallops (Pecten alba) and flat oysters (Ostrea angasi) throughout the Bay developed a bitter taste that was so unpleasant and persistent that mussels became unmarketable for 7 mo. The bitter taste was concentrated in the digestive gland, which also showed extensive inflammation and degeneration. Digestive gland lesions were evident in mussels during September 1987, but became progressively more severe and, in the chronic end-stage were associated with high shellfish mortality 3 to 8 mo after the bloom had ceased. This study provides strong indirect evidence that the bitterness and the toxicity were caused by a bloom of R. chunii. R. chunii is the first species of diatom to be associated with shellfish mortality and, as the mortality occurred many months after the bloom had ceased, we suggest that the toxicity of certain algal blooms may be difficult to detect rather than uncommon.     

The Best-of-2-Worlds philosophy: Developing local dismantling and global infrastructure network for sustainable e-waste treatment in emerging economies

E-waste is a complex waste category containing both hazardous and valuable substances. It demands for a cost-efficient treatment system which simultaneously liberates and refines target fractions in an environmentally sound way. In most developing countries there is a lack of systems covering all steps from disposal until final processing due to limited infrastructure and access to technologies and investment. This paper introduces the ‘Best-of-2-Worlds’ philosophy (Bo2W), which provides a network and pragmatic solution for e-waste treatment in emerging economies. It seeks technical and logistic integration of ‘best’ pre-processing in developing countries to manually dismantle e-waste and ‘best’ end-processing to treat hazardous and complex fractions in international state-of-the-art end-processing facilities. A series of dismantling trials was conducted on waste desktop computers, IT equipment, large and small household appliances, in order to compare the environmental and economic performances of the Bo2W philosophy with other conventional recycling scenarios. The assessment showed that the performance of the Bo2W scenario is more eco-efficient than mechanical separation scenarios and other local treatment solutions. For equipment containing substantial hazardous substances, it demands the assistance from domestic legislation for mandatory removal and safe handling of such fractions together with proper financing to cover the costs. Experience from Bo2W pilot projects in China and India highlighted key societal factors influencing successful implementation. These include market size, informal competitors, availability of national e-waste legislation, formal take-back systems, financing and trust between industrial players. The Bo2W philosophy can serve as a pragmatic and environmentally responsible transition before establishment of end-processing facilities in developing countries is made feasible. The executive models of Bo2W should be flexibly differentiated for various countries by adjusting to local conditions related to operational scale, level of centralized operations, dismantling depth, combination with mechanical processing and optimized logistics to international end-processors.     

Derivated fetal haemoglobin as a marker for red cell age in the human fetus reflecting stimulated or impaired red blood cell production

We have determined whether derivated fetal haemoglobin (dHbF, consisting of glycated and acetylated HbF) can be used as a cell age marker for fetal red blood cells (RBCs). Cord blood was obtained between 19 and 39 weeks of gestation from 28 alloimmunised anaemic fetuses (23 RhD+ and 5 Kell) and from 20 non-anaemic fetuses and newborns (controls). Density gradient centrifugation was applied to 36 samples (20 RhD+, 15 controls and 1 Kell) to obtain fractions of increasing cell age. Blood samples were used for measurements of mean cellular volume (MCV), mean cell haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), pyruvate kinase activity (PK) and derivated fetal haemoglobin (dHbF) by cation-exchange HPLC. Reticulocytes were counted only in the whole blood samples. In all density gradient separated RBC fractions, the values for MCV, MCH and PK activity decreased and those of MCHC and dHbF increased with increasing density (equivalent to increasing cell age). The mean density was lower for RBCs of the anaemic RHD group (1.072±0.007 g/ml) than for the non-anaemic controls (1.077±0.005 g/ml) (p<0.05) The RBC density of the Kell sensitised fetus did not differ from those of the controls. In the control group, the values of the cell age markers in whole blood changed significantly with the gestational age, showing an increase of mean age of the erythrocyte population. The best linear relationship was found for dHbF (y=6.28+0.17*weeks; r=0.84; p<0.001). In the anaemic RhD+ fetuses, the RBC age markers did not change with gestational age; the dHbF percentages were lower, and the MCV, MCH, PK values and the reticulocyte counts were higher than in the controls (0.05<p<0.001). The dHbF values of the Kell sensitised fetuses were above (p<0.01) and the reticulocyte counts were below normal (p<0.05) for gestational age. For the anaemic fetuses, a significant number of the dHbF values (86%) and of the reticulocyte counts (78%) differed from the values of the controls (p<0.01). The dHbF percentages in RhD+ fetuses showed the best correlation with the Hb deficit, which is a measure for anaemia (r=−0.81, p<0.0001). We conclude that the percentage derivated HbF may indicate whether the RBC production is normal for gestational age. It may in that sense reflect stimulated or impaired erythropoiesis in alloimmunised haemolytic anaemia. Copyright © 2001 John Wiley & Sons, Ltd.     

Simultaneous estimation of jointly dependent recreation participation function

The purpose of this paper is to develop and estimate a simultaneous model of recreation participation. The logic of the model is that participation in one activity (for example, boating) is influenced by and influences participation in other activities (for example, fishing). For the empirical part of our paper we have used the simultaneous logit model developed by P. Schmidt and R. P. Strauss (Econometrics43, 745–755, 1975). For the application, we have applied the simultaneous logit model to participation in summer recreational activities in Rhode Island. Our results show that simultaneous equation models tend to forecast better, but that the specification bias from excluding the endogenous variables tends to be small.     

Water management strategies against toxic Microcystis blooms in the Dutch delta.

To prevent flooding of the Dutch delta, former estuaries have been impounded by the building of dams and sluices. Some of these water bodies, however, experience major ecological problems. One of the problem areas is the former Volkerak estuary that was turned into a freshwater lake in 1987. From the early 1990s onward, toxic Microcystis blooms dominate the phytoplankton of the lake every summer. Two management strategies have been suggested to suppress these harmful algal blooms: flushing the lake with fresh water or reintroducing saline water into the lake. This study aims at an advance assessment of these strategies through the development of a mechanistic model of the population dynamics of Microcystis. To calibrate the model, we monitored the benthic and pelagic Microcystis populations in the lake during two years. Field samples of Microcystis were incubated in the laboratory to estimate growth and mortality rates as functions of light, temperature, and salinity. Recruitment and sedimentation rates were measured in the lake, using traps, to quantify benthic-pelagic coupling of the Microcystis populations. The model predicts that flushing with fresh water will suppress Microcystis blooms when the current flushing rate is sufficiently increased. Furthermore, the inlet of saline water will suppress Microcystis blooms for salinities exceeding 14 g/L. Both management options are technically feasible. Our study illustrates that quantitative ecological knowledge can be a helpful tool guiding large-scale water management.     

Tropical agricultural residues and their potential uses in fish feeds: the Costa Rican situation

In Costa Rica as many other tropical countries, the disposal problem of agricultural wastes is widely recognized but efforts to find solutions are not equal for different sectors. This study describes the situation of major agricultural residues in Costa Rica, identifying the activities with higher amounts produced and, the potential use of these residues in fish feeds. In Costa Rica, during the 1993-1994 production season, major agricultural sectors (crop and livestock) generated a total amount of 3.15-3.25 million MT of residues (classified in by-products: used residues and wastes: not used residues). Some residues are treated to turn them into valuable items or to diminish their polluting effects (e.g., the so-called by-products). About 1.56-1.63 million MT of by-products were used for different purposes (e.g. fertilization, animal feeding, fuel, substrates in greenhouses). However, the remainder (1.59-1.62 million MT) was discharged into environment causing pollution. About 1.07-1.2 million MT wastes came from major crop systems (banana, coffee, sugarcane and oil palm) whereas the remainder came from animal production systems (porcine and poultry production, slaughtering). These data are further compared to residues estimates for the 2001-2002 production season coming from the biggest crops activities. Unfortunately, most of the studied wastes contain high levels of moisture and low levels of protein, and also contain variable amounts of antinutritional factors (e.g., polyphenols, tannins, caffeine), high fibre levels and some toxic substances and pesticides. All these reasons may limit the use of these agricultural wastes for animal feeding, especially in fish feeds. The potential use of the major vegetable and animal residues in fish feeds is discussed based on their nutritional composition, on their amount available over the year and on their pollution risks. Other constraints to use these wastes in fish feeds are the extra costs of drying and, in most cases, of transportation from several dispersed locations. It was stated that most interesting wastes are rejected green banana and coffee pulp.     

Enhancing e-waste estimates: Improving data quality by multivariate Input–Output Analysis

Waste electrical and electronic equipment (or e-waste) is one of the fastest growing waste streams, which encompasses a wide and increasing spectrum of products. Accurate estimation of e-waste generation is difficult, mainly due to lack of high quality data referred to market and socio-economic dynamics. This paper addresses how to enhance e-waste estimates by providing techniques to increase data quality. An advanced, flexible and multivariate Input–Output Analysis (IOA) method is proposed. It links all three pillars in IOA (product sales, stock and lifespan profiles) to construct mathematical relationships between various data points. By applying this method, the data consolidation steps can generate more accurate time-series datasets from available data pool. This can consequently increase the reliability of e-waste estimates compared to the approach without data processing. A case study in the Netherlands is used to apply the advanced IOA model. As a result, for the first time ever, complete datasets of all three variables for estimating all types of e-waste have been obtained. The result of this study also demonstrates significant disparity between various estimation models, arising from the use of data under different conditions. It shows the importance of applying multivariate approach and multiple sources to improve data quality for modelling, specifically using appropriate time-varying lifespan parameters. Following the case study, a roadmap with a procedural guideline is provided to enhance e-waste estimation studies.     

Monte Carlo assessment of uncertainty in the simulated hydrological response to land use change

We quantified the uncertainty in hydrological response for a set of land use change scenarios by varying plant parameters within realistic uncertainty bounds in a Monte Carlo analysis. The results show that simulated hydrological fluxes significantly change after the introduction of outwintering suckler cow management, despite the presence of a significant amount of output uncertainty due to uncertainty in the plant parameterisation. The key to a proper uncertainty assessment was to consider the uncertainty in the difference between the scenarios instead of the absolute uncertainty of each single scenario. Additionally, a sensitivity analysis showed that changing soil properties in response to land use change does not result in significantly different results in the scenario analysis.