Who's in and why? A typology of stakeholder analysis methods for natural resource management

Stakeholder analysis means many things to different people. Various methods and approaches have been developed in different fields for different purposes, leading to confusion over the concept and practice of stakeholder analysis. This paper asks how and why stakeholder analysis should be conducted for participatory natural resource management research. This is achieved by reviewing the development of stakeholder analysis in business management, development and natural resource management. The normative and instrumental theoretical basis for stakeholder analysis is discussed, and a stakeholder analysis typology is proposed. This consists of methods for: i) identifying stakeholders; ii) differentiating between and categorising stakeholders; and iii) investigating relationships between stakeholders. The range of methods that can be used to carry out each type of analysis is reviewed. These methods and approaches are then illustrated through a series of case studies funded through the Rural Economy and Land Use (RELU) programme. These case studies show the wide range of participatory and non-participatory methods that can be used, and discuss some of the challenges and limitations of existing methods for stakeholder analysis. The case studies also propose new tools and combinations of methods that can more effectively identify and categorise stakeholders and help understand their inter-relationships.     

Integral assessment of estrogenic potentials in sediment-associated samples

Background, aim, and scope  The enzyme-linked receptor assay (ELRA) detects estrogenic and anti-estrogenic effects at the molecular level of receptor binding and is a useful tool for the integrative assessment of ecotoxicological potentials caused by hormonally active agents (HAA) and endocrine disrupting compounds (EDC). The main advantage of the ELRA is its high sample throughput and its robustness against cytotoxicity and microbial contamination. After a methodological adaptation to salinity of the ELRA, according to the first part of this study, which increased its salinity tolerance and sensitivity for 17-β-estradiol, the optimised ELRA was used to investigate 13 native sediments characterised by different levels of salinity and chemical contamination. The applicability of the ELRA for routine analysis in environmental assessment was evaluated. Salinity is often a critical factor for bioassays in ecotoxicological sediment assessment. Therefore, salinity of the samples was additionally adjusted to different levels to characterise its influence on elution and binding processes of receptor-binding substances. Materials and methods  The ELRA was carried out with the human estrogen receptor α (ER) in a 96-well microplate format using the experimental setup known from the competitive immunoassay based on ligand–protein interaction. It is an important improvement that a physiologically relevant receptor was used as a linking protein instead of an antibody. The microplates were coated with a 17-β-estradiol-BSA conjugate, and dilution series of estradiol and of native sediment samples were added and incubated with the ER. After a washing step, a biotinylated mouse anti-ER antibody was added to each well. Receptor binding to estradiol, agonistic and antagonistic receptor binding, were determined by a streptavidin-POD-biotin complex with subsequent measurement of the peroxidase activity at the wavelength of 450 nm using a commercial ELISA multiplate reader. The sediment elutriates and pore water samples of sediments were tested in a dilution series to evaluate at which dilution step the receptor-binding potential ends. In the elution process (see Section 2.1 to 2.2), a method was developed to adjust the salinity to the levels of the reference testings, which offers an appropriate option to adjust the salinity in both directions. Statistical evaluation was made with a combination of the Mann–Whitney U test and the pT-method. Results  This part of the study characterised the environmental factor ‘salinity’ for prospective applications of the ELRA. Using reference substances such as 17-β-estradiol, the ELRA showed sigmoid concentration-effect relations over a broad range from 0.05 μg/l to 100 μg/l under physiological conditions. After methodological optimisation, both sensitivity and tolerance of the assay against salinity could be significantly raised, and the ELRA became applicable under salinity conditions up to concentrations of 20.5‰. The mean relative inter-test error (n = 3) was around 11% with reference substances and below 5% for single sediments elutriates in three replicates each. For sediment testings, the pore water and different salinity-adjusted elutriates of 13 sediments were used. A clear differentiation of the receptor-binding potential could be reached by application of the pT-method. Thereby, pT-values from one to six could be assigned to the sediments, and the deviation caused by the different salinity conditions was one pT-value. The mean standard deviation in the salinity adaptation procedure of the elutriates was below 5%. Discussion  Although the ELRA has already been used for assessments of wastewater, sludge and soil, its applicability for samples to different salinity levels has not been investigated so far. Even if the ELRA is not as sensitive as the E-screen or the YES-assay, with regard to reference substances like 17-β-estradiol, it is a very useful tool for pre-screening, because it is able to integrate both estrogenic as well as anti-estrogenic receptor-binding effects. According to the results of sediment testing, and given the integrative power to detect different directions of effects, the ELRA shows sufficient sensitivity and salinity tolerance to discriminate receptor-binding potentials in environmental samples. Conclusions  The optimised ELRA assay is a fast, cost-effective, reliable and highly reproducible tool that can be used for high-throughput screening in a microplate format in detecting both estrogenic and anti-estrogenic effects. Additionally, the ELRA is robust against microbial contaminations, and is not susceptible towards cytotoxic interferences like the common cell-culture methods. The general applicability and sufficient sensitivity of the ELRA was shown in freshwater environments. Marine and brackish samples can be measured up to salinity levels of 20.5‰. Recommendations and perspectives  In view of the proven sensitivity, functionality and the fastness of the ELRA, it is recommendable to standardise the test method. At the moment, no adequate in vitro test procedure exists which is standardised to DIN or ISO levels. The E-screen and the yeast estrogen/androgen screens (YES/YAS) sometimes underlie strong cytotoxic effects, as reported in the first part of this study. Further development of an ELRA assay using human androgen receptors appears to be very promising to gain information about androgenic and anti-androgenic effects, too. This would offer a possibility to use the ELRA as a fast and reliable pre-screening tool for the detection of endocrine potentials, thus minimising time and cost-expensive animal experiments.     

Investigating Jacaranda mimosifolia tree as biomonitor of atmospheric trace metals

Studies on the use of tree bark as biomonitors for environmental pollutants are still very scarce. We evaluated the reliability of using Jacaranda mimosifolia, a common tree in Tshwane City of South Africa, as a suitable biomonitor of atmospheric trace metals. Bark samples were collected from ten different locations during two sampling periods. The concentrations of the metals were determined by inductively coupled plasma mass spectrometry. The concentrations of the metals were 33.2–1,795 μg/g (Pb), 21.4–210 μg/g (Cu), 68.4–490 μg/g (Zn), 30.6–2,916 μg/g (Cr), 0.12–1.34 μg/g (Cd), and 6.04–68.0 μg/g (V), respectively. The differences obtained for the results from different sites were significant (p?< 0.05). A significant difference was also observed between the two sampling periods. The trace metals concentrations suggested that automobile emissions are a major source of these metals. The study also confirms the suitability of J. mimosifolia as a biomonitor of atmospheric deposition of these metals.     

Anaerobic inhibition and biodegradation of antibiotics in ISO test schemes

Municipal sewage is the main exposure route for antibiotics that are used in human medical care. Antibiotics that adsorb to the primary sludge and/or sur-plus activated sludge will enter the anaerobic digesters of municipal sewage treatment plants. Here anaerobic biodegradation or inhibition of anaerobic bacteria resulting in a disturbance of the process might occur. ISO standards 13641 (2003) and 11734 (1999) were used for assessing the anaerobic inhibition of 16 and the anaerobic biodegradability of 9 antibiotics respectively. Digestion sludge from a municipal sewage treatment plant (1g/l d.s.) was used as inoculum in both tests. In ISO 13641 (2003) most antibiotics showed only moderate inhibition effects after a 7 day incubation period, with EC50 values between 24 mg/l and more than 1000 mg/l (equal to mg/g d.s.). In contrast, metronidazol was decisively toxic to anaerobic bacteria with an EC50 of 0.7 mg/l. In the anaerobic degradation tests according to ISO standard 11734 (1995), only benzylpenicillin showed certain ultimate biodegradation after 60 days and most antibiotics inhibited the digesting sludge in the respective parallel tested inhibition controls. Thus the inhibition of anaerobic bacteria by antibiotics observed in the degradation tests was higher than expected from the results of the inhibition tests. The possible explanations are that distinct substrates are used (yeast extract versus sodium benzoate), that the digestion sludge loses activity during the washing steps performed for the degradation tests and that the exposure time in the degradation tests was 8 times longer than in the inhibition test.     

Vertical profile of PCDD/Fs, dioxin-like PCBs, other PCBs, PAHs, chlorobenzenes, DDX, HCHs, organotin compounds and chlorinated ethers in dated sediment/soil cores from flood-plains of the river Elbe, Germany

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) and other organic micropollutants were determined in dated sediment/soil cores collected from the flood-plain of the river Elbe near Pevestorf (PT), approximately 125 km upstream of Hamburg, and Heuckenlock (HL) in southeast of Hamburg. Concentrations of PCDD/Fs peaked sharply at PT in the 1950s and at HL at the end of the 1940s. Cluster analyses provide evidence that the region of Bitterfeld-Wolfen (about 350-400 km upstream of Hamburg) could be the source of the PCDD/F contamination existing in the cores PT and HL since the 1940s. Obviously it is caused by sediments of the river Elbe of a similar composition. Whereas the PCDD/Fs, HCHs (hexacyclohexane isomers), DDX (DDT, DDD, DDE), and tetrachlorinated ethers in PT and HL presumably originated predominantly from the Bitterfeld-Wolfen region, organotin compounds in HL and dichlorinated haloethers in HL during the 1940s and 1950s can probably largely be attributed to emissions from the Hamburg region. Although they are separated by a large distance, in both sediment cores PT and HL concentrations and composition patterns of most organic micropollutants analyzed widely match. Inductively it can be concluded that similar contaminations will be found in many of the river bank soils between the Bitterfeld-Wolfen region and Hamburg. Excavation of top soils may uncover highly contaminated materials. Since the dated sediment cores show the variation in contaminants in the Elbe sediments over a defined time period, it is possible to make an approximate assessment of the actual degree of contamination to be expected in areas where in previous decades contaminated dredged sediments from the Elbe and from the Port of Hamburg have been deposited on land and used for building plots or for agricultural purposes.     

Sustainability, substance flow management and time. Part I Temporal analysis of substance flows

Flows of chemical substances need to be managed in a sustainable way. Sustainable development as a whole and the sustainable management of substance flows in particular are both time issues. These include the importance of the dynamics of substance flows and the way these interconnect with the use of resources, the avoidance of environmental pollution, and their effects on health and food production. Another prerequisite for the proper management of substance flows is justice within and between generations. This requires a systematic approach and a systematic analysis of the issues as well as of the actions to be taken. One tool for such a systematic approach is temporal analysis. It brings the temporal aspects of the substances themselves and of their intended use, as well as factors affecting the stakeholders, such as decision makers, producers and consumers, into focus. In the past, timing factors were rarely taken into account. Knowledge of the temporal dynamics of substance flows and their resultant outcomes, as well as of their interaction with ecological, economic and social systems, is a basic requirement for successful substance flow management. The need to include temporal aspects into substance flow management and how to do so is outlined here. Included are not only politicians but also practitioners and scientists who must explicitly take into account adequate time scales, points in time, breaks and other forms of time in planning and acting.     

Anti-sense expression of putrescine N-methyltransferase confirms defensive role of nicotine in Nicotiana sylvestris against Manduca sexta

Summary. Several lines of evidence support the defensive function of nicotine production in the Nicotiana genus against a range of herbivores, but the evidence is largely correlative. To suppress nicotine production in planta and to test its defensive function, we expressed DNA of putrescine N-methyl transferase in an anti-sense orientation (AS-PMT) in N. sylvestris and fed leaf material from two lines of transformed and wild type plants to Manduca sexta larvae. Larvae consumed more leaf area and gained more mass on the foliage of plants with low PMT expression and low nicotine levels as compared to plants with high PMT expression and high nicotine levels and wild type plants. Overall, larval consumption and performance were negatively correlated with constitutive nicotine levels. We conclude that nicotine decreases the palatability of N. sylvestris leaves to the nicotine-resistant M. sexta larvae.     

Attribution of Carbon Dioxide Fluxes to Crop Types in a Heterogeneous Agricultural Landscape of Argentina

The increasing proportion of agricultural lands worldwide makes it necessary to intensify the research concerning the carbon exchange at agricultural sites. In order to determine the Net Ecosystem Exchange (NEE) in an agricultural landscape in the province of Buenos Aires, Argentina, we carried out eddy covariance measurements with a flux tower, which was placed between two agricultural fields. Therefore, the measured CO₂ flux represents the accumulated flux from both areas, i.e., from different crop types. We here present an analysis method which attributes the flux to the two crop types. For this analysis, we applied the Hsieh footprint model to identify the contributing source area to the flux measurement. We then applied a multiple regression analysis to calculate the NEE in the growing season 2011/2012 for each field separately. The pronounced differences in the time courses of the CO₂ fluxes in the two fields can be explained by the different sowing times and different growth stages of both cultivations. The time courses furthermore show that the CO₂ uptake of the plants was strongly affected by the drought which lasted from December 2011 to January 2012. For the growth cycle of maize (216 days), the NEE was ?240 g C m^?2 and for the growth cycle of soybean (154 days) ?231 g C m^?2. In order to obtain the NEE of a complete agricultural cycle (from harvest to harvest), we also considered the NEE of autumn and winter 2011. Uncertainties of the spatially partitioned NEE are quantified and discussed.