<p>This thesis is an attempt to describe how different environmental factors influence life history traits in different populations, sexes and developmental stages in the moor frog, <i>Rana arvalis. </i>The studied populations are located along 1100 km latitudinal gradient, with pH varying between 4.0 and 8.5. I have used data from both natural populations and common garden experiments. </p><p>Reproducing moor frogs were larger and older at high latitudes, indicating a selective advantage of large size at high latitudes and/or earlier reproduction at low latitudes. When controlling for age I found that frogs were older and smaller at low pH, which may be a result of a reduced growth rate due to acid stress. The both sexes respond differently to different environments, with the lowest sexual dimorphism in body size found in the acid environments. This is possibly caused by a trade-off between growth and reproduction. Being large is considered to be advantageous, in females due to increased fecundity, and in males due to higher ability to compete for mates, while the cost of high growth is a reduced possibility to survive until the next mating season. Moor frog embryos originating from an acid population survived better under acid stress than embryos from a neutral population. Using quantitative genetic techniques I found strong maternal effects and small additive genetic variation for the traits in acid and non acid populations. The variation in acid stress tolerance owed largely to non-genetic effects. Females from acid localities lay larger eggs, which probably improves the performance of tadpoles under acid conditions. The trade-off between egg size and fecundity was stronger in acid populations indicating that females in acid populations reduced fecundity to increase offspring size. Finally, frogs from acidified environments were more asymmetric in skeletal traits further indicating the developmental stress created by acidification.</p>

Proton exchange membrane (PEM) fuel cells are an eco-friendly power source that has great potential to reduce our oil dependence for our stationary and transportation applications. In order to make PEM fuel cells an economically viable option, further effort is needed to improve proton conduction under wide operating conditions and reduce the cost of production. Design and synthesis of novel membranes that have superior characteristics require a fundamental molecular-level understanding of the relationship between the polymer chemistry, water content and proton conduction. The performance of a fuel cell is influenced by the electrochemical and molecular/proton transport processes that occur at the catalytic sites in the electrode/electrolyte interface. Therefore, understanding the molecular-level details of proton transport and structure of the multi-phase interfaces is critical. This work is subdivided into two main tasks. The first task is to model membrane/water vapor interfaces and to study their morphology and the transport properties of water and hydronium ions. Classical molecular dynamics simulation is used as the modeling tool for the characterization of the interface. The second task is to model proton transport through the aqueous domains of PEM. Such a model is inherently challenging since proton transport occurs through a combination of structural and vehicular diffusions that are associated with disparate time scales. Toward this end, we have developed and implemented a new reactive molecular dynamics algorithm to model the structural diffusion of proton that involves breaking and forming of covalent bonds. The proton transport through aqueous channels in PEM is governed by acidity and confinement. Therefore, systems in which the acidity and confinement can be independently varied, including bulk water, aqueous hydrochloric acid solutions and water confined in carbon nanotubes are also examined in addition to the application in PEM. We have developed an understanding of how acidity and confinement independently impact proton transport. The correlation between the two components of charge diffusion and their contribution to the total charge diffusion has also been explored for a basic understanding of the proton transport mechanisms. These studies will eventually help us establish the correlation between the morphology of the membrane and proton conduction.

This thesis is an attempt to describe how different environmental factors influence life history traits in different populations, sexes and developmental stages in the moor frog, Rana arvalis. The studied populations are located along 1100 km latitudinal gradient, with pH varying between 4.0 and 8.5. I have used data from both natural populations and common garden experiments. Reproducing moor frogs were larger and older at high latitudes, indicating a selective advantage of large size at high latitudes and/or earlier reproduction at low latitudes. When controlling for age I found that frogs were older and smaller at low pH, which may be a result of a reduced growth rate due to acid stress. The both sexes respond differently to different environments, with the lowest sexual dimorphism in body size found in the acid environments. This is possibly caused by a trade-off between growth and reproduction. Being large is considered to be advantageous, in females due to increased fecundity, and in males due to higher ability to compete for mates, while the cost of high growth is a reduced possibility to survive until the next mating season. Moor frog embryos originating from an acid population survived better under acid stress than embryos from a neutral population. Using quantitative genetic techniques I found strong maternal effects and small additive genetic variation for the traits in acid and non acid populations. The variation in acid stress tolerance owed largely to non-genetic effects. Females from acid localities lay larger eggs, which probably improves the performance of tadpoles under acid conditions. The trade-off between egg size and fecundity was stronger in acid populations indicating that females in acid populations reduced fecundity to increase offspring size. Finally, frogs from acidified environments were more asymmetric in skeletal traits further indicating the developmental stress created by acidification.

Synchrotron-based sulfur spectroscopy reveals a common concern for marine archaeological wood from seawater: accumulation of reduced sulfur compounds in two pathways. The distribution of sulfur species in the oak wood cell structure was mapped by scanning x-ray spectro-microscopy (SXM). Organically bound sulfur was found within lignin-rich parts, identified mainly as thiols and disulfides by sulfur K-edge x-ray absorption near edge structure (XANES) spectroscopy. Particles of iron sulfides, which may form in the presence of corroding iron, appeared in wood cavities. Cores scanned by x-ray fluorescence (XRF) show that high sulfur accumulation is restricted to the surface layers for the Swedish shipwreck Vasa, while the distribution is rather uniform throughout the hull timbers of the Mary Rose, U.K. Laboratory experiments, exposing fresh pine to simulated seabed conditions, show that the organically bound sulfur develop in reactions between lignin, exposed by cellulose-degrading erosion bacteria, and hydrogen sulfide produced in situ by scavenging sulfate reducing bacteria. With bacteria inoculated from shipwreck samples also iron sulfides formed. The iron sulfides oxidise in high humidity, and are the probable main cause of the numerous outbreaks on the Vasa’s hull of acidic sulfate salts, which were identified by x-ray powder diffraction (XRD). The iron ions catalyse several wood-degrading oxidative processes. Multi-elemental analyses were performed by scanning electron microscopy (SEM) and x-ray photoelectron spectroscopy (ESCA). The present amounts of total S remaining in the Vasa and the Mary Rose are estimated to at least 2 tonnes. After the Vasa´s spray treatment with polyethylene glycol solutions ceased in 1979, the continuing oxidation processes are estimated to have produced 2 tonnes of sulfuric acid in the wood. Laboratory experiments to gently neutralize acidic Vasa wood by ammonia gas have been conducted with promising results.

The effects of the additions of some commonly-available organic residues to an acid, P-deficient soil (typical of those used by small-scale farmers in KwaZulu-Natal) on soil pH, exchangeable and soil solution AI, P availability and maize yield response was investigated in a number of laboratory and glasshouse experiments. The organic amendments used were ground-up grassveld residues, household compost, filter cake (a waste product from a sugar mill) and layer poultry manure. The soil used was a Hutton form (Farmingham series) (Rhodic Ferrasol, FAO). In an initial laboratory study, addition of all of the organic residues, at rates equivalent to 10 and 20 Mg ha¯¹, raised soil pH significantly and as a result there was a marked reduction in exchangeable AI concentrations. The increase in pH and decrease in exchangeable AI was more pronounced at the higher rate of addition and followed the order: poultry manure> filter cake> household compost> grass residues. The major mechanism responsible for the increase in pH was thought to differ depending upon the type of organic residue being considered. Whilst the relatively high content ofCaC0₃ was probably the main mechanism in the case of poultry manure and filter cake, the proton consuming ability of humic material probably predominated for household compost and decarboxylation of organic acids during decomposition was probably the main mechanism in the case of grass residues. Additions of organic amendments also decreased concentrations of total AI (AIT) in soil solution but the concentration of monomeric AI (AIMono) as estimated by pyrocatechol violet 60 sec. method, was unchanged or even increased. This latter effect was attributed to the high cation content of residues (particularly that of poultry manure) which increased soil salinity and exchangeable AI³⁺ was consequently displaced into soil solution.Additions of amendments also increased the Olsen-extractable P levels in the order: poultry manure> filter cake> household compost> grass residues and their addition also decreased theP adsorption capacity ofsoils. Concentrations of exchangeable Ca, Mg and K, and Na in the case of poultry manure, were increased by additions of organic amendments. In a glasshouse experiment, the four organic residues were applied to soils at a rate equivalent to 20 Mg ha¯¹ with or without the addition of either lime (equivalent to 0, 5 or 10 Mg ha¯¹) or P (equivalent to 0, 10 or 50 kg ha¯¹). Lime applications to the control (unamended) treatment resulted in a marked reduction in exchangeable AI, AIT AIMono and in the proportion of AIT present as AIMono in soil solution. The addition of organic amendments increased soil pH and reduced AIT and AIMono to low concentrations regardless of whether lime was applied or not. There was no yield response in maize to applied lime in any of the amended treatments. There was a yield increase in response to applied P in the control, household compost and grass residue treatments but none for the filter cake and poultry manure treatments. In agreement with this, Olsen-extractable P values in soils followed the order: poultry manure> filter cake> household compost> grass residues > control. It was concluded that the addition of organic amendments to acid soils is a practicable way of liming them and reducing the potential for Al toxicity and that it can also reduce fertilizer P requirements. This research now needs to be extended into the field situation. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2000.

This study describes two projects involving recent research on atmospheric ammonia. The first project investigates differences in modelling techniques of aerosol acidity using data from two recent field campaigns. Our results show that allowing or disallowing gas-particle partitioning in the Extended Aerosol Inorganic Model (E-AIM) changed the average modelled aerosol activity of H+ from one campaign by seven orders of magnitude and that disallowing gas-particle partitioning may not accurately represent the chemical state of the aerosols. The second project investigates the interference of reduced nitrogen in commercial chemiluminescent nitrogen oxide monitors with molybdenum oxide catalytic converters. This phenomenon is strongly dependent on the temperature of the catalytic converter. Our results show these instruments can have high conversion efficiencies of gaseous NH3 and NH4+ salts to NO at typical reported converter temperatures, but conversion efficiency varies between instruments and may be the result of uncertainty in reported converter temperature.

Laboratory studies and field trials were carried out to investigate the effect of addition of some organic residues to acid soils on soil pH, exchangeable and soluble AI, nutrient status, microbiological and biochemical indices and maize response. The organic wastes used in the first laboratory study included plant materials (maize. sorghum, kikuyu grass, soybean, red clover residues and acacia prunings), animal manures (kraal, pasture-fed and feedlot cattle manure, layer and broiler poultry manure and pig manure), household compost, sewage sludge, and filter cake. The poultry manure, pig manure and leguminous plant residues had the highest content of basic cations while sewage sludge had the highest N content. Poultry manure had very high values for proton consumption capacity, CaC03 content and ash alkalinity. Proton consumption capacity, ash alkalinity, total basic cation content and CaC03 content were closely correlated with one· another. Soil pH was increased and exchangeable AI and total (AIT) and monomeric (AIMono) AI in solution were decreased by addition of all the organic wastes: the effect was greater at the higher rate of application. Strong correlations were recorded between the rise in soil pH and proton consumption capacity, ash alkalinity, CaC03content and basic cation content of the residues. The major mechanisms responsible for the elevations in pH were suggested to be the substantial CaC03content of poultry and pig manures, and filter cake, the proton consumption capacity of humic material present in household compost and manures and decarboxylation of organic acid anions during the decomposition of plant residues and manures. It was proposed that ash alkalinity is a suitable laboratory test for predicting the potential Iiming effect of organic residues since it is strongly correlated with the rise in pH that occurs, it is relatively simple to measure and the values reflect the initial content of organic acid anions, humic materials and CaC03in the residues. A preliminary field experiment was set up to investigate the effectiveness of kraal manure as a Iiming material in an acid soil (pHwater =4.1) at a site close to a Zulu village. The experiment consisted of two rates of lime (L1 = 2.5 and L2 = 5.0 t ha1) and two rates of kraal manure (K1 = 10 and K2 = 20 t ha-1 ) which were banded and incorporated in a 30 cm wide strip down the plant rows. Treatments were arranged in a randomized block design with three replicates. A commercial maize cultivar PAN 6710 and a traditional variety EMBO, used by the farmers in the locality, were grown. Soils in the plant row were sampled at tasselling and at harvest. The addition of kraal manure significantly raised soil pH and reduced concentrations of exchangeable AI and those of both total and monomeric AI in soil solution. Lime raised pH and the pH continued to increase between tasselling and harvest. Maize yields for control, kraal manure (K1 and K2) and lime (L1 and L2) for PAN 6710 were 2.5, 3.7, 5.1, 5.3 and 6.3 t ha-1 ; respectively and for EMBO they were 3.0, 5.4, 5.8, 5.9and 8.2 t ha-1 , respectively. These results demonstrate the high yield potential of the traditional maize variety under small scale farming conditions, and show that large yield increases can be obtained by applying kraal manure. The long-term effects (24 weeks) of incubation of organic wastes (soybean residues, poultry, pig and kraal manures and sewage sludge) with an acid soil were investigated in a laboratory study. After incubation for six weeks incubation, soil pH was raised and exchangeable AI and Air and AIMono in soil solution were decreased . by addition of the wastes. Soil pH generally declined and exchangeable and soluble AI increased over the remainder of the incubation period. The decline in pH was attributed mainly to nitrification of NH4+ originating from mineralization of wastes-derived organic N. Addition of organic materials generally resulted in a decrease in the proportion of solution Air present as AIMono. That is, the effects of addition of organic materials was two-fold; an increase in pH in the short term and complexation of AI by organic matter. Since these effects occur simultaneously, it would be desirable to separate them. For this reason, short-term equilibration experiments (3 days) were conducted to study the solubility of AI in aqueous solution or in an Oxisol when in equilibrium , with 3 manures (kraal, pig and poultry) at pH values of 4.0, 4.5, 5.0, 5.5, 6.0 and 6.5. Addition of manures tended to reduce the concentrations of total AI in solution (AIT) in the lower pH range (Le. pH 4.0 and 4.5) but increased AIT concentrations compared to the control, at higher pH values (Le. at pH 5.5 and above). This was explained in terms of the complexing ability of both the solid and solution phases. At lower pH, where AI is highly soluble, complexation by added solid phase manure-organic matter results in a reduction of AI solubility. However, at high pH, where AI solubility is limited, the most important mechanism is complexation of AI by soluble organic matter and this increased AI solubility. Additions of manure reduced the proportion of Air present in monomeric form (AIMono). This effect was more pronounced in aqueous solution but was also clearly evident above pH 5.0 in the Oxisol. This reflects the fact that a large concentration of soluble C in solution can maintain relatively high concentrations of complexed AI in solution but at the same time maintain low concentrations of AIMono. It was concluded that formation of AI-organic matter complexes caused by additions of organic manures can alter the solubility of AI and reduce the amount of phytotoxic AIMono present in soil solution. A second field trial was conducted to compare the effects of additions of kraal manure, grass residues, lime and fertilizer (N-P-K) under field conditions, on soil pH, AI solubility and maize response and, at the same time follow concomitant changes in the size and activity of the soil microbial biomass and enzyme activity. The greatest effects of kraal manure in increasing soil pH and decreasing AI toxicity were recorded six weeks after planting whereas those of lime and grass residues were recorded at harvest. Kraal manure and fertilizer increased significantly AMBIC extractable P and exchangeable K and Zn. In addition, Kraal manure, and to a lesser extent lime significantly increased exchangeable Ca and Mg.. Soils in the plant row in the grass residue treatments had the highest microbial biomass C and microbial quotient,followed by kraal manure, lime and controls. Basal respiration rates and arginine ammonification, protease, aryl sulphatase, and acid phosphatase activity rates were significantly increased by addition of all treatments and these increases tended to be accentuated by fertilizer. Low values for metabolic quotient in the grass residue treatments were associated with high values for microbial biomass C in these treatments. The addition of all treatments tended to increase maize yields and, in general, these yields were greater for the high rate of application of each amendment. Yields for unfertilized kraal manure were markedly greater than those for the unfertilized grass residue and lime treatments. This was attributed to the ability of kraal manure to both increase pH and add nutrients to the soil. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

This study describes two projects involving recent research on atmospheric ammonia. The first project investigates differences in modelling techniques of aerosol acidity using data from two recent field campaigns. Our results show that allowing or disallowing gas-particle partitioning in the Extended Aerosol Inorganic Model (E-AIM) changed the average modelled aerosol activity of H+ from one campaign by seven orders of magnitude and that disallowing gas-particle partitioning may not accurately represent the chemical state of the aerosols. The second project investigates the interference of reduced nitrogen in commercial chemiluminescent nitrogen oxide monitors with molybdenum oxide catalytic converters. This phenomenon is strongly dependent on the temperature of the catalytic converter. Our results show these instruments can have high conversion efficiencies of gaseous NH3 and NH4+ salts to NO at typical reported converter temperatures, but conversion efficiency varies between instruments and may be the result of uncertainty in reported converter temperature.

Acid mine drainage (AMD) is a significant environmental issue worldwide, which often causes severe contamination and marked species losses in receiving streams. However, little is known about how this stress alters food webs and ecosystem function. I conducted a literature review, which revealed that AMD-impacted streams generally had depauperate benthic communities dominated by a few tolerant species and impaired ecosystem processes. Next, using survey and experimental-based approaches, I investigated food web structure and energy flow in these highly stressed streams, which typically have low pH (< 3), high concentrations of dissolved metals (Al, Fe), and substrata coated with metal hydroxide precipitates, on the South Island, New Zealand. Inputs of AMD caused substantial loss of consumers and reduced the overall number of links between species generating small and simplified food webs, with few invertebrates and no fish. Comparative analysis of food webs from a survey of 20 streams with either anthropogenic or natural sources of acidity and metals, indicated that anthropogenic sources had a stronger negative effect on food web properties (size, food chain length, number of links); an effect driven primarily by differences in consumer diversity and diet. However, the presence of fewer trophic levels and reduced trophic diversity (detected using isotopic metrics), were common structural attributes in AMD-impacted webs along a pH gradient, regardless of impact level. Furthermore, complementary dietary analyses of consumer gut contents and stable isotope signatures (δ13C and 15N) confirmed that primary consumers fed generally on basal resources and that there were few predatory interactions, which reflected low densities of small-bodied chironomids. This suggests that food quantity was unlikely to limit primary consumers but that reduced prey availability may be an additional stressor for predators. In these radically re-structured food webs, trophic bottlenecks were generated at the primary consumer level and energy flow to higher consumers was disrupted. However, streams still retained some limited function, including slow leaf litter breakdown, which provided detrital resources and supported the small food webs. Overall, my findings have furthered our understanding of these highly stressed stream ecosystems by providing new insights into interactions among species and trophic levels that structure food webs and enable function.

Acid mine drainage (AMD) is a major environmental issue worldwide. On the West Coast of the South Island, New Zealand, numerous catchments receive AMD, with significant negative impacts on in-stream flora and fauna. Diatoms are commonly regarded as powerful biological indicators and may be found in high abundance in AMD-contaminated streams; however, relatively little work has been done on diatoms in mining environments in New Zealand. Initially, I conducted a survey of epiphytic diatom communities in 39 streams ranging from non-impacted reference streams to those severely impacted by AMD. Streams were assigned to one of four classes along an AMD gradient: circum-neutral reference, naturally acidic reference, moderately impacted, and severely impacted. There was a wide range in diatom taxonomic richness in reference and moderately impacted streams (8 – 33 taxa). Taxonomic richness was greatly reduced in severely impacted streams (1 – 5 taxa) at a threshold of pH 3.4 and was dominated by Pinnularia cf. acidophila (69 – 100% relative abundance). Community composition differed between circum-neutral reference, moderately, and severely impacted streams; however, naturally acidic and moderately impacted streams had similar diatom communities primarily composed of acid-tolerant Eunotia and Frustulia species. This indicated that diatoms are strongly structured by pH and able to tolerate moderate conductivity and metal concentrations without a corresponding shift in community composition. Survey data were then used to develop two diatom-based indices for streams impacted by AMD: a single Biotic Index and a Multimetric Index. While neither index was able to distinguish naturally acidic from moderately impacted streams, both indices successfully categorised streams as circum-neutral reference, moderately or severely impacted by AMD. These indices may be useful in assessing AMD impact on circum-neutral streams or in identifying when a stream has crossed a threshold from moderately to severely impacted by AMD. Diatoms would be especially useful as bioindicators of AMD if they respond rapidly to a change in mine discharge. To test this, mature algal biofilms were reciprocally transferred between circum-neutral reference streams and streams of varying degrees of AMD over a period of 13 days. Diatom mortality increased rapidly in the reciprocal transfer between reference and severely impacted streams. Reference communities resembled the ambient diatom community of severely impacted streams 13 days post-transfer. However, in the reverse transfer, a change in community composition was slow to occur. Diatoms respond faster to an increase in pollution than to pollution amelioration. Overall, results indicated that diatom communities may be a useful tool for monitoring the presence and magnitude of AMD in New Zealand streams.