The ultimate objective of this work was to develop a real-time method for detecting and monitoring β-D-galactosidase as a suitable indicator of the potential presence of total coliform bacteria in water environments. Preliminary comparison of the chromogenic substrate, chlorophenol red β-D-galactopyranoside and the fluorogenic substrate, MuGAL, revealed unreliable results with the fluorogenic technique due to interference from compounds commonly found in environmental water samples. Thus, the chromogenic assay was further explored. Hydrolysis of the chromogenic substrate chlorophenol red β-D-galactopyranoside by β-D-galactosidase to yield chlorophenol red was the basis of this assay. Fundamental studies with chlorophenol red β-Dgalactopyranoside showed that β-D-galactosidase occurs extracellularly and in low concentrations in the polluted water environment. A direct correlation between enzyme activity and an increase in environmental water sample volume, as well as enzyme activity with total coliform colony forming unit counts were observed. Spectrophotometric detection was achieved within a maximum period of 24 h with a limit of detection level of 1 colony forming unit 100 ml[superscript -1]. This enzyme also exhibited physical and kinetic properties different from those of the pure commercially available β-D-galactosidase. Cell permeabilisation was not required for releasing enzymes into the extracellular environment. PEG 20 000 offered the best option for concentrating β-D-galactosidase. The source of β-D-galactosidase in the polluted environmental water samples was confirmed as Escherichia coli through SDS-PAGE, tryptic mapping and MALDI-TOF, thus justifying the further use of this method for detecting and/or monitoring total coliforms. Several compounds and metal ions commonly found in environmental water samples (as well as those used in water treatment processes) did have an effect on β-D-galactosidase. All the divalent cations except Mg [superscript 2+], at the concentrations studied, inhibited the relative activity of β-D-galactosidase in both commercial β-D-galactosidase and environmental samples. Immobilisation of chlorophenol red β-D-galactopyranoside onto a solid support material for the development of a strip bioprobe was unsuccessful, even though the nylon support material yielded some positive results. A monthly (seasonal) variation in β-Dgalactosidase activity from the environmental water samples was observed, with the highest activity coinciding with the highest monthly temperatures. Electro-oxidative detection and/or monitoring of chlorophenol red was possible. Chlorophenol red detection was linear over a wide range of concentrations (0.001-0.01 μg ml[superscript -1]). Interference by chlorophenol red β-D-galactopyranoside in the reduction window affected analysis. A range of phthalocyanine metal complexes were studied in an attempt to reduce fouling and/or increase the sensitivity of the biosensor. The selected phthalocyanine metal complexes were generally sensitive to changes in pH with a reduction in sensitivity from acidic pH to alkaline pH. The tetrasulphonated phthalocyanine metal complex of copper was, however, more stable with a minimum change of sensitivity. The phthalocyanine metal complexes were generally stable to changes in temperature. While only two consecutive scans were possible with the unmodified glassy carbon electrode, 77 consecutive scans were performed successfully with the CuPc-modified glassy carbon electrode. Among the phthalocyanine metal complexes studied, the CuPc-modified glassy carbon electrode therefore provided excellent results for the development of a biosensor. The CuPc modified-glassy carbon electrode detected 1 colony forming unit 100 ml[superscript -1] in 15 minutes, while the plain unmodified glassy carbon electrode required 6 hours to detect the equivalent number of colony forming units. CoPc, ZnPc and CuTSPc required 2, 2.25 and 1.75 h, respectively, to detect the same numbers of colony forming units. The CuPcmodified glassy carbon electrode detected 40 colony forming units 100 ml[superscript -1] instantly. In general, a direct correlation between colony forming units and current generated in the sensor was observed (R2=0.92). A higher correlation coefficient of 0.99 for 0-30 coliform colony forming units 100 ml[superscript -1] was determined. Current was detected in some water samples which did not show any colony forming units on the media, probably due to the phenomenon of viable but non-culturable bacteria, which is the major disadvantage encountered in the use of media for detecting indicator microorganisms. This novel biosensor therefore presents a very robust and sensitive technique for the detection and/or monitoring of coliform bacterial activity in water.

Hatchling disorientation after emergence is a major factor impacting sea turtle populations. This study utilized data from over 1,200 Florida Fish and Wildlife Conservation Commission (FWC) Marine Turtle Disorientation Report forms from years 2006 to 2011 to assess changes in the severity and locations of disorientation events and the impact of municipal beach lighting ordinances. While the FWC forms were completed for all sea turtle species observed, this study focused only on loggerhead sea turtles (Caretta caretta). A Disorientation Severity Index (DSI) was derived from the number of hatchlings and the direction of their tracks leaving the nests to evaluate the changes in disorientation over six years in Broward County. The FWC forms provide a much larger database for the analysis of hatchling disorientation patterns and trends than can be derived from the more precise, but labor intensive, Hatchling Orientation Index (HOI) survey method of Witherington et al. (1996). This research differs from prior work by focusing on the information provided in the FWC Marine Turtle Disorientation Report forms; using each individual disorientation to assess changes in the severity of hatchling disorientations over time. Prior work has not used the FWC forms for analysis. Significant differences were found for average DSI between years and locations. Overall, DSI decreased significantly from 2006 to 2011 in Hillsboro Beach, Pompano Beach, Lauderdale-by-the-Sea, Fort Lauderdale, and Hollywood. This might be due to increased compliance with lighting ordinances. In addition, disorientation hotspots were identified and the DSI in these hotspots decreased significantly in central and south Fort Lauderdale, Pompano Beach, and Lauderdale-by-the-Sea. Hotspots were visually identified as R-Zone ranges with higher numbers of disorientations than in other ranges (Fig. 6). Artificial beach illumination is very prevalent in Broward County. However, there was no significant relationship between the number of types of lights that were recorded on the FWC forms and DSI. Disorientations seemed to be clustered within hotspots with known lighting issues. Management strategies should use these analyses to reassess loggerhead recovery plans to reduce disorientation hotspots and increase hatchling recruitment.

M.Sc.

The use of biomarkers in assessing the health of aquatic ecosystems is widely being used. Biomarkers have been proposed as sensitive early warning tools for biological measurement in environmental quality assessment. In this study, DNA damage, HSP 70 expression, Lactate dehydrogenase and alkaline phosphatase were evaluated for their usefulness and applicability in the active biomonitoring of the Rietvlei system using Melanoides tuberculata and Oreochromis mossambicuss as test organisms. Levels of DNA strand breakage were measured in the exposed test organisms and the references that were kept in the laboratory under unpolluted environment. Different levels of DNA damage were recorded in M tuberculata that were exposed during high-flow and those from the low-flow exposure period. There was no difference in DNA between the control and high-flow exposure M. tuberculata but the low-flow exposure duration had significantly higher DNA damage for all the three sites when compared to the high-flow and the controls. There was no difference in the amount of DNA damage in both exposed and control O. mossambicus. Fish samples were only available for the high-flow four weeks exposure because of the test organisms dying during the low-flow exposure. Heat shock protein 70 expression was determined in O. mossambious only and significant differences in expression between sites were demonstrated with highest expression at site 1 decreasing down stream at sites 3 and 5. The activity of LDH in M. tuberculata was not different in both control and exposed fish at all the sites. There was no difference in the activity of LDH between the high-flow and low-flow exposure groups in M. tuberculata. In O. mossambicus, the activity of LDH was significantly inhibited at site 1 with the effects of the contaminants decreasing downstream at sites 3 and 5 where LDH activity did not differ from the controls. There was no alkaline phosphatase activity in both exposed and control M. tuberculata but in O. mossambicus alkaline phosphatase was significantly inhibited at the three sites although inhibition did not differ from site to site. This suggests that LDH and alkaline phosphate are not sensitive biomarkers in M. tuberculata in the Rietvlei system but are sensitive in O. mossambicus. DNA damage was shown to be a sensitive biomarker using M. tuberculata but it was difficult to assess this in O. mossambicus since fish were only available during the 4 weeks high-flow exposure, which did not show any differences from the control. The lack of DNA damage in O. mossambicus maybe a result of low concentration of contaminants during high-flow or because this biomarker is not sensitive in fish. / Prof. J.H.J. Van Vuren

No description available.

Environmental impact assessment is a widely accepted planning tool used in environmental management. Internationally it has been adopted as a formal permitting requirement for development projects in many jurisdictions. Historically the focus has been on the pre-decision making stages of environmental impact assessment. It has, however, been widely acknowledged that post-decision environmental impact assessment follow-up is an important component in confirming initial predictions, enabling responsible adaptive management of environmental impacts and ensuring compliance with permit conditions. It is this last function which is the focus of this study. Specifically, the role of permit conditions in enabling compliance and facilitating compliance monitoring is addressed. Permit conditions of twenty-one environmental authorisations were examined and tested for conformance with legislated requirements, and practicality of monitoring for compliance (monitorability). It was found that there are many contributors to achieving monitorable permit conditions. Amongst the most significant of these are conformity in interpretation of the regulations specifying permit content by officials, gaps in guidance on the part of the regulations themselves, and a tendency to focus on construction related impacts. The lack of clarity regarding the roles and functions of environmental control officer and environmental auditor further contribute to poor monitorability of permit conditions. Specific areas of shortcoming and best practice in the permit conditions analysed were identified and discussed. Finally, recommendations are made for the improvement of permit condition monitorability.

The focus of the National Water Act (Act 36 of 1998) is on the sustainable utilization of our water resources. This is to be achieved through the implementation of an integrated resource protection approach, which is aimed at ensuring that a balance is maintained between the protection and utilization of our countries water resources. This approach sets Resource Quality Objectives (RQOs) that define acceptable levels of water resource protection. The acceptable risk of damage to the ecological integrity of a water resource will play an important role in the setting of these objectives, e.g. for a water resource of lower importance a higher risk would be acceptable with the subsequent setting of RQOs at less stringent levels. A desktop Resource Directed Measures (RDM) determination has already been performed for the Luvuvhu River in the Northern Province. There was decided to evaluate a facet of this desktop study, namely the Present Ecological Status (PES), by utilising the South African Scoring System version 4 (SASS4) and the Integrated Habitat Assessment Method (IHAS) biomonitoring techniques. It was then possible to compare the desktop determined PES to the PES determined from the information provided by the biomonitoring techniques. Ultimately, SASS4 verified the reliability of the RDM methodology Further, to facilitate the introduction of Ecological Risk Assessment (ERA) into South Africa there was looked at how and where SASS4 and IHAS would fit into the various phases of the ERA process. SASS4 serves as an indication of the extent of an impact, and in conjunction with an ERA, would provide the means with which to determine causality. A retrospective ERA based on data obtained from SASS4 and a concomitant habitat assessment method will thus provide a valuable tool for the protection of our water resources. / Prof. G.J. Steyn

A recente explosão de visitação em áreas naturais está causando o desgaste justamente dos valores que tornaram tais áreas atrativas. As mudanças na estrutura da comunidade são problemas graves, principalmente onde há falta de conhecimento prévio, como os costões rochosos. O levantamento fisionômico visa possibilitar o aumento da velocidade e abrangência na aquisição de dados, importante em estudos de avaliação de impacto. O objetivo deste trabalho foi realizar uma avaliação das alterações na estrutura da comunidade bentônica relacionadas à visitação pública. O estudo foi realizado na Piscina Natural do Parque Estadual da Ilha Anchieta, um importante destino turístico do litoral norte paulista que apresenta grande visitação durante o mês de janeiro. Amostragens foram realizadas na Piscina Natural e em duas áreas de referência, em dezembro de 2008 e no final de janeiro de 2009, seguindo o modelo BACI (befor-after-control-impact). Os dados foram analisados através de Índices de Similaridade e dendogramas. Foram encontrados 24 povoamentos. No médio-litoral o maior recobrimento foi de Banco de Chtamallus (>35%) e no infra-litoral foi de Tapete de Jania e Amphiroa (>45%). Para os dados qualitativos, a similaridade entre as áreas foi alta (>0,79), assim como entre as duas amostragens em cada área (>0,75). Para os dados quantitativos a similaridade também foi alta (>0,82). Os únicos povoamentos que apresentaram diferença significativa entre as amostragens foram Banco de Ectocarpaceae, Banco de Brachidontes e Banco de Sargassum na Área de Referência 1 e Crosta de Corallinaceae na Área de Referência 2. Os resultados indicam que a comunidade bentônica de substrato consolidado da Piscina Natural não apresentou alterações significativamente maiores que as áreas de referência nesse intervalo de tempo. Entretanto, não é possível concluir que não está ocorrendo impacto, já que o método pode não ter detectado alterações. Este foi o primeiro trabalho que se propôs a fazer uma avaliação de impacto utilizando esse método. Portanto, são necessários mais estudos para adequar o método a este objetivo. / The recent boom in visiting natural areas is destroying the very features that make those areas so attractive. Changes in the structure of natural community are serious problems, especially at poor known areas, such as rocky shores. A physiognomic assessment aims to produce an increase in data acquisition amplitude and speediness, relevant features in impact evaluation studies. Our goal was to evaluate structure changes in a benthic community related to public visitation. The study was realized at Piscina Natural in the State Park Anchieta Island, a important touristic point at northern coast of São Paulo state, which receive many tourists during January. Samplings were done at Piscina Natural and at tow reference areas, in 2008 December and at the end of 2009 January, following BACI model (before-after-control-impact). Data were analyzed using similarity index and dendrogram. Twenty four settlements were found. At midlittoral Chtamallus Bed had the highest percent cover (>35%), and at infrallittoral Jania and Amphiroa Turf had the highest percent cover (>45%). With qualitative data, similarity among areas was high (>0,79), as well as among sampling periods in the same area (>0,75). With quantitative data similarity was also high (>0,82). Only a few settlements showed significant differences between sampling periods, Ectocarpaceae Bed, Brachiontes Bed and Sargassum Bed at Reference Area 1 and Corallinaceae Crusts at Reference Are 2. This results indicate that benthic community on consolidated substrate at Piscina Natural did not undergo alteration significantly higher than those from reference areas in this time interval. However, we cannot conclude that there is no impact, the methodology could had not be able to detect the impact. This was the first study that aimed to evaluate environmental impact, using this methodology. Therefore more studies are needed to fit this methodology to this objective.

The purpose of this study is to characterize the water chemistry of Lake Gregory in Crestline, California. The collection of water samples occurred a short time after the end of the summer recreational season in the late fall. Several chemical parameters were measured in an effort to determine the overall water quality during this time.

We developed curved spiral antennas for use in underwater (freshwater) communications. Specifically, these antennas will be integrated in so-called mussel backpacks. Backpacks are compact electronics that incorporate sensors and a small radio that operate around 300 MHz. Researchers attach these backpacks in their freshwater mussel related research. The antennas must be small, lightweight, and form-fit the mussel. Additionally, since the mussel orientation is unknown, the antennas must have broad radiation patterns. Further, the electromagnetic environment changes significantly as the mussels burrow into the river bottom. Broadband antennas, such a spiral antennas, will perform better in this instance. While spiral antennas are well established, there has been little work on their performance in freshwater. Additionally, there has been some work on curved spiral antennas, but this work focused on curving in one dimension, namely curving around a cylinder. In this thesis we develop spiral antennas that curve in two dimensions in order to conform the contour of a mussel's shell. Our research has three components, namely (a) an investigation of the relevant theoretical underpinning of spiral antennas, (b) extensive computer simulations using state-of-the art computational electromagnetics (CEM) simulation software, and (c) experimental validation. The experimental validation was performed in a large tank in a laboratory setting. We also validated some designs in a pool (∼300,000 liters of water and ∼410 squared-meter dive pool) with the aid of a certified diver. To use CEM software and perform successful antenna-related experiments require careful attention to many details. The mathematical description of radiation from an antenna, antenna input impedance and so on, is inherently complex. Engineers often make simplifying assumptions such as assuming no reflections, or an isotropic propagation environment, or operation in the antenna far field, and so on. This makes experiments on antennas challenging since it often quite difficult to replicate the simplifying assumptions in an experimental setting. Still, with careful consideration of the important factors and careful experimental design it is possible to perform successful experiments. For example, antenna measurements are often performed in anechoic chambers. For our research we used a large swimming pool to mimic an underwater anechoic chamber. Our CEM simulations and experimental results are in most cases congruent. We are confident that we can design formfitting, compact (spiral) antennas that one could deploy on mussels. This will greatly enhance the mussel backpacks that are used by researchers at the University of Iowa.