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Acid mine drainage (AMD), a result of oxidation of minerals containing reduced forms of sulfur (pyrites, sulfides) upon exposure to water and oxygen, is an environmental problem associated with abandoned mined lands (AML). Numerous studies have documented the impacts of AMD upon aquatic communities within acidified stream reaches; these impacts include reduced taxonomic richness and abundance, and/or a shift from pollution sensitive to pollution tolerant species. This dissertation comprises a number of integrative assessments and experiments conducted to investigate the nature of AMD ecotoxicity in the upper Powell River watershed. Emphasis was placed upon bioassessment methodologies and AMD impacts beyond the zone of pH depression. Major findings and processes developed included: 1) an Ecotoxicological Rating (ETR) system was developed that integrates chemical, toxicological, and ecological data into a single value depicting the relative environmental integrity of a given station within a watershed; 2) water column chemistry rather than sediment toxicity was the major factor causing acute toxicity to aquatic biota in close proximity to AMD discharges; 3) solid ferric hydroxide can cause acute toxicity to standard test organisms in the absence of dissolved iron; 4) Asian clams (Corbicula fluminea) can be used to detect both acutely toxic AMD inputs and nutrient loading in low order streams, and clam responses of survival and growth reflect those of indigenous communities to the two contaminant types; 5) aluminum (Al) in transition from acidic to neutral pH waters can cause acute toxicity to aquatic invertebrates, and may be the cause of impaired benthic macroinvertebrate communities in neutral pH (>7.0) waters downstream of an acidic tributary; 6) in the larger river system (North Fork Powell and Powell mainstem), urban inputs appear to have a greater influence upon aquatic communities than metal loading from AMD impacted tributaries; 7) the use of individual level assessment endpoints, such as Asian clam growth in in situ toxicity tests, eliminates variables that may confound attribution of community level impacts to contaminants; and 8) the near elimination of predatory stoneflies (Plecoptera) downstream of the Stone/Straight Creek tributary to the North Fork Powell River was associated with water column Al concentrations. This research was funded by the Virginia Department of Mines, Minerals, and Energy, Division of Mined Land Reclamation, and by the Powell River Project. / Ph. D.

Black Creek is a small watershed located in Wise County, Virginia, west of the town of Norton. At the time of this survey, the watershed encompassed approximately 929 hectares of mine and forest lands with a small recreational area. Black Creek proper is a third-order stream approximately 6.7 km in length from its headwaters to its confluence with the Powell River in Kent Junction. Black Creek and several of the tributaries within the watershed were previously identified as areas impacted by acid mine drainage. The watershed was used in a study to identify sources of acid mine drainage and the best methods for its evaluation. The acid mine drainage sources were first identified using visual inspection and field chemistry. Additional stream segments were then included in the assessment process using metal (aluminum, copper, iron, magnesium, manganese, and zinc) analyses of both overlying water column and sediments. Using an upstream reach of Black Creek as a reference, short-term toxicity testing was employed, as well as a long-term purge study. The pH at sampling locations ranged from 2.75 to 7.87 SU, and conductivity ranged from 196 μmhos/cm to 2040 μmhos/cm. All metals were elevated when compared to the reference. Water column samples collected from locations with low pH were acutely toxic to Daphnia magna and Pimephales promelas. Mortality was high in the elutriant test at locations where pH was low, conductivity was elevated, metals were high, or a combination of these. In the initial sediment tests, all sampling locations were significantly different than the reference for survival of Chironomus tentans and reproduction of D. magna. One location was significantly different than the reference for survival of D. magna. In the sediment tests completed after two months, survival of C. tentans was only different from the control in three locations but was significantly different for growth at all locations. Reproduction by D. magna was again significantly less than the reference at all locations. At eight months, only two locations were significant for survival of C. tentans and after 15 months, no significant differences occurred between any stations. The study indicates that stream segments that are severely impaired by acid mine drainage are easy to identify using visual inspection and field water chemistry. Those that are moderately impaired require more investigation and may not be responsive to short-term toxicity tests. Benthic macroinvertebrates, leaf packs, and periphyton were evaluated in the field. Benthic macroinvertebrate communities and leaf-pack breakdown were evaluated at nine locations, while periphyton was evaluated at the mouth of Black Creek, as well as five sites in the Powell River receiving system. While leaf-pack information and benthic macroinvertebrate samples yielded similar information, benthic sampling was much simpler and less time consuming. Additionally, benthic macroinvertebrate sampling, particularly over several sampling events, was more sensitive at the most severely impacted AMD stations. The stations were broken down into five different categories in order to better determine which evaluation techniques were most sensitive and cost-effective. The five categories were Non-Impaired, Slightly Impaired, Moderately Impaired, Severely Impaired, and Severely pH impaired. Once the locations were categorized, each method used to evaluate toxicity was examined to determine which methods best identified acid mine drainage impairment in the Black Creek watershed. The methods utilized include the following: basic water chemistry; metals analysis of sediments and water column; acute toxicity testing using both D. magna and P. promelas; short-term elutriant and sediment tests; chronic sediment test using C. tentans and D. magna; a purge study; benthic macroinvertebrate sampling; leaf-pack and algal-tile studies. After evaluating these methods, it was determined that using basic water chemistry and benthic macroinvertebrate sampling were the best methods for evaluating acid mine drainage impairment in this watershed. The reference station was identified as Non-Impaired. Two stations located in the lower portions of Black Creek (L11 and L1) were also Non-Impaired or only Slightly Impaired with the benthic macroinvertebrate results indicating little impairment. Stations U2, U6, U7, and BBM were also found to be Slightly Impaired. The station on the margin of the wetland, U5, was Moderately Impaired. Two previously identified areas of impairment, U9 and U10, (Cherry et al. 1995) were identified as Severely pH Impaired and Severely Impaired, respectively. / Master of Science

Thesis (M.Sc.)--University of the Witwatersrand, Faculty of Science, School of Animal, Plant and Environmental Sciences, 2017. / Acidification of freshwater systems by Acid Mine Drainage (AMD) is a persistent risk to aquatic ecosystems in South Africa, particularly in Gauteng and Mpumalanga. From several studies that have been conducted, it is clear that AMD has profound effects on aquatic life and functionality of the ecosystem. One of the ecosystem processes affected by AMD is the decomposition process. It has been established that AMD-affected streams inhibit the decompositon process. [Abbreviated Abstract. Open document to view full version] / LG2018

Thesis (MSc., Geology) -- Queen's University, 2008. / Includes bibliographical references.

Please read the abstract in the section, 00front, of this document / Dissertation (MSc (Microbiology))--University of Pretoria, 2008. / Microbiology and Plant Pathology / unrestricted

Acid mine drainage is of growing concern for both developing and developed economies. Thus there is increasing pressure to develop alternative remediation strategies. Biological sulphidogenic mechanisms have long since been studied but, very few have been implemented on a large scale. Limitations are due to the inability to acquire a suitable, low cost, environmentally friendly, renewable carbon source. The present study investigated the use of an algae biomass generated by the HRAOP of an IAPS as a carbon source for the EBRU 00AB/06 SRB consortium. The algae biomass and consortium were utilized together to remediate simulated AMD. Remediation involved decreasing the sulphate and metal concentrations in solution and decreasing the acidity of a simulated AMD. Experiments were carried out to investigate the capability of the EBRU 00AB/06 SRB consortium for sulphate reduction and sulphide generation. The consortium produced colonies when grown under anaerobic conditions in Petri dishes containing modified lactate SRB medium. The SRB consortium reduced the sulphate concentration of modified Postgates medium B and generated sulphide. Further analysis of the EBRU 00AB/06 SRB consortium revealed that the consortium was minimally impacted at pH 5 and by sulphate and iron at 3 g.L-1 and 0.5 g.L-1 respectively. The EBRU 00AB/06 SRB consortium was exposed to Actinomycin D and Ethidium Bromide to determine whether transcription and translation of proteins was required for sulphate reduction. Results indicated that sulphide generation and sulphate reduction were inducible. Analysis of the algae biomass used in this study revealed the empirical formula C1.0H1.91N0.084S0.003O0.36 indicating a carbon source rich in the nutrients required to sustain microbial development. Light microscopy revealed that algae cell walls and in particular those of Pediastrum were susceptible to acid hydrolysis. Dinitrosalicylic acid, Nile red, Bradford and Ninhydrin assays were used to determine the reducing sugar, lipid, protein and amino acid content respectively, of the mixed algae biomass. Results showed that upon exposure of the biomass to simulated AMD at pH 1 and pH 3, the concentration of reducing sugars and amino acids in solution increased. Whereas levels of lipids remained unchanged while the protein concentration decreased, indicating that, upon exposure of algae biomass to AMD, simulated or otherwise, cells ruptured, proteins were hydrolyzed and polysaccharides were broken down to sugars which are immediately available for SRB utilization. Exposure of biomass to simulated AMD revealed further that the presence of algae biomass increased the pH of simulated AMD (pH 3) to pH 7.67 after 4 d. Likewise, the pH of simulated AMD at 1 increased to 1.77 after 2 d while pH of the neutral control increased to 8.1 after 4 d. A direct comparison between lactate and algae biomass revealed 94 % sulphate removal after 23 d in the presence of algae biomass while 82 % sulphate removal was measured in the presence of lactate. Thus the EBRU 00AB/06 SRB consortium successfully utilized algae biomass for sulphate reduction and sulphide generation. In another experiment to establish if the consortium could remediate simulated AMD (pH 5) containing 0.5 g.L-1 iron and 3 g.L-1 sulphate while utilizing an algae biomass as the carbon source no residual iron was detected after 14 d and by day 23, an 89.07 % reduction in sulphate was measured. The results of this investigation are discussed in terms of utilizing a readily available and renewable biomass in the form of microalgae produced in HRAOPs as an effective carbon source in the SRB catalysed remediation of AMD.

published_or_final_version / Environmental Management / Master / Master of Science in Environmental Management

Dissimilatory sulphate reduction, brought about by the action of sulphate reducing bacteria (SRB) was used in the treatment of acid mine drainage (AMD) in a fluidised bed bioreactor. Biologically produced hydrogen sulphide and bicarbonate ions, by SRB, facilitated the precipitation of heavy metals and the generation of alkalinity in the synthetic acid mine water, respectively. The SRB that had been selected were able to utilize acetate as the sole carbon source and were capable of growing in the bioreactors at low pHs, facilitating an increase in the influent pH from 2.75-7.0 to 5.4-7.8, after a 24-hour hydraulic retention time (HRT). The precipitation efficiencies for Fe, Mn, Zn, Cu, Cr and Al after a HRT of 24 h as metal sulphides ranged between 84- 99% for influent pH values of between 4 and 7, and above 54% for influent pH values between 2.75 and 4. Microbial metabolic activity decreased with decreasing influent pH. This was inferred from the decreasing differences in chemical oxygen demand (COD) depletion rate over a 24 h HRT, as influent acidity levels approached pH 2.75. Molecular studies, using PCR-DGGE analysis on the microbial consortium in the bioreactor, revealed the presence of at least 8 different bacterial species in the consortium. Attempts at sequencing these bands yielded inconclusive results, with the bands showing sequence homology to a large number of previously uncultured and undescribed bacteria. Scanning electron microscopy confirmed the presence of bacteria of different morphology, as well as the presence of biofilms, which account for the heavy metal and low pH tolerances that the bacteria sustained.