Laboratory studies were performed to determine if the addition of iron oxides throughout biosand filter (BSF) media would increase virus removal due to adsorption.The proposed mechanism is electrostatic adsorption of negatively charged virion particles to positively charged iron oxides formed during the corrosion of zerovalent iron. Initial tests conducted using continuous flow, small-scale glass columns showed high MS2 bacteriophage removal in an iron-amended sand column (5-log 10 ) compared to a sand only column (0.5-log 10 ) over 20 pore volumes. Additionally, two experiments with a column containing iron particles revealed 4 and 5 log 10 removal of rotavirus in the presence of 20 mg/L total organic carbon. Full-scale BSFs with iron particles removed >4-log 10 MS2 for the duration of the experiment (287 days), while BSF with steel wool removed >4-log 10 MS2 for the first 160 days. Plug flow for the BSF was shown to depend on uniformity between the iron oxide material and sand media grains. The results suggest that the duration of effective virus removal by iron-amended biosand filtration depends on source water conditions and the quantity and composition of iron material added. Overall, this study provides evidence that iron-amended BSFs may advance the field of point-of-use technologies and bring relief to millions of people suffering from waterborne diseases.iii ACKNOWLEDGMENTS

The use of high-throughput sequencing technologies with the 16S rRNA gene for characterization of bacterial and archaeal communities has become routine. However, the adoption of sequencing methods for eukaryotes has been slow, despite their significance to natural and engineered systems. There are large variations among the target genes used for amplicon sequencing, and for the 18S rRNA gene, there is no consensus on which hypervariable region provides the most suitable representation of diversity. Additionally, it is unclear how much PCR/sequencing bias affects the depiction of community structure using current primers. The present study amplified the V4 and V8-V9 regions from seven microalgal mock communities as well as eukaryotic communities from freshwater, coastal, and wastewater samples to examine the effect of PCR/sequencing bias on community structure and membership. We found that degeneracies on the 3= end of the current V4-specific primers impact read length and mean relative abundance. Furthermore, the PCR/sequencing error is markedly higher for GC-rich members than for communities with balanced GC content. Importantly, the V4 region failed to reliably capture 2 of the 12 mock community members, and the V8-V9 hypervariable region more accurately represents mean relative abundance and alpha and beta diversity. Overall, the V4 and V8-V9 regions show similar community representations over freshwater, coastal, and wastewater environments, but specific samples show markedly different communities. These results indicate that multiple primer sets may be advantageous for gaining a more complete understanding of community structure and highlight the importance of including mock communities composed of species of interest. IMPORTANCEThe quantification of error associated with community representation by amplicon sequencing is a critical challenge that is often ignored. When target genes are amplified using currently available primers, differential amplification efficiencies result in inaccurate estimates of community structure. The extent to which amplification bias affects community representation and the accuracy with which different gene targets represent community structure are not known. As a result, there is no consensus on which region provides the most suitable representation of diversity for eukaryotes. This study determined the accuracy with which commonly used 18S rRNA gene primer sets represent community structure and identified particular biases related to PCR amplification and Illumina MiSeq sequencing in order to more accurately study eukaryotic microbial communities.T he use of high-throughput sequencing technologies (1, 2) has transformed the field of microbial ecology by contributing to a significant body of work that has changed our understanding of microbially diverse populations in a range of ecosystems. This is particularly true for investigations of bacterial and archaeal communities that target the 16S rRNA gene (1, 3-5). However, amplicon sequencing approaches for eukaryotes hav...

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The negative energy balance of wastewater treatment could be reversed if anaerobic technologies were implemented for organic carbon oxidation and phototrophic technologies were utilized for nutrient recovery. To characterize the potential for energy positive wastewater treatment by anaerobic and phototrophic biotechnologies we performed a comprehensive literature review and analysis, focusing on energy production (as kJ per capita per day and as kJ m(-3) of wastewater treated), energy consumption, and treatment efficacy. Anaerobic technologies included in this review were the anaerobic baffled reactor (ABR), anaerobic membrane bioreactor (AnMBR), anaerobic fluidized bed reactor (AFB), upflow anaerobic sludge blanket (UASB), anaerobic sequencing batch reactor (ASBR), microbial electrolysis cell (MEC), and microbial fuel cell (MFC). Phototrophic technologies included were the high rate algal pond (HRAP), photobioreactor (PBR), stirred tank reactor, waste stabilization pond (WSP), and algal turf scrubber (ATS). Average energy recovery efficiencies for anaerobic technologies ranged from 1.6% (MFC) to 47.5% (ABR). When including typical percent chemical oxygen demand (COD) removals by each technology, this range would equate to roughly 40-1200 kJ per capita per day or 110-3300 kJ m(-3) of treated wastewater. The average bioenergy feedstock production by phototrophic technologies ranged from 1200-4700 kJ per capita per day or 3400-13 000 kJ m(-3) (exceeding anaerobic technologies and, at times, the energetic content of the influent organic carbon), with usable energy production dependent upon downstream conversion to fuels. Energy consumption analysis showed that energy positive anaerobic wastewater treatment by emerging technologies would require significant reductions of parasitic losses from mechanical mixing and gas sparging. Technology targets and critical barriers for energy-producing technologies are identified, and the role of integrated anaerobic and phototrophic bioprocesses in energy positive wastewater management is discussed.