2015 Symposium Abstracts - Environmental Engineering

ENE-01  Exposure To Higher Doses Of 2,3,7,8-Tetrachlorodibenzo-p-dioxin Causes Changes In Specific Bacteria Communities In Mice Gut Microbiota

Authors: Prianca Bhaduri; Robert D. Stedtfeld; Tiffany Stedtfeld; Kelly A. Fader; Norbert E. Kaminski; Timothy R. Zacharewski; James M. Tiedje; Syed A. Hashsham

Abstract: Specific bacteria in the gut including segmented filamentous bacteria (SFBs), Clostridia cluster IV, Clostridia cluster XIVa and Bacteroides fragilis are important commensals that are known to respond to changes in the intestinal micro-environment and help modulate immune responses. The environmental contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been widely studied as a potent toxin that suppresses immune responses. TCDD dose-population abundance studies were conducted to observe changes in the abundance of SFBs within the intestine of mice that have been exposed to TCDD. We studied SFB abundances before and after TCDD dosage (30 µg/kg) in mice and found an increasing trend in SFB populations after dosage. Also, fecal samples were collected weekly from mice treated with 0, 0.01, 0.03, 0.1, 0.3, 1, 3, 10 or 30 µg/kg TCDD every four days over a period of 90 days. The animals exhibited a dose-dependent increase in SFB abundance, with mice exposed to higher doses (3, 10 and 30 µg/kg TCDD) showing a significant increase in SFB numbers in their intestines. Similarly, a dose-dependent decrease in Clostridia cluster XIVa (Roseburia spp. and Eubacterium rectale), Clostridia cluster XIVa and Bacteroides fragilis were observed in both the short term and long term mice studies.

This work was supported in part by Superfund Research Program (P42ES04911) from the National Institute for Environmental Health Sciences


ENE-02  Groundwater Sustainability In Michigan Lowlands - A Multiscale Analysis

Authors: Zachary Curtis; Hua-sheng Liao; Prasanna Sampath; Shu-Guang Li

Abstract: This research investigates a coastal hazard that may significantly impact the sustainability of groundwater resources in Michigan. Recent analysis of a statewide water quality database (106+ samples) revealed that groundwater salinity in low-lying areas of the Lower Peninsula is significantly high. Moreover, this is NOT a result of surface contamination (e.g., road salt infiltration). We hypothesize that the contamination originates from below, that is to say, upwelling of brine “plumes” from deep bedrock formations is the pollution mechanism and it is driven by increased water consumption, effects of climate change, and/or land use changes. Key questions to address include: (1) how fast is the saline water moving upward (2) what is the relative importance of human pumping?; and (3) what are the management implications? Our approach systematically integrates field measurements, data-driven analysis, and process-based modeling to evaluate groundwater sustainability at multiple scales of interest. At the regional scale, we analyze large volumes of pre-processed groundwater and geo-spatial data. At the sub-regional and local scales, we collect high-quality field data and develop hierarchical groundwater models, using Ottawa County, Michigan as the study area. We recently analyzed ~450 geo-referenced groundwater samples for [Cl-] to enable water quality mapping and comparative analysis with historical Cl- levels. A county-wide groundwater model was developed to test and extend our conceptual understanding of the aquifer system. Preliminary results correlate areas with extensive subsurface clay and aggressive groundwater withdrawals to increasingly saline groundwater. Future work includes an expansion on data analysis, continued field work, and model development/application.

This work was supported in part by Ottawa County Board of Commissioners (Ottawa Michigan); Michigan Department of Agriculture and Rural Development


ENE-03  Development Of Loop Mediated Isothermal Amplification (LAMP) For Rapid Detection Of Dehalococcoides spp. In Groundwater Samples

Authors: Yogendra Kanitkar; Robert Stedtfeld; Paul Hatzinger; Rob Steffan; Syed Hashsham; Alison Cupples

Abstract: Typically, real time quantitative polymerase chain reaction (qPCR) based on the TaqMan probe or DNA binding dyes is used to quantify and monitor the in situ activity of Dehalococcoides spp. To date, a wide range of qPCR protocols to quantify 16S rRNA genes as well as reductive dehalogenase genes such as vcrA, tceA and bvcA are available. Although qPCR methods have been successful for monitoring reductive dechlorination, alternate molecular methods that are faster and cheaper, may make quantification of reductive dehalogenase genes in groundwater samples easier. In this study, we developed LAMP for rapid and specific quantification of the vinyl chloride reductive genes (vcrA and bvcA), in two commercial reductive dechlorinating SDC-9 and KB-1 as well as groundwater samples. LAMP primers specific to the vinyl chloride reductive genes (vcrA and bvcA) in Dehalococcoides spp. were designed using Primer Explorer V4. DNA templates were prepared using active KB-1 microcosms or groundwater samples from four distinct monitoring wells located on a recently SDC-9 bio augmented groundwater aquifer in Topeka, KS. Threshold times and sensitivities were determined by creating a 10 fold dilution series for both template types. Detection limits were evaluated using plasmid standards for both genes. All experiments were performed on a thermal cycler. Amplification was observed using LAMP primer sets for both KB-1 as well as groundwater templates. LAMP can detect the vinyl chloride reductive genes (vcrA and bvcA) in Dehalococcoides spp. below 107 gene copies/L, the accepted threshold for natural attenuation. The presence of PCR inhibitors in groundwater templates may explain the slightly higher detection limit compared to SDC-9 and KB-1 templates. Future research will focus on developing the LAMP assay for the detection of 16S rRNA gene as well as other reductive dehalogenase genes (pceA and tceA), identifying PCR inhibitors in groundwater and mitigating their effects on amplification.

This work was supported in part by Strategic Environmental Research and Development Program


ENE-04  Field- Based Detection Of Environmental DNA (eDNA) From Aquatic Invasive Species Using Novel Technology And Citizen Science

Authors: Maggie R. Kronlein; Robert D. Stedtfeld; Cathrine Kronlein; Erin Dreelin; Jo Latimore, R. Jan Stevenson; Syed A. Hashsham

Abstract: Environmental DNA (eDNA)-based species detection is a promising tool for monitoring and initiating early responses to aquatic invasive species (AIS) introductions. Using these genetic-based approaches, populations can be detected in much lower abundances and earlier than tradition field survey-based techniques, greatly increasing the likelihood of successful control or eradication efforts. While eDNA methods are promising, most require time-consuming sample processing and DNA extraction steps, all of which require a laboratory. In this study, a field-based alternative to traditional eDNA measurement methods was developed which included: i) a rapid sampling technique which uses a 35 µm mesh to recover and concentrate larger cells, ii) an extensive volunteer network to allow collection of samples from throughout Michigan, iii) isothermal amplification of eDNA using a field-deployable device (Gene-Z) and microfluidic chips, and iv) reporting of results through an Android- based application termed iSAW (invasive species appearance warning). In all, over 200 samples were collected in the 2014 sampling season from inland lakes in the state of Michigan and analyzed for a number of current and high-risk potential invasive species including Dreissena polymorpha and bugensis, Limnoperna fortunei, Cercopagis pengoi, Dikerogammarus villosus, Bythotrephes longimanus, Daphnia cristata, Channa argus, Neogobius melanostomus, Orconectes rusticus and Petromyzon marinus. Results from this study are viewable in the iSAW application as well as on Google Maps. To our knowledge, this study represents the first comprehensive invasive species monitoring program of its kind, which includes the developed sampling approach, use of a network of sampling volunteers and novel technology.

This work was supported in part by United States EPA Great Lakes Restoration Initiative (GLRI) grant number GL-00E01127-0


ENE-05  Temporal Patterns Of Chloride And Sodium In A Suburban Watershed During Snow Melting

Authors: Eunsang Lee; Huiyun Wu; David T. Long; Thomas C. Voice

Abstract: It has been shown that levels of sodium and chloride in surface waters have been increasing over several decades due to the use of deicers on roadways. It has been challenging to study the dynamic behavior of these ions because it is difficult to separate the effects of release patterns and pathways from interactions with soils. For example, it is well known that sodium can be retained and later released during transport through soils by cation exchange. In this study, we collected samples from the Red Cedar River one to two times per day over the course of a spring snow-melt event and measured sodium and chloride concentrations. We also collected occasional samples from snowpack and puddles on the roadside in order to evaluate locations that could serve as significant sources. The results showed that: 1) samples from the river showed a peak in sodium and chloride concentrations early in the period of snow-melt, suggesting a first-flush phenomenon, 2) applied deicer was not representative of pure NaCl, having a ratio of about 0.5 rather than 1, and 3) Na/Cl ratios from suspected source locations decreased over the snow-melt period, while ratios in the stream were stable and similar to those in source locations over the first-flush period, but increased afterwards. This suggests that the first-flush of these ions originates largely from residual deicing chemicals proximate to the stream, whereas longer-term behavior is likely linked to longer transport pathways with significant interactions between ions and soils.

This work was supported in part by U.S.G.S. Michigan RASA project, MSU Water Initiative and the National Institute of Health


ENE-06  Bacteriophage Diversity And Abundance In Activated Sludge And Potential Applications

Authors: Camille McCall; Mariya Munir; Terence Marsh; Irene Xagoraraki

Abstract: Bacteriophages have been identified as the largest viral group in the environment, and continue to predominate in wastewater treatment plants over any natural aquatic system. Tailed phages constitute the largest order of bacteriophages, Caudovirales, which is composed of three families, Siphoviridae, Myoviridae, and Podoviridae. Polyhedral, filamentous, and pleomorphic type phages represent a smaller percentage of bacterial viruses. Bacteriophages play a large role in the structure of the microbial biosphere and have thus proven themselves major influencers in larger systems, such as water and wastewater treatment systems, food, and health industries. Since the discovery of bacteriophages in the early 1900’s over 5,000 phages have been recognized by the International Center of Taxonomy of Viruses (ICTV). A large concentration of bacteriophages in wastewater treatment systems is found in the activated sludge process due to its favorable environment. This study utilizes metagenomic analysis to compare the diversity and abundance of phages in the activated sludge of two different wastewater treatment processes. It was reported that Caudovirales comprises 92% of affiliated sequences with Siphoviridae being the most abundant family of bacteriophages followed by Podoviridae, and Myoviridae. Pseudomonas phages were also identified as the most abundant bacteriophage species type in both samples, which give insight into the bacterial composition of the activated sludge. This study offers the abundance and diversity of phages in activated sludge, which can be utilized to explore possible applications of these phages in food, plant disease control, wastewater and water treatment, and medicine. Results show activated sludge as an ideal environment for isolating virulent bacteriophages useful for phage applications.


ENE-07  Screening For Potential Viral Pathogens In Wastewater Effluent And Activated Sludge From An MBR And A Conventional Treatment Utility Using Metagenomics Analysis

Authors: Mariya Munir; Evan O’Brien; Terence Marsh; Irene Xagoraraki

Abstract: Despite recent rapid advancements in water and wastewater treatment technologies, waterborne pathogens still remain as one of the major environmental threats to human health. Monitoring of all pathogens with conventional methods is not feasible due to time and cost constraints. In this paper, viral diversity of two wastewater treatment plant effluents, one a conventional activated sludge (CAS) facility and one a membrane bioreactor (MBR) facility, are investigated using metagenomics. Diversity analysis does not provide quantitative data on pathogen loads or infectivity but it provides a list of potentially pathogenic viruses that need to be considered in more detail. The most abundant potential human viral pathogen observed in our study belongs to taxonomic order Herpesvirales. Other potentially pathogenic viruses detected in this study include Poxviridae, Adenoviridae, and Coronaviridae. Metagenomic analyses in this study also revealed that a large proportion of sequences could not be assigned to taxonomic affiliations even at the phylum/class levels and thus are most likely to be derived from novel, uncharacterized microbes. This paper provides guidance on which viral pathogens to monitor in the effluents of WWTPs especially in case of wastewater reuse.


ENE-08  Vinyl Chloride Assimilators DNA Biomarker Design For Future Field Applications

Authors: Fernanda Paes; Timothy Mattes; Alison Cupples

Abstract: Vinyl chloride (VC) is a common groundwater contaminant and a known carcinogen. The use of microorganisms to treat contaminated sites (bioremediation) is a technology with great potential and lower costs compared to other remediation approaches. Identifying which microorganisms are key degraders of the targeted chemical is important if bioremediation is desired. Here, stable isotope probing (SIP) was combined with Illumina sequencing and used to identify microorganisms able to uptake carbon from the 13C- labeled VC in a mixed culture. Microcosms were constructed using mineral media, micronutrients, mixed culture and amended with 13C-labeled or 12C-unlabeled VC. On days 3 and 7, 13C-labeled and 12C-unlabeled VC microcosm’s replicates were sacrificed for DNA extraction. DNA was ultracentrifuged, fractioned, and the fractions of greater buoyant density (heavy fractions) were subject to Illumina sequencing. Specific primers (biomarkers) were designed for the identified phylotypes using Primer-BLAST. The new primers were tested for specificity by Sanger sequencing and used in qPCR assays across the buoyant density gradient to confirm label uptake by these phylotypes. Four phylotypes were identified as VC-assimilators: Nocardioides, Sediminibacterium, Aquabacterium and Variovorax. Both previously identified VC-assimilating (Nocardioides) as well as novel genera, were responsible for VC-carbon uptake (or VC degradation products). The designed biomarkers were successfully validated to the tested culture. Using biomolecular technologies to assess VC-assimilators from mixed cultures is important because there is likely a greater diversity of VC-assimilators in the environment than is currently represented in pure culture. The biomarkers designed in this work could be helpful to assess these phylotypes in future field studies.

This work was supported in part by NSF; SERDP


ENE-09  Biological Degradation Of Carbamazepine In Anaerobic Soils

Authors: Jean-Rene Thelusmond; Alison M. Cupples

Abstract: Carbamazepine (CBZ) is an anti-epileptic drug encountered in wastewater effluent and surface water due to its refractory behavior to biodegradation during wastewater treatment. Introduced into agricultural soils primarily via irrigation with reclaimed wastewater and biosolids application, CBZ has been recognized to accumulate in soils. This study aims at investigating carbamazepine biodegradability in soils subjected to anaerobic conditions and determining the responsible microorganisms. Prior to establishing the microcosms and measuring degradation, methods were developed in order to extract spiked CBZ from soils by using the modified QuECHERS which stands for “quick, easy, cheap, effective, rugged, and safe.” The extraction procedure consisted of adding acetonitrile to soil-CBZ-deionized water mixture before slowly and continuously adding the QuECHERS buffer. The extraction was then followed by the purification of the extracts by solid phase extraction (SPE) using a strong anion exchange cartridge (SAX) in tandem with a polymeric cartridge (Oasis HLB) before the samples were analyzed by liquid chromatography electrospray tandem mass spectrometry (LC-ESI-MS/MS) performed on a 3200 QTRAP. Ascentis ® Express C18 column was used, and the mobile phases were (A) 1% formic acid in MilliQ water and (B) acetonitrile. The flow rate was 0.3 ml/min and the injection volume 10 µl. The average recovery for CBZ in soil was 103.78±1.42 for the tested concentration (500 ng g-1). On-going research focuses on determining CBZ biodegradation rates in three agricultural soils. In addition, the effect of CBZ on soil microbial communities will be investigated with high throughput sequencing.

This work was supported in part by U.S Department of Agriculture (USDA)


ENE-10  Utilizing Fermention Byproducts To Enhance Hydrogen Production Using Spinel Photocatalysts

Authors: Xiaoyu Wang; Susan Masten; Simon Davies

Abstract: Recent research in fuel cell technology using hydrogen to store solar energy in a photovoltaic array still cannot meet the demand for electricity, so semiconductor photocatalytically mediated water splitting has been widely investigated. However, with a narrow band absorbing visible light, a semiconductor is often not sufficiently energetic to oxidize water. Using organic wastes or wastewater containing fermented biomass with spinel photocatalysts is supposed to solve the problem and improve hydrogen production. The objective of this study is to test the ability of several compounds commonly found in fermented biomass to produce hydrogen using spinel photocatalysts and sunlight, select a limited number of scavengers effective for hydrogen production, and clarify the factors that affect the efficacy of hole scavengers enhances hydrogen production for obtaining higher solar energy storage efficiency in photocatalyst system in the future. Therefore, we will produce heterojunction photocatalyst and examine the extent to which the intermediates formed during its photodegradation enhance hydrogen production. In the system, selected organic compounds in the fermented biomass are expected to improve photocatalytic hydrogen production as sacrificial hole scavengers by orders of magnitude compared to that in the absence of scavengers. This proposed process would oxidize the organic material in the waste and at the same time produce energy.

This work was supported in part by Discretionary Funding Initiative MSU Internal Funding; MSU College of Engineering; Department of Civil and Environmental Engineering


ENE-11  Wind Fetch Length Impacts On Coastal Bacteria Loading In Southwestern Lake Michigan

Authors: Chelsea Weiskerger; Meredith Nevers; Phanikumar Mantha; Richard Whitman

Abstract: The bacterial content observed at beaches along Lake Michigan’s southwestern shore varies on both temporal and spatial scales. Previous research indicates that these variations are caused by both point and nonpoint sources of contamination, but it is reasonable to think that physical and geological conditions also have effects on the bacterial loading at these beaches. In this study, daily wind fetch length was characterized at twenty beach sampling sites in Chicago, Illinois, over the course of three field seasons. These daily wind fetch lengths were compared to daily counts of Escherichia coli (E. coli) bacteria at each location, to determine the extent of influence that wind fetch length has on bacterial loading and risk of beach closure due to high E. coli counts. While the resulting data do not suggest a direct influence of wind fetch length on raw bacterial counts, a Spearman Correlation test indicates that among the beaches, wind fetch length is significantly correlated (α = 0.01) with high risk of beach advisory or closure. The data show that as wind fetch length decreases, probability of advisory or closure due to contamination increases. This may be due to the prevalence of embayments and breakwaters along Chicago’s shore. Prevailing winds are often out of the north and run along the shoreline in Chicago. These embayments and breakwaters decrease wind fetch length while increasing the capacity for the shore to trap bacteria over time, thus leading to high contamination rates and risk of advisory or closure.

This work was supported in part by United States Geological Survey, contract G13PX01435


ENE-12  Temporal Patterns Of DOC And K+ Concentrations In Salted Urban Watersheds

Authors: Huiyun Wu; Eunsang Lee; Thomas Voice; David Long

Abstract: First flush refers to a phenomenon where the concentrations of chemical species increase dramatically before the peak of flow during a runoff event. In this study, we measured the temporal concentration patterns of dissolved organic carbon (DOC), potassium (K+) and specific ultraviolet absorbance (SUVA) at 254 nm, and evaluated the relationship of these parameters to streamflow rate, soil temperature, precipitation and snow-pack depth for a spring snow-melt event in 2014 and 2015.  In 2014, it was found that DOC exhibited first-flush behavior, with a series of concentration peaks that decreased with subsequent precipitation/melting events.  Average DOC concentrations increased over the melting period, nearly doubling.  The trends for K+ were similar.  Preliminary analysis of 2015 data shows that SUVA 254 increases over the first-flush peak.  Since DOC originates largely degradation of plant material on the landscape and SUVA measurements provide insight into the extent of degradation, the results may inform our understanding of sources and transport pathways.  This will be explored in further analysis of the data collected.

This work was supported in part by DEQ