HOUSEHOLD HAZARDOUS WASTE PROGRAM: POPULATION DEMOGRAPHICS

Master of Public HealthPublic Health Interdepartmental ProgramMelinda WilkersonMy master’s project and field experience provided me with a wide variety of opportunities for personal growth, as well as acquainted me with the different aspects of the field of public health. My capstone project developed a method by which the Johnson county Household Hazardous Waste site was able to consolidate their data in a way to quickly analyze the results from the 92,402 appointments they have developed from the past 12 years and will continue to add data. Principally my project and field experience focused on organization of data and analysis of the areas we serve, and potential demographics or areas of the county the site might not be serving sufficiently. I was able to visualize the data through the use of a geographic information system. This program allows public health workers to visualize, question, analyze, and interpret data to understand relationships, patterns, and trends. This program was essential to understanding what is happening and what might happen in a geographic space. The household hazardous waste site is a county funded facility that is critical to the health and safety of not only the environment by keeping hazardous waste out of the city dumps, but also gives people a place to dispose of their environmentally hazardous chemicals in a safe way. Lastly my project was accepted by the North American Hazardous Materials Management Association (NAHMMA) to be presented, where my preceptor Julie Davis will present the data and PowerPoint I put together for her. In this report, I will discuss the project and field experienc

Molecular Mechanisms Driving Switch Behavior in Xylem Cell Differentiation.

Plant xylem cells conduct water and mineral nutrients. Although most plant cells are totipotent, xylem cells are unusual and undergo terminal differentiation. Many genes regulating this process are well characterized, including the Vascular-related NAC Domain 7 (VND7), MYB46, and MYB83 transcription factors, which are proposed to act in interconnected feedforward loops (FFLs). Less is known regarding the molecular mechanisms underlying the terminal transition to xylem cell differentiation. Here, we generate whole-root and single-cell data, which demonstrate that VND7 initiates sharp switching of root cells to xylem cell identity. Based on these data, we identified 4 candidate VND7 downstream target genes capable of generating this switch. Although MYB46 responds to VND7 induction, it is not among these targets. This system provides an important model to study the emergent properties that may give rise to totipotency relative to terminal differentiation and reveals xylem cell subtypes

Linkage of Marine Bacterial Polyunsaturated Fatty Acid and Long-Chain Hydrocarbon Biosynthesis

Various marine gamma-proteobacteria produce omega-3 polyunsaturated fatty acids, such as eicosapentaenoic acid (20:5, EPA) and docosahexaenoic acid (22:6, DHA), which are incorporated into membrane phospholipids. Five genes, designated pfaABCDE, encode the polyketide/fatty acid synthase necessary for production of these long-chain fatty acids. In addition to de novo biosynthesis of EPA and DHA, the “Pfa synthase” is also involved with production of a long-chain polyunsaturated hydrocarbon product (31:9, PUHC) in conjunction with the oleABCD hydrocarbon biosynthesis pathway. In this work, we demonstrate that OleA mediates the linkage between these two pathways in vivo. Co-expression of pfaA-E along with oleA from Shewanella pealeana in Escherichia coli yielded the expected product, a 31:8 ketone along with a dramatic ∼10-fold reduction in EPA content. The decrease in EPA content was independent of 31:8 ketone production as co-expression of an OleA active site mutant also led to identical decreases in EPA content. We also demonstrate that a gene linked with either pfa and/or ole operons in diverse bacterial lineages, herein designated pfaT, plays a role in maintaining optimal production of Pfa synthase derived products in Photobacterium and Shewanella species

Widespread Occurrence of Secondary Lipid Biosynthesis Potential in Microbial Lineages

Bacterial production of long-chain omega-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), is constrained to a narrow subset of marine γ-proteobacteria. The genes responsible for de novo bacterial PUFA biosynthesis, designated pfaEABCD, encode large, multi-domain protein complexes akin to type I iterative fatty acid and polyketide synthases, herein referred to as “Pfa synthases”. In addition to the archetypal Pfa synthase gene products from marine bacteria, we have identified homologous type I FAS/PKS gene clusters in diverse microbial lineages spanning 45 genera representing 10 phyla, presumed to be involved in long-chain fatty acid biosynthesis. In total, 20 distinct types of gene clusters were identified. Collectively, we propose the designation of “secondary lipids” to describe these biosynthetic pathways and products, a proposition consistent with the “secondary metabolite” vernacular. Phylogenomic analysis reveals a high degree of functional conservation within distinct biosynthetic pathways. Incongruence between secondary lipid synthase functional clades and taxonomic group membership combined with the lack of orthologous gene clusters in closely related strains suggests horizontal gene transfer has contributed to the dissemination of specialized lipid biosynthetic activities across disparate microbial lineages

fabH deletion increases DHA productionin Escherichia coli expressing Pfa genes

Background: Some marine bacteria, such as Moritella marina, produce the nutraceutical docosahexaenoic acid (DHA) thanks to a specific enzymatic complex called Pfa synthase. Escherichia coli heterologously expressing the pfa gene cluster from M. marina also produces DHA. The aim of this study was to find genetic or metabolic conditions to increase DHA production in E. coli. Results: First, we analysed the effect of the antibiotic cerulenin, showing that DHA production increased twofold. Then, we tested a series of single gene knockout mutations affecting fatty acid biosynthesis, in order to optimize the synthesis of DHA. The most effective mutant, fabH, showed a threefold increase compared to wild type strain. The combination of cerulenin inhibition and fabH deletion rendered a 6.5-fold improvement compared to control strain. Both strategies seem to have the same mechanism of action, in which fatty acid synthesis via the canonical pathway (fab pathway) is affected in its first catalytic step, which allows the substrates to be used by the heterologous pathway to synthesize DHA. Conclusions: DHA-producing E. coli strain that carries a fabH gene deletion boosts DHA production by tuning down the competing canonical biosynthesis pathway. Our approach can be used for optimization of DHA production in different organisms.Funding: The work in the FdlC and GM laboratories was financed by the Spanish Ministry of Economy, Industry and Competitiveness Grant BFU2014-55534-C2

Secondary metabolite gene expression and interplay of bacterial functions in a tropical freshwater cyanobacterial bloom

Cyanobacterial harmful algal blooms (cyanoHABs) appear to be increasing in frequency on a global scale. The Cyanobacteria in blooms can produce toxic secondary metabolites that make freshwater dangerous for drinking and recreation. To characterize microbial activities in a cyanoHAB, transcripts from a eutrophic freshwater reservoir in Singapore were sequenced for six samples collected over one day-night period. Transcripts from the Cyanobacterium Microcystis dominated all samples and were accompanied by at least 533 genera primarily from the Cyanobacteria, Proteobacteria, Bacteroidetes and Actinobacteria. Within the Microcystis population, abundant transcripts were from genes for buoyancy, photosynthesis and synthesis of the toxin microviridin, suggesting that these are necessary for competitive dominance in the Reservoir. During the day, Microcystis transcripts were enriched in photosynthesis and energy metabolism while at night enriched pathways included DNA replication and repair and toxin biosynthesis. Microcystis was the dominant source of transcripts from polyketide and non-ribosomal peptide synthase (PKS and NRPS, respectively) gene clusters. Unexpectedly, expression of all PKS/NRPS gene clusters, including for the toxins microcystin and aeruginosin, occurred throughout the day-night cycle. The most highly expressed PKS/NRPS gene cluster from Microcystis is not associated with any known product. The four most abundant phyla in the reservoir were enriched in different functions, including photosynthesis (Cyanobacteria), breakdown of complex organic molecules (Proteobacteria), glycan metabolism (Bacteroidetes) and breakdown of plant carbohydrates, such as cellobiose (Actinobacteria). These results provide the first estimate of secondary metabolite gene expression, functional partitioning and functional interplay in a freshwater cyanoHAB.Singapore. National Research Foundation (Singapore MIT Alliance for Research and Technology (SMART), Center for Environmental Sensing and Modeling (CENSAM) research program)National Science Foundation (U.S.) (Postdoctoral Research Fellowship in Biology, Grant No. DBI-1202865)National Institute of Environmental Health Sciences (NIEHS Grant P30-ES002109 to the MIT Center for Environmental Health Sciences)MIT International Science and Technology Initiatives (MISTI-Hayashi fund

The Natural Product Domain Seeker NaPDoS: A Phylogeny Based Bioinformatic Tool to Classify Secondary Metabolite Gene Diversity

New bioinformatic tools are needed to analyze the growing volume of DNA sequence data. This is especially true in the case of secondary metabolite biosynthesis, where the highly repetitive nature of the associated genes creates major challenges for accurate sequence assembly and analysis. Here we introduce the web tool Natural Product Domain Seeker (NaPDoS), which provides an automated method to assess the secondary metabolite biosynthetic gene diversity and novelty of strains or environments. NaPDoS analyses are based on the phylogenetic relationships of sequence tags derived from polyketide synthase (PKS) and non-ribosomal peptide synthetase (NRPS) genes, respectively. The sequence tags correspond to PKS-derived ketosynthase domains and NRPS-derived condensation domains and are compared to an internal database of experimentally characterized biosynthetic genes. NaPDoS provides a rapid mechanism to extract and classify ketosynthase and condensation domains from PCR products, genomes, and metagenomic datasets. Close database matches provide a mechanism to infer the generalized structures of secondary metabolites while new phylogenetic lineages provide targets for the discovery of new enzyme architectures or mechanisms of secondary metabolite assembly. Here we outline the main features of NaPDoS and test it on four draft genome sequences and two metagenomic datasets. The results provide a rapid method to assess secondary metabolite biosynthetic gene diversity and richness in organisms or environments and a mechanism to identify genes that may be associated with uncharacterized biochemistry

The role of plant growth promoting bacteria on arsenic removal: a review of existing perspectives

Phytobial remediation is an innovative tool that uses plants and microbes to mitigate Arsenic (As) contamination of the environment. Recently, plant growth-promoting bacteria (PGPB) that assists phytoremediation has been highly touted for both improving plant metal tolerance and promoting plant growth while achieving the goal of large-scale removal of As. This review focuses on the PGPB characteristics influencing plants and the mechanisms in which they function to overcome/lessen As-induced adversities. Several recent examples of mechanisms responsible for increasing the availability of As to plants and coping with As stresses facilitated by PGPB will be reviewed. Although drawbacks to phytoremediation have been reported, encouraging results have been developed with regular monitoring. Introducing PGPB-assisted phytoremediation of As in a field requires an assessment of the environmental effects of PGPB, especially with respect to the impacts on indigenous bacteria

The ecological role of ponds in a changing world

The fifth conference of the European Pond Conservation Network (Luxembourg, June 2012) brought together researchers, environmental managers, and other stakeholders with the aim to share stateof-the-art knowledge on the ecology, management, and conservation of ponds in the context of the many challenges facing the wider water environment. Although well-known ecological patterns apply to most ponds in Europe and elsewhere, recent data highlight that part of the environmental variables governing pond biodiversity remain specific to climatic/ biogeographic regions and to elevation ranges, suggesting that, in addition to common practice, management plans should include range-specific measures. Beyond the contribution of individual ponds to the aquatic and terrestrial life, connected networks of ponds are vital in the provision of new climate space as a response to global climate change, by allowing the observed northward and/or upward movements of species. In terms of services, ponds offer sustainable solutions to key issues of water management and climate change such as nutrient retention, rainfall interception, or carbon sequestration. While the ecological role of ponds is now well established, authoritative research-based advice remains needed to inform future direction in the conservation of small water bodies and to further bridge the gap between science and practice

The ecological role of ponds in a changing world

The fifth conference of the European Pond Conservation Network (Luxembourg, June 2012) brought together researchers, environmental managers, and other stakeholders with the aim to share stateof-the-art knowledge on the ecology, management, and conservation of ponds in the context of the many challenges facing the wider water environment. Although well-known ecological patterns apply to most ponds in Europe and elsewhere, recent data highlight that part of the environmental variables governing pond biodiversity remain specific to climatic/ biogeographic regions and to elevation ranges, suggesting that, in addition to common practice, management plans should include range-specific measures. Beyond the contribution of individual ponds to the aquatic and terrestrial life, connected networks of ponds are vital in the provision of new climate space as a response to global climate change, by allowing the observed northward and/or upward movements of species. In terms of services, ponds offer sustainable solutions to key issues of water management and climate change such as nutrient retention, rainfall interception, or carbon sequestration. While the ecological role of ponds is now well established, authoritative research-based advice remains needed to inform future direction in the conservation of small water bodies and to further bridge the gap between science and practice