Case Study on San Jacinto Basin This is our generic title slide. Not sure what to place for the title so its left blank atm. Case Study on San Jacinto Basin Rupa Iyer Ph.D. Founding Director, Biotechnology Programs Director, Center for Life Sciences Technology University of Houston

Biotechnology Medicine Food Pharmaceutical Agriculture Environmental • improving human, animal, and plant health; • increasing the availability of nutritious food products • reducing our reliance on chemical pesticides Agriculture Environmental

Environmental Sampling Remediation of OP Compounds UH Interdisciplinary Research Based Laboratory Curriculum Discovery R & D Production Applications Environmental Sampling Remediation of OP Compounds Biomanufacturing Module I Sample Collection & Screening Module II Cloning & Transformation of OP Genes Module III Upstream & Downstream Processing of OP Hydrolase Bioremediation Biosecurity Energy Health Entrepreneurship Infused 20 years of research in biotechnology laboratory curriculum Pesticide degrading bacteria as a model to demonstrate the life cycle of a biotech product Modular format , ease of integration in a variety of courses Seed Funding NSF*, TWC Model for interdisciplinary Education

Environmental Sampling Research & Education No comments. A hands-on research experience for undergraduates with real world applications

Database of Environmental Contamination Activity – Research Collaboration Network A hands-on research experience for undergraduates with real world applications engages students in an authentic, ongoing research activity to populate a database of the geographic distribution of environmental contamination Electronic archive of experimental data – *Students contributing and mining their own research data to enhance STEM skills

A Database of Environmental Activity The ESRM database around the Houston-metropolitan area. AN old picture, doesn’t appear to include SJR sites. Discuss in context of the ESRM before our extended focus on the SJR.

Integration of Education and Research - Outcomes 60+ bacterial strains Map of location and distribution pesticide degradation activity 2 distinct organophosphate (OP) degradation genes *Genetic Map of 8 bacteria Check font and formatting for consistency.

Platform for Education and Research Collaboration Students: Hands-on STEM activities and Research Experiences Faculty: Implications and applications in biotechnology Government and Community : Location, monitoring, and remediation efforts Commercial: Diverse collection of bacteria available for purchase- bioremediation, enhanced oil recovery, antibiotic resistance Consider switching this with the slide below.

Platform for Education and Research Collaboration Local UH BTEC Prairie View A&M Alvin Community College National BYU-Hawaii Louisiana Tech RIT SUSU Global St. Xavier's (India) Morocco Fulbright Specialist (5 years) No comments. Addressing two social Issues : STEM & the Environment

San Jacinto Waste Pits – Case study Location – Two pits: Consists of two pits under 1-10 overpass as it crosses the San Jacinto River 14 acres total in size Pits partially submerged releasing contamianants into and across the SJR. Location Consists of two pits under 1-10 overpass as it crosses the San Jacinto River Purpose Champion Paper Mill dumping ground for paper mill waste in the 1960’s Contaminants Polychlorinated organics Dibenzo-p-dioxins Dibenzofurans Biphenyls

San Jacinto Waste Pits - History Location Consists of two pits under 1-10 overpass as it crosses the San Jacinto River 14 acres total in size Purpose Champion Paper Mill dumping ground for paper mill waste in the 1960’s Contaminants Polychlorinated organics Dibenzo-p-dioxins Dibenzofurans Biphenyls Location – Two pitsConsists of two pits under 1-10 overpass as it crosses the San Jacinto River 14 acres total in size

San Jacinto Waste Pits - History 1990 First dioxin based consumption advisory for fish and shellfish for HSC and Galveston Bay 2005 San Jacinto Waste pits superfund site was rediscovered. 2011 Waste Management temporary cap to minimize release Location – Two pitsConsists of two pits under 1-10 overpass as it crosses the San Jacinto River 14 acres total in size

San Jacinto Waste Pits - Legacy Areial Map; Red is the superfund and immediate surrounding area. Yellow, census tracks representing cancer cluster. Official report does not exist. TDSHS Report: Noted increased incidences in cancer in various eastern Harris county census tracts. Three of those tracts overlap on top of the Superfund and extend downstream towards Galveston Bay. Widespread contamination of sediment, surface water, soil, and aquatic wildlife downstream of waste pits San Jacinto River Houston Ship Channel Buffalo Bayou Galveston Bay Health risks Texas Department of State Health Services Cancer Report

Map of Dioxin Monitoring Stations Shows the location of dioxin monitoring stations across the SJR and down into Galveston Bay. Pins are color coded based on the area not concentration. Colors do not correspond to TEQs on next slide. Yellow – Buffalo Bayou Pink – Bear Lake Light blue surrounded by red outline – Superfund Blue – Lost Lake Light pink – Burnet Bay Green – Scott Bay Red – Upper SJR Bay Light blue – Black duck Bay Dark blue – Tabbs bay Purple – Mouth of SJR Orange – Galveston Bay

San Jacinto River Collection - Project Rationale Detect Pollution Bacteria as biomarkers Remediation In situ bioremediation No comments. Develop a distribution map of xenobiotic-degrading activity across SJR Dioxins Furans Biphenyls Petroleum Hydrocarbons Pesticides Other Organic Waste Biomarkers Degradation Activity Development of bacterial biosensorsAdd to a collection of xenobiotic degrading bacteria that can be utilized for environmental cleanup of these pollutants SJR Superfunds

Project Goals (1) Collect environmental samples from San Jacinto River Soil and sediment Project Goals Samples were taken at multiple locations along the San Jacinto river in Fall 2016 – Spring 2017. - Samples are color coded as follows: Blue – No isolate obtained from sample. Yellow – Positive isolate (paraoxon). Purple – Positive isolate (methyl parathion). Green – Positive isolate (dioxin/dibenzofuran). Orange – Positive isolate (1,4 dioxane). Brown – Positive isolate (biphenyl). Light green – Positive (paraoxon/dioxin) Blue – No isolate obtained from sample. Yellow – Positive (paraoxon). Purple – Positive (methyl parathion). Green – Positive (dioxin). Brown – Positive (biphenyl). Light green – Positive (paraoxon/dioxin)

Project Goals (2) Screen collected environmental samples for dioxin and other xenobiotic degrading bacteria Ops on the top. Central phosphorus atom. Dioxins Furans and Biphenyls. Very similar structures. Many degradation genes exhibit cross activity especially in the presence of a preferred co-substrate. 1,4 Dioxane and a similar compound. Tetrahydrofuran monooxygenase is the enzyme that targets 1,4 dioxane.

Preliminary Project Results: Identification of SJR Bacterial Isolates Genera + species screened out on specific substrates from areas along the SJR. Updated Table. This is in picture form. See supplementary slide for editable table.

Project Goals (3) Identify dioxin degradation biomarker genes and functional degradation pathways Whole genome sequencing Data analysis Characterization Genomics – DNA sequencing (Represents the biological potential of your organism(s)) Transciptnomics – RNA sequencing, Microarray (Represents gene expression of different biological activities) Proteomics – Protein modeling (Represents actual biological activity taking place inside of organism) Metablomics – Degradation kinetics (The end result of biological activity in your organism) Together this data allows you to determine what your organism can do and how you can improve upon it. Potential Function Activity DNA RNA Protein Metabolites

Project Goals (4) Map spatial-genomic distribution of dioxin degrading activity Phylogeographic analyses (5) Engineer bacteria for improved degradation of dioxin compounds Last goal slide. Analysis of degradation activity and directed improvement to degradation enzymes. Genomic insertion, deletion, or editing Mutagenesis CRISPR-Cas

Preliminary Project Results: General Trends Many isolated bacteria so far are noted opportunistic pathogens or known pathogens Polluted environments are hotspots for later transfer SJR may be a breeding ground for antibiotic resistance Data mirrors previous results taken over 8 years of undergraduate environmental sampling in agricultural soils for organophosphate degradation High incidence of opportunistic pathogens as OP degraders Evidence of lateral transfer of antibiotic resistance genes observed in a subset of collected isolates Manure was a likely source of resistance genes in these instances No comments.

Environmental Ideals Not too sure about slide title. Meant to depict a pristine river devoid of pollution.