Bacterial Source Tracking Study. BST Assessment of Fecal Pollution Sources Impacting the Trinity River Floodway
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1 Bacterial Source Tracking Study BST Assessment of Fecal Pollution Sources Impacting the Trinity River Floodway
2 Objective Ten sets of water samples were collected from each of 8 sites over an 10 week period in an attempt to identify sources contributing to bacterial loading in the Trinity River Floodway.? 2
3 PCR Sample Analysis (Library Independent) Method 2 Methods for E. coli Source Identification The University of Texas Health Science Center at Houston (UTHealth) School of Public Health in El Paso DNA RiboPrint (Library Dependent) Used during first 2 weeks of study Used for last 8 weeks of study 3B Acetaminophen Caffeine Diltiazem Ketamine Sulfamethoxazole Baylor University Chemical Tracers Supplemental Support Data Collected Benzoylecogine Carbamazepine Diphenhydramine Sucralose Trimethoprim Temperature ph Dissolved Oxygen Specific Conductance Turbidity TDS Pace Analytical Water Quality NH3 NOx TKN TP OPO4 Chloride E. coli (Idexx) 3
4 Sampling Sites West 199 (Henderson) (WF1) West Confluence (WFC) Clear Kayak Creek Chute #4 (CFK) Clear Trinity Park (CFT) Zoo Colonial (ZCC) Zoo McPherson (ZCM) Clear Rogers Rd (CFR) Clear Bryant Irving (CFB) 4
5 TRWD BST of qpcr Data, Weeks1-2 The first two sets of water samples were analyzed for the qpcr markers: (1) Bacteroidales GenBac3 (general/non-specific) a) HF183/BacR287 (human) b) BacCow ruminant (and other animals) c) Helicobacter GFD (bird) 5
6 qpcr Results Weeks 1&2 Log Total Bacteroidales GenBac3 Marker Figure 2: A closer look at the Bacteroidales GenBac3 Marker, which gives an estimate of overall fecal pollution levels at each of 8 sites in the Trinity. 6
7 qpcr Results Weeks 1&2 Log Human HF183 Marker The Human HF183 Marker suggests that the site Zoo Creek at McPherson (ZCM) is significantly impacted by human fecal pollution; to a lesser extent, the sites Clear Fork at the Kayak Chute (CFK) and Zoo Creek at Colonial (ZCC) also appear to be impacted by human fecal pollution 7
8 qpcr Results Weeks 1&2 Log GFD Bird Marker
9 qpcr Results Weeks 1&2 Log Ruminant-Animal BacCow Marker
10 qpcr Results Weeks 1&2 TRWD BST qpcr Batches 1 and 2 Log Total Bacteroidales GenBac3 Marker Log GFD Bird Marker Log Human HF183 Marker Log Ruminant-Animal BacCow Marker Log Marker Copy Number Site 10
11 Log Total Bacteroidales vs Log E. coli 11
12 DNA Riboprinting/Library Dependent Analysis E. coli samples cultured on modified mtec plates Up to 10 isolates collected from each cultured plate (some E. coli counts too low to collect 10 isolates) 615 isolates collected in total from 64 samples. ERIC-RP fingerprinting done and identified using the Texas E. coli BST Library (ver. 5-15) As with any BST method, percentages (or marker quantification) do not necessarily equate to percentage of fecal pollution loading--rather, results allow the identification and ranking of major fecal pollution sources. 12
13 Human Sewage Septic 4-Way Library Dependent Classifications Domestic Animals Cat Dog Duck Turtle Cattle Chicken Duck Goose Guinea Fowl Turkey Donkey Goat Hog Horse Llama Pig Rabbit Sheep Avian Wildlife Blackbird Buzzard Blue jay Brown headed cowbird Crow Double crested cormorant Dove Duck Egret Goose Grackle Heron Killdeer Mockingbird Pelican Pigeon Roadrunner Gull Sparrow Starling Swallow Tern Turkey Vulture Non-Avian Wildlife Armadillo Beaver Bobcat Coyote Deer Feral hog Fox Javelina Mouse Opossum Porcupine Prairie dog Rabbit Raccoon Rat Skunk Squirrel Wolf
14 Isolate Data E. Coli Isolate MPN/100 ml Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr Bryant Irving Rogers Rd Zoo Creek McPherson Zoo Creek Colonial City Dam Clear Fork Kayak West Fork 199 West Fork Confluence Human Domestic Avian Wildlife Non-Avian Wildlife Unidentified 14
15 Clear Bryant Irving Geometric Mean = 65 CFU/100 ml (n=68 isolates; 8 samples) 15
16 Clear Rogers Rd. Geometric Mean = 97 CFU/100 ml (n=80 isolates; 8 samples) 16
17 Zoo McPherson Geometric Mean = 2635 CFU/100 ml (n=80 isolates; 8 samples)
18 Zoo Colonial Geometric Mean = 786 CFU/100 ml (n=80 isolates; 8 samples)
19 Clear Trinity Park Geometric Mean = 314 CFU/100 ml (n=80 isolates; 8 samples) 19
20 Geometric Mean = 122 CFU/100 ml (n=80 isolates; 8 samples) Clear Kayak Chute #4
21 West 199 (Henderson) Geometric Mean = 99 CFU/100 ml (n=77 isolates; 8 samples)
22 West Confluence Geometric Mean = 77 CFU/100 ml (n=70 isolates; 8 samples) 22
23 Four-way Split of Source Classes Figure 8: Identification of E. coli water isolates using a four-way split of source classes. Results are normalized to percent of isolates per sampling date (67 to 80 isolates per date; all sites combined). Note: samples taken 3-Apr and 18-Apr were after rain events. 23
24 1800 Isolate Data E. Coli Isolate: Human MPN/100 ml Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr 13-Feb 27-Feb 6-Mar 20-Mar 27-Mar 3-Apr 10-Apr 18-Apr Bryant Irving Rogers Rd Zoo Creek McPherson Zoo Creek Colonial City Dam Clear Fork Kayak West Fork 199 West Fork Confluence 24
25 Load Data E.coli Load in the Trinity 3E E+13 2E+13 MPN/Day 1.5E+13 1E+13 5E /10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 Bryant Irving Rogers Rd Zoo Creek McPherson Zoo Creek Colonial City Dam Clear Fork Kayak West Fork 199 West Fork Confluence Human Domestic Avian Wildlife Non-Avian Wildlife Unidentified 25
26 Load Data Human E.coli Load in the Trinity 3E E+12 2E+12 MPN/Day 1.5E+12 1E+12 5E /10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 4/10/2017 Bryant Irving Rogers Rd Zoo Creek McPherson Zoo Creek Colonial City Dam Clear Fork Kayak West Fork 199 West Fork Confluence 26
27 Mass Balance Study Clear Fork at Trinity Park 27
28 Mass Balance Study Clear Fork at Trinity Park 28
29 Mass Balance Study Clear Fork at Trinity Park 29
30 Trinity River Tracer Study 4 Tracer Compounds Used for Analysis: Acetaminophen (Pain Reliever) Caffeine Sucralose Sulfamethoxazole (Antibiotic) 30
31 Tracer Study Trinity River Tracer Study Concentration ng/l Bryant Irving Rogers Rd Zoo Creek McPherson Zoo Creek Colonial City Dam Clear Fork Kayak West Fork 199 West Fork Confluence Acetaminophen Caffeine Sucralose Sulfamethoxazole 31
32 Total Nitrogen Nitrogen Concentration in the Trinity River mg/l /30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 Bryant Irving Rogers Rd. Zoo Creek McPherson Zoo Creek Colonial City Dam Clear Fork Kayak West Fork 199 West Fork Confluence NH3 NOx Org N 32
33 Total Phosphorus Phosphorus Concentration in the Trinity River mg/l /30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 1/30/2017 Bryant Irving Rogers Rd. Zoo Creek McPherson Zoo Creek Colonial City Dam Clear Fork Kayak West Fork 199 West Fork Confluence Org P OPO4 33
34 Continuing Work. Future Plans: Conduct Similar Study in Summer Months Document Avian Seasonality in Upper Clear Fork Add Nutria DNA to Texas Library Field Investigate Outfalls above Clear Trinity Park Site 34
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