Dr. Robin Brinkmeyer. Department of Marine Sciences. Dr. John Schwarz Department of Marine Biology. Texas A&M University at Galveston
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1 Population Dynamics of Escherichia coli and Enterococcus spp. in Buffalo Bayou and White Oak Bayou Dr. Robin Brinkmeyer Dr. Rainer Amon Department of Marine Sciences Dr. John Schwarz Department of Marine Biology Texas A&M University at Galveston
2 A comprehensive, multi-year study of the fate and transport of E. coli and Enterococcus spp. to address several questions: 1. Under what conditions can E. coli and Enterococcus spp. bacteria survive for extended periods in the natural waters and sediment in Buffalo Bayou and White Oak Bayou and in the soils in the watersheds? 2 Under what conditions can E coli and Enterococcus spp 2. Under what conditions can E. coli and Enterococcus spp. bacteria replicate (grow) in natural waters and soils in Buffalo Bayou and White Oak Bayou?
3 Questions 1& 2 -Sampled from January to August 28 to assess influence of temperature and water flows Analysis of nutrients in water, sediments (from pore water) and soils FISH and Micro-FISH (microautoradiography combined with FISHdetermines activity) analysis of all substrates with E. coli and Enterococcus specific DNA probes (in progress) Traditional detection methods for comparison Laboratory incubations using substrates to determine if E. coli and Enterococcus will replicate or are only viable but non culturable (VBNC) (in progress and focus of FY29) Determination of false-positives produced with traditional FC detection methods using DNA probes (in progress) HFERP fingerprinting to determine naturalized indicator bacteria (in progress)
4 3. Under the conditions indicator bacteria are found to replicate in natural waters and sediment, what are they using for growth substrates? Are they utilizing WWTP effluent derived substrates from chlorine or UV based processes or from other substrates found in the waters and sediments? End member chemical characterization of organic sources (awaiting results from contract lab) Stable isotopic composition of bacteria that replicate in natural substrates (in progress)
5 Typical δ15 N values of various nitrogen sources in the environment.
6 4. Are these indicator bacteria attached to particulate matter in the watersheds or surviving on biofilms in discharge conveyance systems and are then flushed into bayous during rain events? FISH analysis of biofilms and particulate t matter collected during storm events (focus of FY29) Determination of biofilm growth on microscope slides affixed in discharge conveyance systems (in progress)
7 5. How does the hydrologic cycle influence survival and replication of these indicator bacteria? Sampling during storm events. Analyses to include nutrients, DOC, FISH, IDEXX Comparison of strain genotypes from soils and sediments to cultivated isolates. Focus of FY29
8 WWTP, Water, Sediment & Soil Samples (base flow & rain events) IDEXX Colilert EPA Membrane Filter methods Isolate Cultures DNA fingerprinting Incubations with filtered water using E. coli & Enterococcus isolated from same sample Carbon, Nitrogen, Sulfur stable isotope analyses of bacteria and substrates Stormwater biofilm cultivation experiments Detection & enumeration of E. coli and Enterococcus with fluorescence in situ hybridization (FISH)
9 2 Control Sites on Lake Creek in the Caney Creek Watershed
10 Year 1 Sampling Sites
11 In Spring 28, learned that FY29 funding would be reduced by more than 5%. Increased FY28 budget to include 15 additional sampling sites and QPCR & DNA probing analyses. Trade offsmore samples but less time in FY28 for processing and FY29 reduction in research staff.
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14 Sediment Cores collected Above the Water Line (AWL), at Water Line (WL) & Below Water Line (BWL). Three horizons sub-sampled: top (-1cm), mid (variable), bottom (variable). 5/21/ Sediment Cores (AWL) 5/21/ Sediment Cores (WL) 5/21/ Sediment Cores (BWL) MPN/g Wet Weig (24-25cm) (14-15cm) 15cm) (-1cm) MPN/g Wet Weig (16-17cm) (8-9cm) (-1cm) g MPN/g Wet Wei (24-25cm) (14-15cm) (-1cm) 8/6/ Sediment Cores (AWL) High Flow 8/6/ Sediment Cores (WL) High Flow 8/6/ Sediment Cores (BWL) High Flow MPN/g Wet Weig (12-13cm) (-1cm) (19-2cm) MPN/g Wet Weig (11-12cm) (-1cm) (24-25cm) MPN/g Wet Weig (1-11cm) (-1cm) (2-21cm) Highest bacterial concentrations typically in top (-1 cm) horizon
15 Buffalo Bayou (sediment cores- top horizons) Sediment Core (WL) Sediment Core (AWL) Sediment Core (AWL) MPN/g Wet WT (3. cfs) MPN/g Wet WT MPN/g Wet WT Water MPN/1ml Water MPN/1ml Water MPN/1ml MPN/1 ml (3. cfs) MPN/1 ml MPN/1 m
16 Buffalo Bayou (sediment cores- top horizons) Sediment Core (WL) Sediment Core (WL) 1136 Sediment Core (AWL & WL) WT MPN/g Wet (92 cfs) MPN/g Wet WT Ec 4212 (227 cfs) WT MPN/g Wet (22 cfs) Water MPN/1ml Water MPN/1ml 1136 Water MPN/1ml MPN/1ml LowFlow LowFlow (212 cfs) (227 cfs) MPN/1ml MPN/1ml (22 cfs)
17 Buffalo Bayou (sediment cores- top horizons) Sediment Core (AWL) Sediment Core (BWL) Sediment Core (BWL) MPN/g Wet WT High Flow MPN/g Wet W MPN/g Wet W High Flow Water MPN/1ml Water MPN/1ml Water MPN/1ml PN/1ml M High Flow MPN/1ml MPN/1ml High Flow BB at 61 BB at Shepherd
18 Buffalo Bayou (sediment cores- top horizons) Sediment Core (WL) Sediment Core (BWL) Sediment Core (WL) MPN/g Wet WT MPN/g Wet W MPN/g Wet WT 1 5 High Flow (193.8 cfs) Water MPN/1ml Water MPN/1ml Water MPN/1ml MPN/1ml l MPN/1ml l MPN/1m High Flow (193.8 cfs) BB at Main St.
19 White Oak Bayou (sediment cores- top horizons) Sediment Core (WL) Sediment Core (BWL) Sediment Core (BWL) MPN/g Wet WT ( cfs) Water MPN/1ml MPN/g Wet WT Water MPN/1ml MPN/g Wet WT Water MPN/1ml ( cfs) MPN/1ml ( cfs) MPN/1ml MPN/1ml ( cfs)
20 White Oak Bayou (sediment cores- top horizons) Sediment Core (BWL) MPN/g Wet WT (27.13 cfs) Water MPN/1ml Water MPN/1ml 1139 Water MPN/1ml MPN N/1ml (27.13 cfs) MPN N/1ml (11.85 cfs) MPN N/1ml (15.33 cfs)
21 White Oak Bayou (sediment cores- top horizons) Water MPN/1ml Water MPN/1ml Water MPN/1ml MPN/1ml High Flow (43.5 cfs) MPN/1ml (35.7 cfs) MPN/1ml Little White Oak Bayou (sediment cores- top horizons) Sediment Core (AWL) Sediment Core (WL) MPN/g WetW ( cfs) Water MPN/1ml MPN/g Wet WT Water MPN/1ml High Flow (3. cfs) 1ml MPN/ ( cfs) MPN/1 1ml High Flow (3. cfs)
22 Buffalo Bayou below confluence with White Oak Bayou Sediment Core (WL) MPN/g Wet W Water MPN/1ml MPN/1 ml
23 Control Sites- MPN values in sediment and water Control Site 1 Sediment Core (AWL) Control Site 2 Sediment Core (BWL) MP PN/g Wet WT MPN/g Wet WT ( cfs) Control Site 1 Water MPN/1ml Control Site 2 Water MPN/1ml ml MPN/ MPN/1 1ml ( cfs)
24 Station Buffalo Bayou at 61- Cross section analysis of sediments 6/4/ : Sediment Core Stream Cross Section Core 1-top Core Core 1-mid 1-bot Core 2-top Core 2 -mid Core 2-bot Core 3- top Core 3-mid Core 3-bot Core 4-top Core 4-mid Core 4-bot Core 5-top Core 5-mid Core 5-bot Core 6-top Core 6-mid Core 6-bot Core 7-top Core 7-mid Core 7-bot Core 8-top Core 8-mid Core 8-bot Core 9-top Core 9-mid Core 9-bot Core 1-top Core 1-mid Core 1-bot Core 11-top Core 11-mid Core 11-bot Dry rim of bayou Midway in bayou
25 Station Buffalo Bayou at Shepherd- Cross section analysis of sediments 6/1/ : Sediment Core Stream Cross Section MPN/g Wet Weigh ht Trib Core 1-top Core 1-bot Core 2-mid Core 3-top Core 3-bot Core 4-mid Core 5-top Core 5-bot Core 6-mid Core 7-top Core 7-bot Core 8-mid Core 9-top Core 9-bot Core-mid Grab 1 Grab 3 Dry rim of bayou Midway in bayou
26 Rake Study Disturbed sediments at particle size fractionation & transport analysis 6/18/ : E. coli MPN/1ml Rake Study (m) 6/18/ : E. coli MPN/1ml Rake Study (5m) 6/18/ : E. coli MPN/1ml Rake Study (1m) MPN/1ml (Before (After MPN/1ml (Before (After MPN/1ml (Before (After >63um 63-25um <25um Particle Size Fractionation 25-1um <1um >63um 63-25um <25um Particle Size Fractionation 25-1um <1um >63um 63-25um <25um Particle Size Fractionation 25-1um <1um 6/18/ : Enterococcus MPN/1ml Rake Study (m) 6/18/ : Enterococcus MPN/1ml Rake Study (5m) 6/18/ : Enterococcus MPN/1ml Rake Study (1m) MPN/1ml (Before (After MPN/1ml (Before (After MPN/1ml (Before (After >63um 63-25um <25um Particle Size Fractionation 25-1um <1um >63um 63-25um <25um Particle Size Fractionation 25-1um <1um >63um 63-25um <25um Particle Size Fractionation 25-1um <1um 6/18/ : Total Coliforms MPN/1ml Rake Study (m) 6/18/ : Total Coliforms MPN/1ml Rake Study (5m) 6/18/ : Total Coliforms MPN/1ml Rake Study (1m) MPN/1ml (Before (After MPN/1ml (Before (After MPN/1ml (Before (After >63um 63-25um <25um Particle Size Fractionation 25-1um <1um >63um 63-25um <25um Particle Size Fractionation 25-1um <1um >63um 63-25um <25um Particle Size Fractionation 25-1um <1um
27 Sediment analysis after Hurricane Eduard Eduard - 8/6/ : Soil According to Soil Horizon BB at 61 Wet Weight AW WL-TOP AW WL-MID -BOT -TOP AWL- WL- WL- -MID MPN/g WL-B BOT BWL-TO OP BWL-M ID BWL-BOT LC-BOT Eduard - 8/6/ : Soil According to Soil Horizon 9 8 BB at Main St Weight MPN/g Wet AW L- TOP AW L- MID AW L- BOT WL- TOP WL- MID WL- BOT BW L- TOP BW L- MID BW L- BOT LC- BOT
28 Total Suspended Solids (63-25 μm) and Dissolved Organic Carbon TSS higher in Buffalo Bayou- carbon and stable isotope analysis of particle size Fractionation in progress. TSS and DOC of Buffalo/Whiteoak Bayous during High Flow /L) TSS (mg/ TSS DOC 4 3 DOC (um M) WOB WOB WOB BB BB Confluence Sampling Sites
29 Percent sediment moisture vs 3/13/ : % Moisture vs. MPN/g Wet Weight AWL-Top AWL-Mid AWL-Bot WL-Top WL-Mid WL-Bot BWL-Top BWL-Mid BWL-Bot Long-Bot Soil Horizon 1. E. coli MPN/g wet wt 9. Et Enterococci 8. MPN/g wet wt 7. % M o isture Moisture %
30 Analysis of Biofilms Buffalo Bayou: Biofilm Biofilm tree Biofilm brick branch Biofilm plastic
31 White Oak Bayou: Biofilm MPN/g We et Weight Biofilm w ood Biofilm plastic Biofilm glass
32 E. Coli Incubation Experiments in bayou water Very ypreliminary need to be repeated and confirmed with FISH-DNA probing anlaysis Station Water (1E2) Station Water (1E3) 7.E+6 2.5E+6 6.E+6 5.E+6 4.E+6 2.E+6 1.5E+6 3.E+6 1.E+6 2.E+6 1.E E+5.E Hours.E Hours Sediment incubation studies in progress using gamma irradiated samples from 6 sites.
33 FY29 Focus on storm drains, biofilms, and incubation studies
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