1. Measuring Stream Microbiology:
Preliminary Results
Dr. Robert D. Simon
Mr. Jacob D. Almeida
Mr. Jonah Stevens
Department of Biology
SUNY-Geneseo

2. Microbial Quality is Monitored by Measuring Surrogates
Coliforms
Fecal Coliforms
Escherichia coli Fecal Streptococci
Enterococci
Total Heterotrophic Bacteria

3. Coliforms Found in the gut and feces of warm-blooded animals
Aerobic and facultative anaerobic, Gram-negative, non-spore forming rod shaped bacteria that ferment lactose with gas formation within 48 hours at 35 °C.
Enterobacteriaceae
Membrane Filtration (MF) MF - Dark colony with a metalic sheen on an “Endo” type medium.

4. Fecal Coliforms A subset of Total Coliforms
Fecal coliforms - Produce gas from lactose at 44.5 °C.
Prior to its 1986, EPA recommended the use of fecal coliforms as an indicator organism to protect bathers from gastrointestinal illness in recreational waters.

5. Membrane Filter methods for Total Coliforms and E
Membrane Filter methods for Total Coliforms and E. coli using Millipore mColiBlue24
Next day results
Easy to read with turbid samples
Clonal isolates can be saved for further study

6. Fecal Streptococcus Isolated from the feces of warm-blooded animals
A number of species of the genus Streptococcus
It is not possible to differentiate the source of fecal contamination based on speciation
FC/FS ratio can also not be used as a means of differentiating human and animal feces

7. Enterococcus
A subgroup of fecal streptococci, including: S. faecalis, S. faecium, S. gallinarum, S. avium
Differentiated by ability to grow in 6.5% NaCl at 10 °C and 45 °C
Difco m Enterococcus Agar

8. Heterotrophic Bacteria
Bacto R2A Agar. A low-nutrient medium. Incubation at 25 °C for hours, stimulates the growth of stressed and chlorine-tolerant bacteria
Multiple 20 l drops of water dilutions are spotted on plates, and micro-colonies are counted under a dissecting microscope

9. EPA’s Recommended 1986 Water Quality Criteria for Bacteria in Recreational Fresh Water
Based on the epidemiology of water-born disease occurrence as E. coli and Enterococci levels exhibit a strong correlation to swimming-associated gastroenteritis.

10. Results

11. Two Types of Stream Flow
Regular Flow (Grab Samples)
Slow flow during the summer, a portion probably from groundwater
It is surprising how much flow there is in the Winter
Weekly measure stream characteristics
Events
Due to rain or snow melt

12. Assessing Overall Stream Microbial Quality From Grab Samples
Because of the variability in individual measurements, it is valuable to to combine monthly regular (non-event) estimates.
The geometric means of monthly samples are calculated because this damps out the effect of a few extreme values.

13. Graywood Sand Point Long Point Long Pt. McPhersons Point Cottonwood
Experimental sites
with weed beds
Control sites
with weed beds
Graywood
Experimental site
without weed beds
Control sites
without weed beds
Sand Point
Long Point
Long Pt.
Cove
McPhersons Point
Cottonwood
Gully
Sutton Point
Southwest
Creeks
North
McMillan

14. Conclusions about Regular Flow
E. coli and Enterococcus levels are seasonal
Total Coliforms and Heterotrophic Bacteria vary little over the year.
Graywood is the most regularly contaminated stream and on the western side of the lake the streams at the northern end have the most problems with water quality.

15. Events
The Number and Characteristics, of Events vary by Season and Stream Source

16. Manure Spread on Snow Above Graywood - February 2003

17. Events
There are Different Types of Events

18. Sutton Point – Two Recorded Events from 6/17/03 – 9/2/03

19. Given the Large Inputs into Streams During Events Do these Persist in the Stream or Lake?
Sediment Microbiology
Analysis of Fecal Coliform Levels in Sediments from the Stream and Lake Near the Stream Outfalls
No Evidence for Significant Accumulations of Fecal Material in Sediments or Stream Bottoms

20. Major Conclusion

21. Future Plans Continue monitoring program
Use current data to better understand the number, types and magnitude of different events. This may aid in monitoring clean-up as one might aim to prevent certain event types.
Examine the changing character of the Heterotrophic bacterial flora

22. The End