Standard Operating Procedure WWSOP02000 Effective Date: 04/01/2013

1. KENTUCKY WATERSHED WATCH STREAM GRAB SAMPLE COLLECTION Standard Operating Procedure WWSOP02000 Effective Date: 04/01/2013

2. Grab Sample SOP This presentation is based on the “Kentucky Watershed Watch Stream Grab Sample Collection Standard Operating Procedure WWSOP02000” and is available from wwky.org

3. Stream Grab Sample Collection Procedures • Stream safety • Sample site • location • Pre-sampling • information • Collection procedures • Preservation • Recordkeeping • Transportation • Results

4. Sample Site Location • Site Coordinates and Photos • GPS (decimal degrees – 5 decimal places) • OR Digital map (KGS WW Site - http://kgs.uky.edu/kgsmap/krww/viewer.asp • or Google Earth) • Take photos upstream and downstream of sampling point – with recognizable landmarks; should be taken annually during Summer sampling event • Label photos: “Site#upstream” and “Site#downstream” • Submit information to Water Watch office Email: OllieTheOtter@ky.gov • Phone: 1-800-928-0045

5. Pre-sampling Information • You will receive for sampling: • Instructions • Transport information • Chain of Custody Records • Gloves (disposable) • Sample containers • Preservative (if required) • You will receive materials from program coordinators or by mail • Read instructions upon receipt! (Critical info on sample times and delivery included) • Inspect all sample and preservative containers upon receipt for damage or breakage

6. Pre-sampling Information Sample containers vary for the job at hand

7. Pre-sampling Information Watershed Watch Chain of Custody Record The sampling containers are pre-marked with a unique Site Number The Site Number should match the number on the “Chain of Custody Record”

8. Pre-sampling information Ireceived a bottle with a site number I don’t recognize! Check the documents in your packet, you may have been randomly assigned to collect a blank or a duplicate

9. Pre-sampling Information • Blank - A sample bottle filled while at the stream site with distilled water • Serves as a check for possible contamination during sample handling or processing at the lab • DO NOT indicate that the sample is a blank – the lab should not know

10. Pre-sampling Information • Duplicate - A sample bottle filled with stream water at your site at the same time as your regular sample • Serves as a check for variability among samples that should yield approximately the same lab results • DO NOT indicate that the sample is a duplicate – the lab should not know

11. Pre-sampling Information • Proper attire for handling samples • Gloves (disposable) • Eye protection

12. Collection Procedures Basic equipment needed

13. Collection Procedures Selecting your sample collection location On your first visit to your site, select an area of the stream where the flow is concentrated and where you can completely submerge and fill the bottle without touching the bottom NOTE: It is important that you collect your sample at the same location each time

14. Collection Procedures Concentrated Flow Location

15. Collection Procedures Sample Collection Location High Gradient

16. Collection Procedures Sample Collection Location Low Gradient

17. Collection Procedures Grab Sample Collection • Enter stream at a downstream location • Walk upstream to avoid disturbing bottom • Collect sample at the same location each time in • the middle of your concentrated flow location • and at mid-depth of stream

18. Collection Procedures Grab Sample Collection • 0 = Dry • 1 = Ponded • 2 = Low • 3 = Normal • 4 = Bank Full • 5 = FLOOD! Flow Rate • Do not sample • during these • flow conditions

19. Collection Procedures ForE. coli grab sample collection: • Put on disposable gloves • DO NOT RINSE BOTTLE • Point mouth of bottle directly over surface of water, dip • and scoop bottle upstream beneath water surface, and • through water column in one swift motion to fill above • line marked on bottle • If preservative is in bottle, take caution to angle mouth so • as not to spill any while collecting sample • Cap container, leaving at least a small airspace at top to • allow lab mixing • Place in wet ice immediately

20. Collection Procedures For grab sample collectionother thanE. coli: • Put on disposable gloves • Rinse bottle with stream water three times • (fill bottle ¼ full of stream water, cap, shake, and discard • downstream) • Point mouth of bottle upstream (against the flow), • submerge entire bottle to mid-depth of stream, • scoop and fill • Rinse cap in stream and cap bottle • Add preservative (if necessary) and place in wet ice

21. Equipment Chemical Safety • What to do if you get a chemical on you • For emergency first aid in the field, flush • affected area with large amounts of water • and seek medical attention immediately • Follow the first aid procedures found on • the Material Safety Data Sheets (MSDS)

22. Preservation • Samples must be kept immersed in wet ice • (but not frozen) until received by laboratory • Some samples may require other preservatives • (check your sample event packet)

23. Preservation Like containers, preservation methods vary (You don’t have to memorize these)

24. Preservation Sample Holding Time: • length of time a preserved sample can be stored • after collection and before it is analyzed without • significantly affecting the results • from the minute you collect a sample, the • clock starts ticking on the holding time • varies for different types of analysis • established by the US Environmental • Protection Agency

25. Preservation Sample Holding Times E. coli= 6 hours maximum Other thanE. coli= Follow holding time instructions received in your sampling event packet

26. Recordkeeping Watershed Watch Chain of Custody Record The Chain of Custody Record (COC) serves to: • document and record the • transfer of the samples • from the stream to the • laboratory • function as a field • measurement form • provide a place for field • observations

27. Recordkeeping • Site # • Location description • Sample collector name • and contact info • Correct any preprinted • information if necessary Chain of Custody Record prepopulated information Watershed Watch Chain of Custody Record

28. Recordkeeping Volunteer entries on the Chain of Custody Watershed Watch Chain of Custody Record • Date and Time of sample collection • Field Observations and Measurements • Date and time of sample • Relinquished/Received • must be identical Cross through any mistakes and initial – do not erase

29. Transportation • 1. Make delivery arrangements • before the sampling event • 2. Take your sample and COC • to the designated drop-off • location • 3. Double-check your • recordkeeping paperwork • 4. Make sure to sign your COC Sample Delivery

30. Transportation Runners Collection 7:00-10:00 AM 9:00-1:00 PM 6 hrs (E. coli) Sampler Sampler Sampler Drop Sampler Runner Off LAB Sampler

31. Results Professional labs analyze samples

32. Results Bacteria Analysis (Older Method) • Colony Forming Units (CFU) • Lab processes your stream sample and applies it to a special plate • Plate is kept warm for a specified period of time • Colonies of microscopic bacteria that have formed are counted • Results are recorded in CFU

33. Results E. coli Analysis (Newer Method) Add Colilert to sample and dissolve Count fluorescent wells and convert Pour sample into Quanti-Tray Seal tray and keep warm for 24 hours

34. RESULTS Results will be posted on the Kentucky Geological Survey website after the basin data manager and KY Watershed Watch have reviewed for errors http://kgs.uky.edu/kgsmap/krww/viewer.asp

35. Interpretation E. coli • shortened form of Escherichia coli • microscopic bacteria that live in the • intestines of warm-blooded animals • and are passed in fecal material • used as an indicator of the possible • presence of pathogens that may cause • infection or illness Enlarged photo of E. coli bacteria

36. Interpretation Sources of E. coli in Streams

37. Interpretation Surface Water Quality Standards for E. coli • Swimming Standard of 240 cfu/100 ml (single grab sample) • Standards apply during Recreation Season from May 1 to October 31

38. Interpretation Nutrients • generally refers to forms of • nitrogen (N) and phosphorus (P) • excess nutrients in streams can • cause overgrowth of algae and • plants, which disrupts healthy • natural processes • this can lead to large decreases • in dissolved oxygen, a condition • called eutrophication

39. Interpretation Nutrients • Excess of nutrients (N & P) • Abundant growth of algae • Algae die • Decomposition of algae by bacteria consumes oxygen • Low dissolved oxygen levels can lead to fish kills EUTROPHICATION

40. Interpretation Sources of Nutrients in Streams

41. Interpretation Surface Water Quality Standards for Nutrients • Kentucky does not have numeric surface water standards in regulation for all nutrients of concern (including • Total Nitrogen and Total Phosphorus) • Surface water standards in Kentucky regulations include: • Ammonia (un-ionized) 0.05 mg/l (parts per million) • Nitrate (NO3) 10 mg/l • Sulfate (SO4) 250 mg/l

42. Interpretation Metals • elemental substances that are good • conductors of heat and electricity • naturally occurring in rock and soil, and • can be weathered and carried to • streams when exposed • can be harmful to humans or aquatic life • at low levels over a long time (chronic) • or at higher levels for a short time • (acute) Heavy Metal

43. Interpretation Sources of Metals in Streams

44. Interpretation Surface Water Quality Standards for Metals • Aluminum • Antimony • Arsenic • Barium • Beryllium • Boron • Cadmium • Calcium • Chromium • Cobalt • Copper • Gold • Iron • Lead • Lithium • Magnesium • Manganese • Nickel • Phosphorus • Potassium • Selenium • Silicon • Silver • Sodium • Strontium • Sulfur • Thallium • Tin • Vanadium • Zinc Underlined metals are ones for which Kentucky has numeric water quality standards, which can be found in 401 KAR 10:031 Section 6.

45. Interpretation Metals • A fish consumption advisory is in effect statewide • All Kentucky waters are under advisory for mercury. Women of childbearing age and children 6 years of age or younger should eat no more than one meal per week of freshwater fish. Adult men and other women are not included in the consumption notice. • This is not an emergency as organic mercury can occur • naturally in the environment and does not affect swimmers, skiers or boaters. Fish can accumulate these low levels of mercury by eating plankton and other small aquatic creatures.

46. Interpretation Pesticides • chemicals used to kill undesirable plants • or insects • improper timing of application and • overuse can lead to runoff into streams • can be harmful to humans or aquatic life • at low levels over a long time (chronic) • or at higher levels for a short time • (acute)

47. Interpretation Sources of Pesticides in Streams

48. Interpretation Surface Water Quality Standards for Pesticides • Kentucky does not have numeric water quality standards for all pesticides (including atrazine) • Surface water standards in Kentucky regulations include: • 2,4-D 0.1 mg/l (parts per million)

49. (CFU)