1. Standard Operating Procedure WWSOP03000 Effective Date: 11/01/2015 KENTUCKY WATERSHED WATCH BIOLOGICAL STREAM ASSESSMENT High Gradient

2. This presentation is based on the “Kentucky Watershed Watch High Gradient Biological Stream Assessment Standard Operating Procedure WWSOP03000” SOP statement

3. During high flow, sampling should be postponed until baseline conditions exist Waders and specialized wading boots can be utilized Use extreme caution when wading in streams above the knee Samplers should exhibit caution in and around streams to reduce the threat of a falling injury Secure permission from landowner Be aware of increased risk for frostbite or hypothermia in cold temperatures Do not stick hands or feet under large objects. Exercise caution when handling aquatic life. Some organisms pinch, bite, etc.Do not stick hands or feet under large objects. Exercise caution when handling aquatic life. Some organisms pinch, bite, etc. All water bodies have the potential to be contaminatedAll water bodies have the potential to be contaminated Notify others of your whereabouts and carry a cell phoneNotify others of your whereabouts and carry a cell phone Use caution and be aware of your surroundings and locationUse caution and be aware of your surroundings and location Be Safe!

4. Basic scientific approach to stream water quality evaluation Introduction to Macroinvertebrates and Water Quality Collection and analysis procedures used in this guide are designed to: be affordable be affordable be performed quickly and be performed quickly and easily easily provide hands-on experience provide hands-on experience for individuals, community for individuals, community volunteer groups, and school volunteer groups, and school groups of all ages groups of all ages

5. Streams are constantly changing GRAB SAMPLE Instantaneous water Instantaneous water quality quality Quick process Quick process Specific pollutant Specific pollutant analysis analysis BIOLOGICAL ASSESSMENT Water quality over Water quality over time time Takes more time Takes more time Collective indicator of Collective indicator of pollution pollution VS.

6. Benthic Macroinvertebrates Benthic Occurring at the bottom of a body of water

7. Benthic Macroinvertebrates Macro Macro Visible to the naked eye Invertebrate Invertebrate Lacking a backbone

8. Some mussels and other macroinvertebrates are Threatened or Endangered. Therefore, extra care must be taken to return all organisms to the stream. In addition, any vertebrate organisms (ex. fish, snakes, etc.) must be returned to the stream as soon as you realize they have been captured. Important!

9. Macroinvertebrate Habitat Macroinvertebrates have the same basic requirements as humans to thrive They need: Oxygen to breathe Oxygen to breathe Food to eat Food to eat Shelter from Shelter from predators predators Clean, pollution Clean, pollution free environment free environment

10. Macroinvertebrates are good indicators of the general health of streams for several reasons: Easy to collect Easy to collect Tied to specific habitats Tied to specific habitats Most are unable to avoid pollution Most are unable to avoid pollution Different sensitivities and responses to pollution Different sensitivities and responses to pollution

11. Stream Types in Kentucky Overall steep slope Overall steep slope Typically originate in hilly Typically originate in hilly terrain terrain More rapid flow More rapid flow Many riffles Many riffles Overall low slope Overall low slope Typically originate in flat Typically originate in flat terrain terrain Slower flow Slower flow No actual riffles No actual riffles Low Gradient High Gradient

12. Pools - The deepest areas with the slowest flow in a stream Riffles - The areas in a stream where there is a drop in elevation that causes the rocks on the bottom of the stream to extend above the surface of the water creating a churning action that causes the surface of the water to look rough and bubbly Runs - The segments of streams that flow quickly out of a riffle or into a pool Pools - The deepest areas with the slowest flow in a stream Riffles - The areas in a stream where there is a drop in elevation that causes the rocks on the bottom of the stream to extend above the surface of the water creating a churning action that causes the surface of the water to look rough and bubbly Runs - The segments of streams that flow quickly out of a riffle or into a pool High Gradient Instream Characteristics

13. Pools - The deepest areas with the slowest flow in a stream Runs - The segments of streams between pools that are shallower and flow more quickly Pools - The deepest areas with the slowest flow in a stream Runs - The segments of streams between pools that are shallower and flow more quickly Low Gradient Instream Characteristics

14. Types of Pollution Comes from entire watershed Comes from entire watershed Nonpoint Source Point Source Single, identifiable source Single, identifiable source

15. Stream Pollution and Disturbance Chemicals Bacteria and Nutrients Physical

16. The Importance of Habitat Biological Assessment and Habitat Assessment go hand in hand Pre-sampling Information and Procedures

17. You will receive materials from program coordinators or by mail You will receive materials from program coordinators or by mail Read instructions upon receipt! (Critical info on sample times included) Read instructions upon receipt! (Critical info on sample times included) You will receive for sampling: InstructionsInstructions Field RecordsField Records Field Data SheetsField Data Sheets Calculation SheetCalculation Sheet Pre-sampling Information

18. Reach Selection A 100 meter reach that contains your grab sampling site and that is representative of the characteristics of the stream should be selected. NOTE: Following periods of very high water, populations of organisms may be temporarily disrupted. Several weeks to a month may be required to reestablish the benthic community.

19. Reach Selection The 100 meter measurement should be taken down the middle of the stream channel (following the path of the stream as closely as safety allows) using the construction line reel/string provided.

20. Assessment Event Scheduling Biological Assessments must be done in when macroinvertebrate groups are most easily found and before water levels drop in small streams MayorJune

21. Macroinvertebrate Collection Procedures Professional macroinvertebrate samples

22. Macroinvertebrate Collection Procedures Watershed Watch Biological Stream Assessment – simplified scientific methods

23. Enter downstream and work your way upstream so you won’t disturb the in-stream habitat you will be sampling

24. Basic equipment needed:

25. High Gradient/Rocky Bottom Stream Sampling Techniques, Habitats and Procedures Rocky bottoms (or hard substrate) will be prevalent in the riffles (and runs), which are a common feature throughout most streams.

26. SAMPLING TECHNIQUES: Kick - accomplished by using toe and/or heel of shoe to dislodge upper layer of cobble or gravel and scrape underlying bed, allowing downstream flow to carry organisms into net

27. SAMPLING TECHNIQUES: Swish - accomplished by placing item in water in first collection pan and moving it vigorously through water several times to dislodge organisms

28. SAMPLING TECHNIQUES: Jab - accomplished by thrusting D-frame net into targeted habitat in a jabbing motion for approximately 0.5 m (18 in.) and then sweeping water in jabbed area with net two or three times to collect the dislodged organisms

29. SAMPLING TECHNIQUES: For each sampling technique used: In each habitat type sampled, vigorously disturb habitat until: Habitat surface is thoroughly disturbed Habitat surface is thoroughly disturbed No more materials are dislodged into your net No more materials are dislodged into your net (For riffle) all materials on top of stream bed (For riffle) all materials on top of stream bed have been disturbed in sample location have been disturbed in sample location

30. SAMPLING TECHNIQUES: For each technique in each habitat type sampled, use the same level of effort to perform the sample collection.

31. What to do if you collect large quantities of small or fine material (such as sand or silt)? To speed up the process of inspecting this material for macroinvertebrates: Use a bucket and a rinsed net Use a bucket and a rinsed net Half-fill bucket with stream water Half-fill bucket with stream water Place small handfuls of fine material into bucket Place small handfuls of fine material into bucket Vigorously swirl water in bucket to separate Vigorously swirl water in bucket to separate macroinvertebrates from fine material macroinvertebrates from fine material Immediately after swirling, pour water only (not fine Immediately after swirling, pour water only (not fine material) through net to capture any suspended material) through net to capture any suspended macroinvertebrates and place them in first pan macroinvertebrates and place them in first pan Pour the water in an area away from your collection pans Pour the water in an area away from your collection pans Repeat until all of fine material has been processed Repeat until all of fine material has been processed SAMPLING TECHNIQUES:

32. HABITAT TYPES: The following slides describe the possible Habitat Types to be sampled and identify techniques to be used for each:

33. Riffles

34. Riffles Place net on substrate in middle area of riffle where water is flowing fastest Place net on substrate in middle area of riffle where water is flowing fastest Kick a 0.25m 2 (18 in. x 18 in.) area upstream of net Kick a 0.25m 2 (18 in. x 18 in.) area upstream of net Large rocks, leaves and sticks should be hand washed into net Large rocks, leaves and sticks should be hand washed into net

35. Root Mats Associated with Undercut Banks

36. Place large root wad into D-frame dip net and shake vigorously Place large root wad into D-frame dip net and shake vigorously For undercut banks without root mats, use jab For undercut banks without root mats, use jab

37. Aquatic Vegetation (Submerged and Emergent)

38. Use modified jab, thrusting dip net into vegetation for approximately 1 meter (jab below water surface from bottom to top of Use modified jab, thrusting dip net into vegetation for approximately 1 meter (jab below water surface from bottom to top of vegetation) and sweeping through area to collect dislodged organisms

39. Bedrock or Slab-Rock

40. Place edge of dip net flush on substrate with opening facing upstream Place edge of dip net flush on substrate with opening facing upstream Kick approximately Kick approximately 0.1 m 2 (11 in. x 11 in.) of area immediately upstream of net

41. Leaf Packs

42. Take a large handful from each type of area Take a large handful from each type of area present (i.e., riffles, runs and pools) and place into net NOTE: Collect from “conditioned” (i.e., beginning to break down) material Leaf Packs

43. Large Submerged Wood

44. Jab pieces of submerged wood, ranging from roughly 3 to 6 meters in length (10 to 20 linear feet) and ranging from 5–15 cm (2–6 inches) in diameter Jab pieces of submerged wood, ranging from roughly 3 to 6 meters in length (10 to 20 linear feet) and ranging from 5–15 cm (2–6 inches) in diameter

45. Silt, Sand, and Fine Gravel (Submerged)

46. Sample this habitat last Sample this habitat last Scoop a double handful of substrate by spreading hands 1 foot apart and bringing both hands together in a sweeping motion at a depth of 2 to 3 inches into the substrate Scoop a double handful of substrate by spreading hands 1 foot apart and bringing both hands together in a sweeping motion at a depth of 2 to 3 inches into the substrate Place material into first collection Place material into first collection pan and work through with fingers to inspect (or use Bucket method) NOTE: This substrate consists of sediment with particle sizes that are larger than clay and smaller than a BB Silt, Sand, and Fine Gravel (Submerged)

47. SAMPLING PROCEDURES: The first time you conduct this assessment, select and measure a 100 meter reach containing your grab sampling site and The first time you conduct this assessment, select and measure a 100 meter reach containing your grab sampling site and representative of stream characteristics Return to this same reach for all Return to this same reach for all future assessments Sampling begins at downstream Sampling begins at downstream end of reach and proceeds upstream

48. SAMPLING PROCEDURES: Fill first collection pan at least half full of stream water (if it is too full, organisms may slosh out) Fill first collection pan at least half full of stream water (if it is too full, organisms may slosh out) Place a small amount of water in the second collection pan Place a small amount of water in the second collection pan

49. SAMPLING PROCEDURES: Kick in four riffle locations (divide among various riffles in reach) Kick in four riffle locations (divide among various riffles in reach) Sample once in each of other Habitat Types, if present: Sample once in each of other Habitat Types, if present: Undercut Banks/Root Mats Jab Undercut Banks/Root Mats Jab Aquatic Vegetation Jab Aquatic Vegetation Jab Bedrock or Slab-rock Kick Bedrock or Slab-rock Kick Leaf Packs Swish Leaf Packs Swish Large Submerged Wood Jab Large Submerged Wood Jab Silt, Sand, & Fine Gravel Scoop Silt, Sand, & Fine Gravel Scoop

50. SAMPLING PROCEDURES: After sampling each location, transfer any collected material from net (or hands for sand, silt, and fine gravel) to the first collection pan – After sampling each location, transfer any collected material from net (or hands for sand, silt, and fine gravel) to the first collection pan – invert net to transfer Swish inverted net and Swish inverted net and inspect for organisms. Forceps may be needed to remove organisms from net. Place any organisms found into second collection pan NOTE: Samples from all individual sampling spots are combined to make one composite sample for reach

51. SAMPLING PROCEDURES: Remove large debris from first collection pan, swish and discard after inspecting debris for organisms Remove large debris from first collection pan, swish and discard after inspecting debris for organisms Many of these organisms may Many of these organisms may be quite small – take time to swish and thoroughly inspect the materials before discarding Pick any macroinvertebrates from first collection pan into second collection pan – avoid transferring debris Pick any macroinvertebrates from first collection pan into second collection pan – avoid transferring debris

52. If first collection pan becomes full or muddy: Transfer all macroinvertebrates collected to that point into second collection pan Transfer all macroinvertebrates collected to that point into second collection pan After all macroinvertebrates are After all macroinvertebrates are removed from first collection pan, discard water and replace with new water from stream Continue sampling and placing Continue sampling and placing new material collected in first pan until reaching required number of samples Repeat as needed Repeat as needed SAMPLING PROCEDURES:

53. Inspect first collection pan to ensure all organisms have been transferred to second collection pan Inspect first collection pan to ensure all organisms have been transferred to second collection pan Using “Kentucky Water Watch Benthic Macroinvertebrate Identification Key”, identify and count organisms in second pan Using “Kentucky Water Watch Benthic Macroinvertebrate Identification Key”, identify and count organisms in second pan Count only living organisms (empty shells should Count only living organisms (empty shells should not be counted)

54. SAMPLING PROCEDURES: As you identify organisms As you identify organisms in second collection pan, record your count on “Kentucky Watershed Watch Biological Assessment Field Data Sheet” (Field Data Sheet) Stop counting when you reach 31 for any one type of organism. If more than 50, write “Note: 50+” in the box.

55. Macroinvertebrate Identification

61. Use key to identify on your own

62. Macroinvertebrate Identification

64. Return organisms to stream as soon as Field Data Sheet is completed SAMPLING PROCEDURES:

65. Record each habitat type sampled in reach and any related comments on the “Kentucky Watershed Watch Biological Assessment Field Record” (Field Record) Record each habitat type sampled in reach and any related comments on the “Kentucky Watershed Watch Biological Assessment Field Record” (Field Record) NOTE: Percent of Reach is not required for high gradient streams

66. SAMPLING PROCEDURES: Place thermometer at mid-depth of stream in location with constantly flowing water Place thermometer at mid-depth of stream in location with constantly flowing water Leave thermometer there until reading stabilizes Leave thermometer there until reading stabilizes Record stream temperature in degrees Celsius (ºC) on your Field Record Record stream temperature in degrees Celsius (ºC) on your Field Record

67. POST-SAMPLING PROCEDURES: After biological sampling has been completed, conduct the Kentucky Watershed Watch Habitat Assessment After biological sampling has been completed, conduct the Kentucky Watershed Watch Habitat Assessment Conduct Habitat Assessment with another trained person, if possible Conduct Habitat Assessment with another trained person, if possible Complete all Biological Assessment forms after returning home Complete all Biological Assessment forms after returning home

68. Transfer all Abundance Count numbers from the Field Data Sheet to the column labeled “Abundance Count” on the Calculation Sheet Transfer all Abundance Count numbers from the Field Data Sheet to the column labeled “Abundance Count” on the Calculation Sheet Completing Your Forms

69. Transfer the Biotic Index number on the Calculation Sheet to the box labeled “Biotic Index” on yourTransfer the Biotic Index number on the Calculation Sheet to the box labeled “Biotic Index” on your Field Record Completing Your Forms

70. Find the Bioregion for your site in the “Bioregion” box located on your Field RecordFind the Bioregion for your site in the “Bioregion” box located on your Field Record Completing Your Forms

71. Once your Bioregion is determined, use the Biotic Index number and the “Integrity Rating Chart” from page two of Calculation Sheet to determine the Integrity Rating for your site and record in the box labeled “Integrity Rating”Once your Bioregion is determined, use the Biotic Index number and the “Integrity Rating Chart” from page two of Calculation Sheet to determine the Integrity Rating for your site and record in the box labeled “Integrity Rating” Completing Your Forms

72. Following each Biological Assessment event, please mail a copy of all completed forms to: KY Water Watch Office 200 Fair Oaks Lane, 4 th Floor Frankfort, KY 40601 Here the forms will be recorded in the KY Water Watch database and filed for safekeeping If you have any questions or difficulties, please contact the KY Water Watch office at: 1-800-928-0045 FORM SUBMITTAL

73. The Watershed Watch Biological Stream Assessment analyzes the abundance and pollution tolerance of all macroinvertebrate groups collected to make conclusions about the health of the aquatic system. INTERPRETATION

74. Different macroinvertebrate groups have different sensitivities and responses to pollution. These are referred to as pollution tolerance. POLLUTION TOLERANCE Intolerant Tolerant

75. TOLERANCE VALUE (Intolerant) (Tolerant) Tolerance Value Value Stonefly = 2 Leech = 9

76. The number of organisms from a macroinvertebrate group present in the stream is referred to as abundance. ABUNDANCE Abundance of Aquatic Worms and other tolerant organisms indicate organic pollution

77. ABUNDANCE VALUE Abundance Count CountRange= >30 (abundant) 30-5 (common) 4-1 (rare) 1832Abundance Value = Value = (abundant) 6 (common) 3 (rare) 1 Example:

78. The different areas resulting from the influences of Kentucky’s geology are called bioregions. The four bioregions are the: Mountains (MT) Mountains (MT) Bluegrass (BG) Bluegrass (BG) Pennyroyal (PR) Pennyroyal (PR) Mississippi Valley/Interior River Lowland (MVIR) Mississippi Valley/Interior River Lowland (MVIR) BIOREGIONS

79. BIOTIC INDEX Total Tolerance Total Abundance = Biotic Index The Biotic Index is a number that represents all of the information collected in the biological assessment of your stream reach

80. INTERPRETING DATA STATEWIDE

81. Phase 2 (Biological and Habitat Assessment) Phase 2 (Biological and Habitat Assessment) Training: Training: Your sampling and assessment skills are considered Your sampling and assessment skills are considered current after completing WWKY-approved training current after completing WWKY-approved training You will be notified that training updates are You will be notified that training updates are required if modifications are made to WWKY required if modifications are made to WWKY methods methods RECERTIFICATION

82. IMPORTANT: If you observe an outfall, pipe, or anything that might indicate illegal pollution of a waterway, contact KDOW to report complaints or concerns at 502-564-3410 during business hours. For environmental emergencies such as spills of gas, oil or other substances, contact the Environmental Response Team at 502-564-2380 or 1-800-928-2380.