31st Annual Meeting New England Association of Environmental

31st Annual Meeting New England Association of Environmental Biologists 31st Annual Meeting Grand Summit Resort Hotel Mount Snow, Vermont March 14, 2007

Using BioSim2 to identify statisticallyvalid sectors of indicator taxa By Darlene Olsen Mathematics Department Norwich University Northfield, VT & Carlos Pinkham Biology Department Norwich University Northfield, VT

Outline Introduction – Index of Biotic Similarity – Brief Review – What is a Sector? VTDEC Ambient Biomonitoring Program – Small, Moderate Gradient (Wadable) Streams – Macroinvertebrate sampling Results – Macroinvertebrate % Composition Data – Environmentally Reasonable Sectors – Statistically Valid Sectors Conclusions

Index of Biotic Similarity (Pinkham-Pearson Index) Barbour et al. (1992) in a systematic comparison of the metrics proposed in EPA's rapid bioassessment protocol (Pfalkin et al., 1989), concluded that B "may be the most appropriate metric to serve as a measure of community similarity."

Index of Biotic Similarity (Pinkham-Pearson Index)

Matrix of B’s Between 11 Habitat Parameters

What is a Sector? original data matrix, rows coded sites/taxa A B C D E F G CPR1 3.8 0.2 0.9 15.7 0.7 6.9 0 CPR2 3.8 0.7 0.7 12.5 6.4 8.7 0 CPR3 8.4 0.5 2 22 3.8 9.6 1.5 CCR1 5.7 3.2 0 0 1.8 0 0 CCR2 7.7 2.1 0 1.1 1.8 0 0 CCR3 9.5 4.7 0 0.5 5 0 0.2

What is a Sector? Row Dendrogram

What is a Sector? original data matrix sites/taxa A B C D E F G CPR1 3.8 0.2 0.9 15.7 0.7 6.9 0 CPR2 3.8 0.7 0.7 12.5 6.4 8.7 0 CPR3 8.4 0.5 2 22 3.8 9.6 1.5 CCR1 5.7 3.2 0 0 1.8 0 0 CCR2 7.7 2.1 0 1.1 1.8 0 0 CCR3 9.5 4.7 0 0.5 5 0 0.2 Original data matrix rearranged in order of the row dendrogram sites/taxa A B C D E F G CCR2 7.7 2.1 0 1.1 1.8 0 0 CCR1 5.7 3.2 0 0 1.8 0 0 CCR3 9.5 4.7 0 0.5 5 0 0.2 CPR2 3.8 0.7 0.7 12.5 6.4 8.7 0 CPR1 3.8 0.2 0.9 15.7 0.7 6.9 0 CPR3 8.4 0.5 2 22 3.8 9.6 1.5

What is a Sector? Original data matrix rearranged in order of the row dendrogram, taxa coded sites/taxa A B C D E F G CCR2 7.7 2.1 0 1.1 1.8 0 0 CCR1 5.7 3.2 0 0 1.8 0 0 CCR3 9.5 4.7 0 0.5 5 0 0.2 CPR2 3.8 0.7 0.7 12.5 6.4 8.7 0 CPR1 3.8 0.2 0.9 15.7 0.7 6.9 0 CPR3 8.4 0.5 2 22 3.8 9.6 1.5

What is a Sector? Taxa Dendrogram

What is a Sector? Original data matrix rearranged in order of the row dendrogram, taxa coded sites/taxa A B C D E F G CCR2 7.7 2.1 0 1.1 1.8 0 0 CCR1 5.7 3.2 0 0 1.8 0 0 CCR3 9.5 4.7 0 0.5 5 0 0.2 CPR2 3.8 0.7 0.7 12.5 6.4 8.7 0 CPR1 3.8 0.2 0.9 15.7 0.7 6.9 0 CPR3 8.4 0.5 2 22 3.8 9.6 1.5 Rearranging original data matrix in double-dendrogram order F C G D E B A CCR2 0 0 0 1.1 1.8 2.1 7.7 CCR1 0 0 0 0 1.8 3.2 5.7 CCR3 0 0 0.2 0.5 5 4.7 9.5 CPR2 8.7 0.7 0 12.5 6.4 0.7 3.8 CPR1 6.9 0.9 0 15.7 0.7 0.2 3.8 CPR3 9.6 2 1.5 22 3.8 0.5 8.4

What is a Sector? Taxa Dendrogram Identifying environmentally reasonable sectors

What is a Sector? Establishing environmentally reasonable sectors

VTDEC Ambient Biomonitoring Program The Vermont Department of Environmental Conservation (VTDEC) is charged with assessing the biological integrity of wadable stream sites throughout Vermont. Specific data are being collected to better refine the biological expectation of very small ( 1-15 km2), low elevation (118-550 ft), eurythermal, wadable streams of moderate gradient in Vermont. These data will be used to evaluate the response of these streams to current agricultural and storm-water management practices, within their watersheds.

Small, Moderate Gradient (Wadable) Streams From this Program 27 streams were selected, 26 in the Champlain Valley, One in the Connecticut Valley One stream was sampled in 2 successive years to provide a reference for the technique. One stream was sampled at two points Thus data from 29 streams were used in this study.

Deer Creek Engelsby Brook Moorehouse Brook Reference

Macroinvertebrate Sampling Collected in late summer-early Fall index period of Sept to Mid Oct Collected from representative locations in a riffle in the stream. Substrate in an area about 1 square meter upstream of a 500 micron mesh, D net is thoroughly disturbed by hand. Four composite samples collected each sample lasting about 30 seconds Two replicates done per stream; preserved in 75% alcohol.

Macroinvertebrate Processing 1) 2) 3) 4) 5) 6) Sample washed and spread evenly over a white, gridded tray with 24 squares Starting with random grid, it and the next 5 consecutive squares are picked clean of macroinvertebrates using a 3 diopter magnifying light. Process is continued if necessary until 300 organisms are picked. Total Number of squares picked is recorded. Picked macroinvertebrates are preserved in 75% alcohol Macroinvertebrates are identified to species or genus, except Oligochaetes (Family)

Macroinvertebrate Results 170 taxa collected at 29 “Sites” Compressed to 91 taxa at 29 Sites by eliminating – taxa which appeared in only one site with a % composition 1.25% (38) – taxa which appeared in only two sites with a maximum % comp 1.2% (23) – taxa which appeared in only three sites with a maximum % comp 1.1% (11) – taxa which appeared in only four sites with a maximum % comp 1% (7)

RESULTS MACROINVERTEBRATE DATA Why % Composition Stream Dendrogram Taxa Dendrogram Reordered Data Matrix – 0’s Included Reordered Data Matrix – 0’s Removed Reordered Data Matrix - Coded Synthesis

Zero’s Present

Zero’s Present Zero’s Removed

Coding The Reordered Data Matrix

Synthesis

Statistical Analysis Creek 1 Creek 2 Creek n taxon k taxon 2 Group 1 taxon 1 Assumptions – The measurements in each site are independent – The relative abundance of taxa follow a normal distribution Independent

Statistical Analysis taxon k taxon 2 Group 1 taxon 1 Calculations Creek 1 Creek 2 x1,k s1,1 s1,2 s1,k taxon k x1,2 taxon 2 Group 2 x1,1 taxon 1 Creek n Creek 1 Creek 2 Creek m x2,1 x2,2 x2,k s2,1 s2,2 s2,k di x1,i x2,i s1,2i s2,2 i n m

Statistical Analysis Calculations Ho: There is a not a significant difference between the percent compositions of taxa in the sites making up Sector 1 & Sector 2. Ha: There is a significant difference between the percent compositions of taxa in the sites making up Sector 1 & Sector 2. Given Ho is true then di N (0,1) so k 2 2 d ( k ). i i 1 The p-value is calculated using the chi-square distribution.

0 0 0 0 0 0 0 4.1 0 0 0 0 0 0 0.2 2.6 0 0 0 0 3.5 0.5 0 0 1.8 0.3 21.4 25.9 0 0 7.3 0 0.1 1.5 0.3 0 0 0 2 0 2.3 0 0 0 0 2.8 0.1 0 4.2 0 0.5 1.6 11 0.9 0 0 1.5 1 1 6.9 10.7 0 0 0 44.3 14.2 0 0 10.7 0.3 0.5 0 0.1 0.1 1.1 0.8 0.4 0.3 1.7 0 0.1 0 0 0 0 0.9 3.3 4.5 0 0.6 0.2 0 0.2 0.5 0.9 0.1 0.6 0 1.2 0 0.1 2.1 0 0 0.3 0 0 0 6 3.3 5.4 24.3 0 0 0 0 0 0.5 0.3 0.8 0 0 0 0.3 0 0 0 0.3 0.6 2.8 1.8 0 0.8 3 0 0 0 10.1 0 0 0 0 0 23.7 0 0 0 0 0 5.7 31.3 0 2.9 0.3 0 0 9.7 0 0 3.9 0 0.1 0 0 0 0 0 7.2 3.4 4.7 0.3 0 0 0 0 0 0.1 0.8 14.7 0 0 0 0 0 0 0.1 0 0.2 0.2 Breese Pond Outlet 04 Breese Pond Outlet 03 Prindle Brook Fullam Brook Goodsell Brook Willow Brook Mallet's Creek Trib 8 Beaver Brook Sheldon Spring Trib 0.1 0.1 0 1.5 1.8 0.7 2.5 1.2 0 Urban vs Agricultural Urban vs Reference Agricultural vs Reference S p-value 0.11 0.00 0.08 D p-value 0.03 0.07 0.37 D S S 0 0 0 0 0 0 0 0 0 0.1 0.9 0 0 5.2 0 0 1.3 0 0 0 0 1.3 0 1.4 0 0 1 0 0 0 0 0 0 0 0.3 0 0 4.6 0 0 1.3 0 0.3 0 0 0.4 2.8 4.5 0.3 0 0 5 0.1 0.7 0.9 1.7 0 2 4.5 1.5 2.5 13.6 0 5 2.3 0.7 1.2 3.1 2.9 2 0.6 1.3 0.4 2.3 0.8 0 0.9 1.5 1.7 0 0.7 6.3 9.9 2.2 11 9.4 7.4 9.5 5.4 0 24.9 28.7 3.2 0 42.9 20.6 2.2 5 7.2 0 0 0 0 0 0 0 0 0 D D p-value 0.02 0.00 0.00 D Dip Parme sp 0 0 0 2 0 0.8 0.2 0.9 0 0 0 0 0 0 Col Sten cren 0 0 5.8 0 0 0 0 0 S 0 0 0 0 0 0 Dip Thiey grp 0 0 0 0 0.3 0 0 10 0 0 0 0 1.3 0 0 0 0 0 0 0 0 0 Tri Cheu sp 0 9 0 0 0 0 0 1.5 Rugg Brk Low Rock River Trout Brook Steven's Brook Wanzer Brk Rugg Brk High Saxe Brook Bump School Brook 0 0 0 0 0 0 Tri Hybr bett 0 3.6 0 0 0 15.9 Dip Simu tube Dip Cric sp 0 0 0 0 0 0 Eph Para sp Col Psep herr 0 0 0 0 0 12.6 Dip Tvet sp Tri Symp slos 0.4 0 0 3.4 14.9 0 Col Opti sp Ple Leuc imm 0 0 0 0.2 0 0 0 0 0 0.2 0 0 0 0 0.4 0.2 0 2.7 0 0 0 0 0 0 0 0 0 34.9 12.5 0.9 25.8 39.3 1.1 18.9 3.5 23.9 29.2 31.6 1.9 25.7 0 16 2.5 2.7 44.8 0.4 1.4 0 14 1.4 0.7 6 0 9.1 2.9 1.5 0.7 0.6 13.9 0 0 0 3.5 0.4 1.9 1.6 0.3 8.5 0 0.9 1.6 0 2.2 0.1 7.7 0.4 7.1 2.3 1.3 0 5.3 6 0.6 1.7 10.2 15.7 12.7 8.9 0 16.5 25 3 9 22 16.1 4.8 3.5 21.2 11.4 1.7 0.7 1.8 1.6 14.7 0 5.5 4 1.5 2.9 4.5 8.1 12.2 4.1 2 4.7 0.2 0 0.1 0.8 1.2 2.2 1.5 0.9 6.1 5.3 2.5 5.7 5.4 3.6 2.9 0.8 0.3 4.2 0.2 26 6.9 6.6 0.9 19.3 2 10 7.7 1.9 0.6 1.4 0.3 4.5 0 0 0 0 15.3 2 9.1 0 0 0.5 0.6 1.8 10.7 2.5 2.3 4.3 0 0.4 5.1 0 0 20.2 0.4 0.3 0.5 0.3 0 0.3 0.3 0 2 0 0 1.5 0.3 1.3 0 0.6 2.4 2.8 0 0.4 0.2 0 1.3 0.1 1.1 0.6 1 Eph Baet sp Ple Taenx sp 1.1 0 0 0 0 0 Dip Rheta dist Tri Chim ater 0 0 0 0 0 0.1 Dip Thiel sp Dip Poly avic 0.7 0.1 0 0 0 0 Dip Euki clar Dip Pseu sp 2.2 0 3.7 0 9.2 0 Dip Poly conv Tri Dipl sp 0 0 0 0 0.3 0 Eph Ephel sp Dip Micrs sp 0 0 0 0 0 0 Tri Dolo sp Oli Lumbd unid 0 0 0 0 0 0 Eph Stenm lute Col Ouli lati 0 0 0 0 0 0 Tri Rhya sp Dip Parcl sp 0 0 0 0 0 0 Ple Chlo imm Dip Hexe sp Urban Agricultural Reference no zeros is below this one Bartlett Brook Centennial Brook Englesby Brook Monroe Brook Morehouse Brook Potash Brook 0 1.2 0.4 0.9 0 0.3 0 0.4 0.7 3 35.5 0.3 0 0 1.5 0.4 0 0.4 0 0 0.4 0 0 0 0 0 0 2.1 0 0.1 0.2 0.4 0.3 0 0.9 8.6 0.2 4.3 0 0.4 1.5 0 0.6 2.2 0 2.4 0.1 0.4 0 0 0.3 0.3 0.3 8.7 0.8 0.6 8.4 0.2 15.8 3 1.9 2.6 2.4 7.5 1.2 0.4 0.3 0.8 7 2.4 0.1 0.1 0 0 0 0 0 0 0 0.3 0 0 0 2.2 0 0 0.2 0 0.3 0 0 0 0 0 0 5.3 0 1.4 0 3.6 0.8 2.7 0.8 0.8 1.1 1 6.6 3.2 2.2 0.3 0.9 9 1.1 0.5 1.1 S D p-value 0.06 0.02 0.01 D D D p-value 0.00 0.00 0.01 D S D D p-value 0.36 0.00 0.00 S: There is a not a significant difference in the % compositions of this group of taxa between the two groups of sites (between the two sectors). D: There is a significant difference in the % compositions of this group of taxa between the two groups of sites (between the two sectors).

Tri Rhya sp Eph Stenm lute Tri Dolo sp Eph Ephel sp 0 3.6 0 0 0 15.9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 9 0 0 0 0 0 1.5 0 0 0 0 0.3 0 0 10 0 0 5.8 0 0 0 0 0 0 0 0 2 0 0.8 0.2 0.9 0 0 0 0 0 0 0 4.1 0 0 0 0 0 0 0.2 2.6 0 0 0 0 3.5 0.5 0 0 1.8 0.3 21.4 25.9 0 0 7.3 0 0.1 1.5 0.3 0 0 0 2 0 S 2.3 0 0 0 0 2.8 0.1 0 4.2 0 0.5 1.6 11 0.9 0 0 1.5 1 1 6.9 10.7 0 0 0 44.3 14.2 0 0 10.7 0.3 0.5 0 0 0 0 0.1 0.9 0 0 5.2 0 0 1.3 0 0 0 0 1.3 0 1.4 0 0 1 0 0 0 0 0 0 0 0.3 0 0 4.6 D 0 0 1.3 0 0.3 0 0 0.4 2.8 4.5 0.3 0 0 5 0.1 0.7 0.1 0.1 1.1 0.8 0.4 0.3 1.7 0 0.1 0 0 0 0 0.9 3.3 4.5 0 0.6 0.2 0 0.2 0.5 0.9 0.1 0.6 0 1.2 0 0.1 2.1 0 0 0.3 0 0 0 6 3.3 5.4 24.3 0 0 0 0 0 0.5 0.3 0.8 0 0 0 0.3 0 0 0 0.3 0.6 2.8 1.8 0 0.8 3 0 0 0 10.1 0 0 0 0 0 23.7 0 0 0 0 0 5.7 31.3 0 2.9 0.3 0 0 9.7 0 0 3.9 0 0.1 0 0 0 0 0 7.2 3.4 4.7 0.3 0 0 0 0 0 0.1 0.8 14.7 0 0 0 0 0 0 0.1 0 0.2 0.2 0.9 1.7 0 2 4.5 1.5 2.5 13.6 0 5 2.3 0.7 1.2 3.1 2.9 2 0.6 1.3 0.4 2.3 0.8 0 0.9 1.5 1.7 0 0.7 6.3 9.9 2.2 11 9.4 7.4 9.5 5.4 0 24.9 28.7 3.2 0 42.9 20.6 2.2 5 7.2 0 0 0 0 0 0 0 0 0 Rugg Brk Low Rock River Trout Brook Steven's Brook Wanzer Brk Rugg Brk High Saxe Brook Bump School Brook 0 0 0 0 1.3 0 0 0 Breese Pond Outlet 04 Breese Pond Outlet 03 Prindle Brook Fullam Brook Goodsell Brook Willow Brook Mallet's Creek Trib 8 Beaver Brook Sheldon Spring Trib 0.1 0.1 0 1.5 1.8 0.7 2.5 1.2 0 Urban vs Agricultural Urban vs Reference Agricultural vs Reference S S p-value 0.11 0.00 0.08 D p-value 0.03 0.07 0.37 D S D D p-value 0.02 0.00 0.00 Dip Parme sp Ple Chlo imm 0 0 0 0 0 0 Col Sten cren Dip Cric sp 0 0 0 0 0 12.6 Dip Thiey grp Col Psep herr 0.4 0 0 3.4 14.9 0 Tri Cheu sp Tri Symp slos 0 0 0 0.2 0 0 Tri Hybr bett Ple Leuc imm 1.1 0 0 0 0 0 Dip Simu tube Ple Taenx sp 0 0 0 0 0 0.1 Eph Para sp Tri Chim ater 0.7 0.1 0 0 0 0 Dip Tvet sp Dip Poly avic 2.2 0 3.7 0 9.2 0 Col Opti sp Dip Pseu sp 0 0 0 0 0.3 0 0 0 0 0.2 0 0 0 0 0.4 0.2 0 2.7 0 0 0 0 0 0 0 0 0 34.9 12.5 0.9 25.8 39.3 1.1 18.9 3.5 23.9 29.2 31.6 1.9 25.7 0 16 2.5 2.7 44.8 0.4 1.4 0 14 1.4 0.7 6 0 9.1 2.9 1.5 0.7 0.6 13.9 0 0 0 3.5 0.4 1.9 1.6 0.3 8.5 0 0.9 1.6 0 2.2 0.1 7.7 0.4 7.1 2.3 1.3 0 5.3 6 0.6 1.7 10.2 15.7 12.7 8.9 0 16.5 25 3 9 22 16.1 4.8 3.5 21.2 11.4 1.7 0.7 1.8 1.6 14.7 0 5.5 4 1.5 2.9 4.5 8.1 12.2 4.1 2 4.7 0.2 D 0 0.1 0.8 1.2 2.2 1.5 0.9 6.1 5.3 2.5 5.7 5.4 3.6 2.9 0.8 0.3 4.2 0.2 26 6.9 6.6 0.9 19.3 2 10 7.7 1.9 0.6 1.4 0.3 4.5 0 0 0 0 15.3 2 9.1 0 0 0.5 0.6 1.8 10.7 2.5 2.3 4.3 0 0.4 5.1 0 0 20.2 0.4 0.3 0.5 0.3 0 0.3 0.3 0 2 0 0 1.5 0.3 1.3 0 0.6 2.4 2.8 0 0.4 0.2 0 1.3 0.1 1.1 0.6 1 Eph Baet sp Tri Dipl sp 0 0 0 0 0 0 Dip Rheta dist Dip Micrs sp 0 0 0 0 0 0 Dip Thiel sp Oli Lumbd unid 0 0 0 0 0 0 Dip Euki clar Col Ouli lati 0 0 0 0 0 0 Dip Poly conv Dip Parcl sp no zeros is below this one Bartlett Brook Centennial Brook Englesby Brook Monroe Brook Morehouse Brook Potash Brook Dip Hexe sp Reference Agricultural Urban Conclusion 0 1.2 0.4 0.9 0 0.3 0 0.4 0.7 3 35.5 0.3 0 0 1.5 0.4 0 0.4 0 0 0.4 0 0 0 0 0 0 2.1 0 0.1 0.2 0.4 0.3 0 0.9 8.6 0.2 4.3 0 0.4 1.5 0 0.6 2.2 0 2.4 0.1 0.4 0 0 0.3 0.3 0.3 8.7 0.8 0.6 8.4 0.2 15.8 3 1.9 2.6 2.4 7.5 1.2 0.4 0.3 0.8 7 2.4 0.1 0.1 0 0 0 0 0 0 0 0.3 0 0 0 2.2 0 0 0.2 0 0.3 0 0 0 0 0 0 5.3 0 1.4 0 3.6 0.8 2.7 0.8 0.8 1.1 1 6.6 3.2 2.2 0.3 0.9 9 1.1 0.5 1.1 S D D p-value 0.06 0.02 0.01 D D p-value 0.00 0.00 0.01 D S D p-value 0.36 0.00 0.00

Acknowledgements The authors wish to thank: The EPSCoR Baccalaureate College Summer Research Program under NSF Grant Number, EPS-0236976 The Norwich University Faculty Development Program for funding The Vermont DEC for permission to use their data in this presentation

Questions? For more information, go to: http://www2.norwich.edu/pinkhamc/