Document No. 12699-001-500 A-1 October 2011 

Appendix A 

Select photographs of different phases of the brown trout 

reproductive study 
Document No. 12699-001-500 A-1 December 2008 

Photo 1: Egg cups used for hatching of brown trout eggs; 10 replicates with 60 (target) eggs / replicate. 

Photo 2: Egg cup with eggs at test initiation. Photo includes egg pickers, container of remaining eggs for 

analytical, and counter. 
Document No. 12699-001-500 A-2 December 2008 

Photo 3: Eggs in egg cups for CC-350-008 brown trout eggs; 10 replicates with 60 eggs / replicate. 

Photo 4: Close up of eggs in egg cup at test initiation. 
Document No. 12699-001-500 A-3 December 2008 

Photo 5: Water bath covered with black curtain to keep brown trout eggs in the dark during initial stage. 

Photo 6: Test chambers in water bath for brown trout reproductive study during swim-up stage. 
Document No. 12699-001-500 A-4 December 2008 

Photo 7: Dilutor panel (one of two) used to feed adjusted Horsetooth water to each testing chamber. 

Photo 8: Separation of water bath into light conditions for swimup fry and dark for pre swimup yolk sac fry. 
Document No. 12699-001-500 A-5 December 2008 

Photo 9: Brown trout fry in test chamber at swim-up (prior to thinning). 

Photo 10: Brown trout fry in test chamber at swim-up (after thinning to 100 organisms). 
Document No. 12699-001-500 B-1 October 2011 

Appendix B 

Wet weight estimation for adult fish from Saratoga hatchery and 

raw data for adult fish 
Data from Brown trout Parental Study, 12699-001

filename: adult data.xls

Treatment

Total # 

eggs

Adult fish 

length 

(mm)

Adult fish 

weight (g) - 

CAS

Adult fish 

weight (g)

Predicted 

adult fish 

wt (g)

Se - wholebody 

(mg/kg 

dw)

 Se Egg 

(mg/kg 

dwt)

SC-001 4,173 498 1,393 1855.4 3.6 0.76

SC-002 4,005 420 826 1088.9 4.1 0.94

SC-003 5,120 520 1,553 2071.6 3.7 0.83

SC-004 1,248 562 2,500 3350.1 4.3 0.92

SC-005 5,448 558 2,187 2927.5 3 1.2

SC-006 3,176 439 842 1111.0 3.1 1.2

SC-007 3,224 449 1,175 1560.9 2.7 1

SC-008 4,005 494 1,446 1926.9 2.5 0.96

SPC-001 0.73

SPC-002 0.73

SPC-003 0.73

SPC-004 0.73

SPC-005 0.73

SPC-006 0.73

High

LSV2C-002 1,096 304 221 280 271.2 8.9 12.8

LSV2C-003 474 300 217 260 266.6 13.8 40.3

LSV2C-004 766 290 219 260 268.6 17.9 36

LSV2C-005 476 294 217 250 265.6 13.6 26.8

LSV2C-006 346 335 420 424.9 17.2 26.9

LSV2C-007 773 315 243 290 301.8 6.7 18.6

LSV2C-008 372 296 194 230 234.8 9.6 17.7

LSV2C-010 161 311 278 314 348.9 22.6 38.8

LSV2C-012 1,031 360 341 433.6 7.2 13.2

LSV2C-016 826 300 198 260 239.8 9.2 13.4

LSV2C-017 447 341 275 310 344.3 13.2 20.5

LSV2C-019 693 330 282 364 354.0 8.6 12.5

LSV2C-020 525 280 198 241 241.0 11.3 11.2

LSV2C-021 1,208 307 246 317 305.4 20 28.1

Bkd

CC-150-009 1,215 324 520 315 * 675.3 8.4 12.8

CC-150-011 488 342 303 351 382.4 5.6 8.4

CC-150-012 556 317 232 269 286.1 6.7 8.5

CC-150-013 1,234 332 283 376 355.1 5.9 8.4

CC-150-015 1,003 313 213 281 260.1 6 9.1

CC-150-016 1,658 391 468 621 604.7 7 7.5

CC-150-017 414 265 150 178 175.6 5.6 6.6

CC-150-018 959 308 224 279 275.0 4.7 6.9

CC-150-020 1,332 310 238 318 294.9 7.2 6.2

Low

CC-350-006 1,154 370 373 475 477.0 9.2 14

CC-350-007 1,174 350 321 416 407.3 5.5 6.9

CC-350-008 922 335 254 341 315.7 8.5 9.5

min (field) 161 265 150 178 176 4.7 6.2

max (field) 1658 391 520 621 675 22.6 40.3

* value not included in developing wet weight estimates for SC hatchery fish
Relationship between wet wt data of field fish from CAS and from field measurem

y = 1.351x - 27.062

R

2 = 0.963

0

100200300400500600700

0 100 200 300 400 500 

wt (CAS)

wt (field)

Note: dashed line of 1.0
Document No. 12699-001-500 C-1 October 2011 

Appendix C 

Egg counts for Saratoga hatchery fish (SC) and field collected fish 
Estimation of Brown trout eggs numeration using volume-estimating technique, 12699-001

October 25, 2007

Hatchery Eggs from Saratoga National Fish Hatchery (SNFH, Saratoga, WY)

The number of eggs placed into each study was counted manually. For all of the treatments, the remaining

number of eggs was estimated using a volume technique to develop a #egg/volume ratio for that particular female.

The technique consisted of counting the number of eggs that filled a graduated cylinder to a particular volume

(e.g., 50 mL) to determine the # of eggs per mL for that female. Initially we conducted two separate counts by 

two different staff personnel. Based on the agreement of these counted numbers for the particular volume, we

subsequently only conducted counts once per female. Using this ratio, we then calculated the total number of

remaining eggs for the total volume of eggs measured in a graduated cylinder. The total number of eggs used to

initiate the studies (i.e., 600) was then added to the estimated number of remaining eggs to determine the total

number of eggs for that particular female trout.

# Eggs placed # eggs counted Avg # eggs Vol of eggs Avg # Total vol (ml) Total

Treatment in study Count# to est. #eggs/vol counted counted (ml) eggs/mL of remaining eggs # eggs

SC-001 600 1 1058 1051 100 10.51 340 4,173

2 1044 100

SC-002 600 1 570 567.5 50 11.35 300 4,005

2 565 50

SC-003 600 1 452 452 50 9.04 500 5,120

SC-004 600 1 506 506 50 10.12 64 1,248

SC-005 600 1 480 480 50 9.6 505 5,448

SC-006 600 1 477 477 50 9.54 270 3,176

SC-007 600 1 495 495 50 9.9 265 3,224

SC-008 600 1 532 532 50 10.64 320 4,005

Avg 3,800

Geomean 3,525

SD 1,305

CV 34%
Estimation of Brown trout eggs numeration using direct counts and volume-estimation technique, 12699-001

November 16 & 17, 2007

Treatment eggs from background, low, and high treatment areas (ID)

The number of eggs placed into each study was counted manually. For almost all of the treatments, the 

remaining eggs were also counted manually. When there were too many eggs to count, the number of

remaining eggs were estimated using a volume technique to develop a #eggs/volume ratio for that particular 

female. The technique consisted of counting the number of eggs that fill a graduated cylinder to a particular 

volume (e.g., 50 mL) to determine the # eggs per mL for that female. Using this ratio, we then calculated the

total number of remaining eggs for the total volume of eggs measured in the graduated cylinder. The total

number of eggs used to initiate the studies (e.g., 600) was then added to this estimated number of remaining

eggs to determine the total number of eggs for that particular female trout.

# Eggs placed # Remaining # eggs counted Vol of eggs Avg # Total vol (ml) Total

Treatment in study eggs counted to est. #eggs/vol counted (ml) eggs/mL of remaining eggs # eggs

High

LSV2C-002 600 496 1,096

LSV2C-003 400 74 474

LSV2C-004 500 266 766

LSV2C-005 300 176 476

LSV2C-006 eggs dead / dying upon arrival (not counted)

LSV2C-007 500 273 773

LSV2C-008 300 72 372

LSV2C-010 100 61 161

LSV2C-012 600 431 1,031

LSV2C-016 600 226 826

LSV2C-017 300 147 447

LSV2C-019 500 193 693

LSV2C-020 400 125 525 Avg

LSV2C-021 600 608 1,208 681

Bkd

CC-150-009 600 615 1,215

CC-150-011 300 188 488

CC-150-012 350 206 556

CC-150-013 600 634 1,234

CC-150-015 600 403 1,003

CC-150-016 600 460 50 9.2 115 1,658

CC-150-017 250 164 414

CC-150-018 600 359 959 Avg

CC-150-020 600 732 1,332 984

Low

CC-350-006 600 554 1,154

CC-350-007 600 574 1,174 Avg

CC-350-008 600 322 922 1,083








Document No. 12699-001-500 D-1 October 2011 

Appendix D 

Summary of fungal treatment methods for SC Hatchery eggs and 

field collected eggs 
1 

Treatment of brown trout eggs for fungal control. Study # 12699-001 

Different methods were employed in order to help control fungal growth and improve overall 

success rate of the brown trout eggs in the reproductive study. As mentioned in the report, 

this primarily affected only the SC hatchery eggs because they were the first batch to get in. 

And additional methods were employed to reduce fungal growth with the field collected eggs 

(initiated Nov. 16 & 17, 2008) and the second set of hatchery eggs (initiated Dec. 4, 2007). 

November 2, 2007: 

1% salt (as NaCl) solution was added to SC-002 and SC-004 chambers (i.e., 50 g of NaCl 

was mixed with small amount of water from each test chamber until all salt was dissolved 

and added). Measured chloride concentration at 5,620 mg/L. Time = 1700 hrs. 

Conductivity check at 1920 hrs: 

SC-004 @ 12,800 μS/cm2

SC-002 @ 11,440 μS/cm2 

Background conductivity @ 123 μS/cm2

November 3, 2007: 

Conductivity check in chambers @ 0550 hrs 

SC-004 @ 1,770 μS/cm2

SC-002 @ 2,670 μS/cm2 

Background conductivity @ 118 μS/cm2

1% salt (as NaCl) solution was added to SC-003 (@0630 hrs) and SC-001 (@0700 hrs) 

chambers (i.e., 50 g of NaCl was mixed with small amount of water from each test chamber 

until all salt was dissolved and added). 

Conductivity check in chambers @ 1620 hrs 

SC-004 @ 343 μS/cm2

SC-002 @ 602 μS/cm2 

SC-001 @ 3,660 μS/cm2

SC-003 @ 3,830 μS/cm2

November 6, 2007: 

Installed small UV disinfection system on HT water line prior to going into 5-gallon head tank 

bucket above dilutor panel. This treats all water going into the study. Because there was 

algal / fungal growth in 5-gallon head tank, treated head tank with NaCl (189 g mixed in 5 

gallons). Salt solution was allowed to flush through the dilutor and into the test chambers 

(SC test chambers only). 

November 9, 2007: 

Added 50 g of NaCl to each SC chamber because still seeing fungus (Saprolegnia?). 

November 12, 2007: 

Now that eggs were eyed up, salt treatment consisted of removing egg cups from test 

chamber (SC treatments) and soaking in 3% salt treatment for 1 hour, and then transferring 
2 

back to cleaned test chamber. Treated drip lines with (~1,700 ppm) formalin. The formalin 

solution did not go into test chambers with eggs. 

November 16, 2007: 

A 1.5% NaCl solution (75 g NaCl) was added to 5 L water in test container for SC hatchery 

eggs. 

November 24, 2007: 

Saw some dead eggs in newly received field egg treatments. Add 190 g of NaCl to 5 gal 

head tank. Let it flush through dilutor and into all chambers. 

 November 25, 2007: 

Measured conductivity @ 0800 hrs, ~200 μS/cm2

. Moved all SC hatchery eggs into new egg 

cups. Let soak in 3% NaCl solution (60 g / 2 L) for ~ 1 h while being transferred into new egg 

cups. Some eggs were very covered by fungus. SC005 treatment appeared to have a lot of 

unfertilized (non-eyed up) eggs. Cleaned (soap and water wash) SC test chambers while 

eggs were in NaCl soak (all SC treatments). 

Treated all test chambers in bath #2 (ENSR water bath #3) with 1.5% NaCl, except SC 

chambers. Mixed 75 g NaCl with deionized water (Milli-Q) and poured into each chamber. 

15 test chambers in all for bath #2. 

Consisted of: CC-350-(LOW)-007 

 CC-150-(BKD)-009, -012, -015, & -016 

 LSV2C-(HIGH)-004, -005, -007, -008, -017, -019, & -020 

 SPC-(2ND HATCHERY)-002, -003, & -006 

November 26, 2007: 

Treated all test chambers in bath #1 (ENSR water bath #4) with 1.5% NaCl, except SC 

chambers. Mixed 75 g NaCl with deionized water (Milli-Q) and poured into each chamber. 

16 test chambers in all for bath #1. 

Consisted of: CC-350-(LOW)-006 & -008 

 CC-150-(BKD)-011, -013, -017, -018, & -020 

 LSV2C-(HIGH)-002, -003, -010, -012, -016, & -021 

 SPC-(2ND HATCHERY)-001, -004, & -005 

Drained and cleaned large circular (trout tank) hardness mixing tank. Chambers were static 

for ~2 hours. 

November 28, 2007: 

Soaked SC treatments by placing egg cups in 3% NaCl for one hour (120 g / 4 L) in separate 

chamber. Removed and placed back into test chambers. Cleaned as necessary. 
3 

November 29, 2007: 

After discussions with S. Covington and hatchery personnel, decided to use formalin to help 

minimize fungal growth on SC eggs. Treated SC-001 eggs with formalin (~1700 ppm, 6.8 ml 

in 4 L), by placing egg cup in separate chamber with formalin (next to original test chamber in 

water bath). From 1054 hrs to 1115 hrs. 

Also treated with following test chambers with formalin in a similar way. 

 SC-002 (1124 hrs to 1140 hrs) 

 SC-003 (1142 hrs to 1157 hrs) 

LSV2C-(HIGH)-007 (1720 hrs to 1738 hrs). 

Noticed a lot of dead / dying eggs in the LSV2C-(HIGH)-007 treatment at the end of the day 

so decided to treat with formalin, even though it did not appear that there was fungus on 

these eggs1. 

November 30, 2007: 

Treated the following egg treatments with formalin (~1700 ppm, as 7 ml in 4 L). Performed in 

secondary chamber as before, soaked for ~30 min, and returned to original chamber. 

Started soak at 1520 hrs, finish at 1550 hrs. 

 SC-001, SC-005, SC-007, SC-008 (bath #1) 

 SC-004, SC-002, SC-003, SC-006 (bath #2) 

December 1, 2007: 

Placed egg cup for SC-001 in old formalin solution (prepared on Nov. 30, but still in water 

bath) for 15 minutes. Returned to original test chamber. 

December 2, 2007: 

Treat SC-001 with formalin by mixing 8.5 ml in 200 ml water (i.e., cold hardness adjusted 

Horsetooth water, similar as all preps before). Poured this mixture directly into test chamber 

(assumes ~5L so target is ~1700 ppm). After 15 min, pur 4 L HT water into test chamber to 

flush out formalin. 

Still seeing fungus on control eggs, try a 4% NaCl solution on 1 SC batch for 1 hour. 

Prepared by dissolving 200 g NaCl in 1 L of water and pouring directly into test chamber 

(assumes 5 L of water in chamber). Added to SC-005 @ 1340 hrs. 

Treated the following test chambers with formalin (8.5 ml into test chamber for 15 minutes, 

followed by 4-5L flush): SC-004, SC-002, SC-006 

December 3, 2007: 

Observed first hatched out alevin in SC-004 and 1 egg that appeared to explode (termed 

‘dead while hatching’). 

 

1 None of these eggs appeared to be fertilized and all eventually died. 
4 

No more treatments for SC test chambers. 

December 5, 2007: 

Added 1% NaCl (50 g NaCl, assuming 5 L water in each chamber) to all test chambers 

except SC treatments (-001 through -008), and SPC-002, and SPC-004 (small SPC test 

chambers, i.e., ~ 2.5 L chambers). 

December 7, 2007: 

Clean out test chambers for LSV2C-008 and -019 with soap and water and replaced in bath. 

December 8, 2007: 

Cleaned egg cups by brushing outside surface with toothbrush. Did not disturb eggs. After 

cleaning egg cup, placed egg cup into a cleaned test chamber. This was done for the 

following treatments. 

LSV2C-(HIGH)-017, -020, -007 (transferred few remaining eggs to a new egg cup), 

-005, -004, -010, -002, & -016 

December 9, 2007: 

Cleaned egg cups by brushing outside surface with toothbrush. Did not disturb eggs. After 

cleaning egg cup, placed egg cup into a cleaned test chamber. Also replaced aeration 

pipettes. This was done for the following treatments. 

LSV2C-(HIGH)-021, -012, & -003 

December 10, 2007: 

Cleaned egg cups by brushing outside surface with toothbrush. Did not disturb eggs. After 

cleaning egg cup, placed egg cup into a cleaned test chamber. Also replaced aeration 

pipettes. This was done for the following treatments. 

CC-350-(LOW)-006, -007, & -008 

CC-150-(BKD)-009, -015, & -016 

December 7 – 18, 2007: 

Noticed that flows on some treatments were reduced due to fouling of tubing lines (from 

splitter box to test chamber). Flushed out lines with hydrogen peroxide (1.5%). Test 

chambers were static during the flush out and either overnight or during the day (~ 8 hr) while 

water was allowed to flush through the tubing to waste. Also blew biofilm out of some lines, 

but this did not work for all chambers. This was done over a period of time because there 

were only a few extra tubing lines that did not directly feed a treatment. Therefore, a few 

were cleaned each day and allowed to flush prior to using for a test chamber. 

December 31, 2007: 

Egg cup for LSV2C-(HIGH)-019 was very dirty so changed out with clean egg cup. 
5 

January 1, 2008: 

Egg cups for the following treatments were very dirty so changed out with clean egg cups: 

 LSV2C-(HIGH)-021, -004 & -012 

 CC-150-(BKD)-013 & -012 

 CC-350-(LOW)-006 

January 2, 2008: 

Cleaned dilutor panels (scrubbing only) to remove biofilm buildup. Test chambers were 

static for ~ 1 hr during this time. 

January 18, 2008: 

Exchanged test chamber for LSV2C-(HIGH)-003 with clean one due to fungal growth and 

high mortality. 
Document No. 12699-001-500 E-1 October 2011 

Appendix E 

Summary of water quality data selenium and sulfate analysis in 

water and selenium concentrations in eggs measure during 

reproductive study 




















Average Daily Chamber temp

0.02.04.06.08.0

10.012.014.0

5-Oct-07 25-Oct-07 14-Nov-07 4-Dec-07 24-Dec-07 13-Jan-08 2-Feb-08 

Time

Avg chamber temp (C) Bath 3 Bath 4
Water temperature in test chambers for Brown trout studies

12699-001

filename: water qual.xls

SC treatments SPC treatments CC-150 (BKD) treatments CC-350 (LOW) treatments LSV2C (HIGH) treatments

Water temperature (°C) Water temperature (°C) Water temperature (°C) Water temperature (°C) Water temperature (°C)

Water Water Water Water Water Water Water Water Water Water Daily Avg Temp (°C)

Date Test Day bath #3 bath #4 Test Day bath #3 bath #4 Test Day bath #3 bath #4 Test Day bath #3 bath #4 Test Day bath #3 bath #4 bath #3 bath #4

25-Oct-07 0

26-Oct-07 1

27-Oct-07 2 11.3 9.9 11.3 9.9

28-Oct-07 3 10.5 9.1 10.5 9.1

29-Oct-07 4 11.3 9.8 11.3 9.8

30-Oct-07 5 11.9 10.0 11.9 10.0

31-Oct-07 6 10.8 9.3 10.8 9.3

1-Nov-07 7 9.5 10.4 9.5 10.4

2-Nov-07 8 9.1 9.8 9.1 9.8

3-Nov-07 9 9.3 9.1 9.3 9.1

4-Nov-07 10 10.3 10.0 10.3 10.0

5-Nov-07 11 10.3 9.7 10.3 9.7

6-Nov-07 12 9.3 9.1 9.3 9.1

7-Nov-07 13 10.2 9.7 10.2 9.7

8-Nov-07 14 10.0 9.5 10.0 9.5

9-Nov-07 15 11.2 10.9 11.2 10.9

10-Nov-07 16 10.0 9.5 10.0 9.5

11-Nov-07 17 9.7 9.5 9.7 9.5

12-Nov-07 18 9.8 9.7 9.8 9.7

13-Nov-07 19 9.7 9.7 9.7 9.7

14-Nov-07 20 8.9 9.1 8.9 9.1

15-Nov-07 21 8.5 9.0 8.5 9.0

16-Nov-07 22 10.3 9.6 0 10.3 9.6 10.3 9.6

17-Nov-07 23 9.7 9.5 0 0 1 9.7 9.5

18-Nov-07 24 9.8 9.6 1 1 2 9.8 9.6

19-Nov-07 25 10.6 9.7 2 10.3 10.3 2 10.8 10.6 3 10.5 9.7 10.6 10.1

20-Nov-07 26 10.2 9.7 3 10.3 9.2 3 9.9 10.3 4 10.1 10.2 10.1 9.9

21-Nov-07 27 9.4 9.1 4 9.7 9.5 4 9.9 10.2 5 9.7 9.3 9.7 9.5

22-Nov-07 28 9.3 9.4 5 9.4 10.1 5 9.6 9.4 6 9.6 10.4 9.5 9.8

23-Nov-07 29 9.8 9.3 6 9 9.8 6 9.8 10.3 7 9.3 8.8 9.5 9.6

24-Nov-07 30 9.7 9.3 7 9.1 9.6 7 9.8 10.4 8 9.8 10.6 9.6 10.0

25-Nov-07 31 10.0 9.3 8 10.7 10.2 8 10.4 9.6 9 10.5 10.2 10.4 9.8

26-Nov-07 32 9.8 9.6 9 9.8 9.5 9 10.1 10.4 10 10.4 10.1 10.0 9.9

27-Nov-07 33 9.7 9.3 10 9.2 9.4 10 9.6 9.3 11 10.2 10 9.7 9.5

28-Nov-07 34 10.3 9.6 11 9.8 10.1 11 10.1 10.5 12 9.8 9.4 10.0 9.9

29-Nov-07 35 10.1 9.5 12 9.9 9.7 12 10.3 10.4 13 10.4 9.5 10.2 9.8

30-Nov-07 36 9.2 9.4 13 10.3 9.5 13 9.3 9.7 14 9.8 9.8 9.7 9.6

1-Dec-07 37 9.4 8.7 14 8.7 9.8 14 9.1 9.5 15 9.1 9.2 9.1 9.3

2-Dec-07 38 9.1 9.3 15 8.8 9.1 15 9.5 10.1 16 9.6 10.4 9.3 9.7

3-Dec-07 39 9.9 9.6 16 10.4 9.9 16 10 9.4 17 9.9 9.4 10.1 9.6

4-Dec-07 40 11.2 10.2 0 11.3 10.1 17 10.3 10.4 17 10.8 10.1 18 11 11.2 10.9 10.4

5-Dec-07 41 10.2 9.4 1 10.2 10.1 18 10.7 9.5 18 10.4 10.3 19 10.4 10.1 10.4 9.9

6-Dec-07 42 9.6 9.7 2 9.4 9.4 19 9.8 9.3 19 10.1 9.5 20 10.1 9.3 9.8 9.4

7-Dec-07 43 10.0 9.3 3 10.5 9.3 20 9.7 9.3 20 10.3 10.1 21 9.5 9.2 10.0 9.4

8-Dec-07 44 10.1 9.8 4 9.3 10 21 9.1 9.8 21 9.8 9.6 22 9.8 10.1 9.6 9.9

9-Dec-07 45 9.7 9.4 5 9.3 9.7 22 9.7 10.1 22 9.7 9.8 23 9.6 9.8 9.6 9.8

10-Dec-07 46 9.7 9.3 6 9.8 9.5 23 9.9 9.5 23 9.6 9.4 24 9.1 9.4 9.6 9.4

11-Dec-07 47 9.8 9.4 7 9.2 9.3 24 8.9 9.3 24 9.1 9.8 25 9.3 10.1 9.3 9.6

12-Dec-07 48 8.5 9.1 8 8.6 9 25 8.9 9.5 25 8.8 9.1 26 8.7 9.1 8.7 9.2

13-Dec-07 49 10.0 9.6 9 10 9.7 26 10.5 9.9 26 10.4 9.5 27 10.4 9.6 10.3 9.7

14-Dec-07 50 10.7 10.0 10 10 10.2 27 10.4 10.1 27 10.3 11.1 28 10.7 10.5 10.4 10.4

15-Dec-07 51 10.5 10.3 11 9.8 10 28 9.7 10.1 28 10.4 10 29 10.5 10.2 10.2 10.1

16-Dec-07 52 10.1 10.5 12 10.7 10.1 29 10.3 10 29 10.4 11.3 30 10.5 9.8 10.4 10.3

17-Dec-07 53 11.3 10.4 13 10.8 10.7 30 11.3 11.1 30 11 10.4 31 10.9 12.1 11.1 10.9

18-Dec-07 54 11.4 10.1 14 10.2 10.3 31 10.7 10.3 31 11 11.5 32 11 10.9 10.9 10.6

19-Dec-07 55 11.8 10.2 15 11.2 10.3 32 9.7 10.3 32 11.1 10.9 33 10.7 10.9 10.9 10.5

20-Dec-07 56 11.3 10.6 16 10.9 10.6 33 11.3 11.2 33 11.3 10.4 34 10.7 10.2 11.1 10.6

21-Dec-07 57 10.6 10.6 17 10.1 10.4 34 11.2 10.6 34 10.9 11.4 35 10.6 10.4 10.7 10.7

22-Dec-07 58 10.3 10.5 18 10.1 11 35 10.9 10.6 35 10.9 11 36 10.1 9.9 10.5 10.6

23-Dec-07 59 10.0 10.5 19 11.1 10.3 36 10.9 10 36 10.9 10.1 37 10.7 10.3 10.7 10.2

24-Dec-07 60 10.9 10.2 20 10.6 10.4 37 10.9 11.2 37 10.9 11.5 38 10.7 11.4 10.8 10.9

25-Dec-07 61 10.2 10.1 21 10.9 10.7 38 10.9 10.1 38 10.8 10.2 39 10.2 10.1 10.6 10.2

26-Dec-07 62 22 39 39 40

27-Dec-07 63 9.8 10.5 23 10.4 10.3 40 10.2 10.5 40 10.3 11.2 41 10.7 10.9 10.3 10.7

28-Dec-07 64 9.7 10.4 24 9.7 9.9 41 10.2 10.3 41 9.9 9.7 42 10 9.9 9.9 10.0

29-Dec-07 65 10.7 10.7 25 9.5 10.4 42 9.9 10.4 42 10.1 10.8 43 10 10.7 10.0 10.6

30-Dec-07 66 10.5 11.1 26 10.4 10 43 10.2 10.5 43 10.3 10.2 44 9.9 10.7 10.3 10.5

31-Dec-07 67 10.5 10.8 27 9.9 11.1 44 10.3 9.6 44 10.5 11 45 10.3 9.8 10.3 10.5

1-Jan-08 68 11.1 10.8 28 10.3 10.5 45 10.3 9.7 45 10.1 10.1 46 9.5 10.5 10.3 10.3

2-Jan-08 69 11.1 10.5 29 9.8 9.8 46 9.9 9.9 46 10.2 10.7 47 9.7 10.4 10.1 10.3

3-Jan-08 70 30 47 47 48

4-Jan-08 71 10.7 10.6 31 11.1 10.6 48 10.6 10.5 48 10.5 9.8 49 10.5 10.4 10.7 10.4

5-Jan-08 72 10.9 11.0 32 10.9 10.6 49 10.3 10.4 49 10.2 10 50 10.6 10.6 10.6 10.5

6-Jan-08 73 11.1 10.6 33 10.8 10.9 50 10.9 10.4 50 10.3 9.8 51 10.4 9.8 10.7 10.3

7-Jan-08 74 10.8 10.5 34 10.6 10.5 51 10 9.7 51 10.2 9.9 52 10.3 10.3 10.4 10.2

8-Jan-08 75 10.7 10.9 35 10.9 10.6 52 10.3 9.5 52 10.2 9.9 53 9.5 10.3 10.3 10.2

9-Jan-08 76 11.1 11.2 36 11.1 11.3 53 10.2 11 53 10.3 10.1 54 10.7 10 10.7 10.7

10-Jan-08 77 11.4 11.1 37 11.1 10.9 54 10.7 10.5 54 10.7 10.3 55 10.1 10.3 10.8 10.6

11-Jan-08 78 11.3 11.3 38 11.4 11.1 55 10.6 10.6 55 10.8 11.1 56 10.9 11.2 11.0 11.1

12-Jan-08 79 10.9 11.4 39 11.3 11.4 56 10.1 11 56 10.5 10.2 57 10.5 10.2 10.7 10.8

13-Jan-08 80 11.2 11.3 40 11.3 10.7 57 10.3 10.7 57 10.5 10.3 58 10.6 10.9 10.8 10.8

14-Jan-08 81 10.1 10.9 41 9.9 10 58 9.9 10.2 58 9.7 10 59 10.1 10.5 9.9 10.3

15-Jan-08 82 11.3 10.9 42 11.3 11.1 59 10.9 10.3 59 10.7 10.3 60 10.9 10.7 11.0 10.7

16-Jan-08 83 11.4 11.1 43 11.6 11.6 60 11.4 10.7 60 11.3 11.3 61 11.3 10.9 11.4 11.1

17-Jan-08 84 10.8 11.1 44 11 10.7 61 10.3 10 61 10.8 10 62 10.1 9.7 10.6 10.3

18-Jan-08 45 11.7 11.6 62 10.5 11.7 62 11.5 11.4 63 11.1 10.5 11.2 11.3

19-Jan-08 46 11 10.6 63 11 10.7 63 11 10.1 64 10.5 10.1 10.9 10.4

20-Jan-08 47 11.5 11.9 64 11.7 11.8 64 11.5 11.7 65 11.5 11.7 11.6 11.8

21-Jan-08 48 11.2 10.7 65 10.4 10.3 65 11.1 10.7 66 10.9 10.9 10.9 10.7

22-Jan-08 49 10.5 10.3 66 10.3 10.3 66 10.4 10.2 67 10.7 9.8 10.5 10.2

23-Jan-08 67 11.1 11.2 67 11.7 11.4 68 11.1 11.9 11.3 11.5

24-Jan-08 68 11.6 11.4 68 11.7 11.7 69 11.6 10.7 11.6 11.3

25-Jan-08 69 10.6 10.7 69 11.5 11.1 70 11.3 11.4 11.1 11.1

26-Jan-08 70 11 11.5 70 11.6 11.9 71 11.2 11.1 11.3 11.5

27-Jan-08 71 10.9 11.2 71 11.2 11 72 11.8 11.7 11.3 11.3

28-Jan-08 72 11.5 12.4 72 12.2 11.6 73 11.5 12.4 11.7 12.1

29-Jan-08 73 11.3 10.6 73 11.4 11.6 74 10.9 10.1 11.2 10.8

30-Jan-08 74 11 11.1 74 11.8 11.3 75 11.5 11.9 11.4 11.4

31-Jan-08 75 11.2 11.3 75 12 12 76 11.9 12.3 11.7 11.9

1-Feb-08 76 11.8 11.6 76 12.1 11.9 77 11.4 12.1 11.8 11.9

2-Feb-08 77 10.9 11.2 77 11.5 11.7 78 10.7 11.3 11.0 11.4

3-Feb-08 78 11 11.1 78 11.4 11.4 79 10.9 11 11.1 11.2

4-Feb-08 79 11.8 11.1 79 12.2 12.1 80 11.8 10.8 11.9 11.3

5-Feb-08 80 11.5 11.2 80 11.5 11.3 81 10.7 11.7 11.2 11.4

6-Feb-08 81 11.3 11.7 81 11.7 11.3 82 11.5 11.7 11.5 11.6

7-Feb-08 82 12.1 11.7 82 12.1 83 12.1 11.8 12.1 11.9

8-Feb-08 83 11.8 11.1 83 11.6 84 12.1 11.9 12.0 11.5

9-Feb-08 84 12.5 12.3 84 11.8 85 12.5 11.7 12.5 11.9

10-Feb-08 85 11.4 10.9 85 11.5 86 11.8 12.1 11.6 11.5

11-Feb-08 86 12.1 11.5 86 11.8 87 12.2 11.7 12.2 11.7

12-Feb-08 87 11.9 11.9 87 11.4 88 12 11.7 12.0 11.7

Avg 10.1 10.5 10.5 10.6 10.6

Std 0.733 0.680 0.798 0.782 0.822

Min 8.5 8.6 8.7 8.8 8.7

Max 11.9 11.9 12.5 12.2 12.5
Water hardness measured in dilution water during BT study

0

10203040506070

5-Oct-07 25-Oct-07 14-Nov-07 4-Dec-07 24-Dec-07 13-Jan-08 2-Fe

Time

Hardness (mg/L as CaCO3)
4 of 5

Water hardness (mg/L) measurements in Brown trout study (12699-001)

filename: water qual.xls

Water hardness

Date Test Day (mg/L)

25-Oct-07 D0

26-Oct-07 D1

27-Oct-07 D2

28-Oct-07 D3

29-Oct-07 D4 50

30-Oct-07 D5 54

31-Oct-07 D6 50

1-Nov-07 D7 48

2-Nov-07 D8 52

3-Nov-07 D9 54

4-Nov-07 D10 48

5-Nov-07 D11 58

6-Nov-07 D12 50

7-Nov-07 D13 50

8-Nov-07 D14 50

9-Nov-07 D15 46

10-Nov-07 D16 48

11-Nov-07 D17 54

12-Nov-07 D18 52

13-Nov-07 D19 48

14-Nov-07 D20 52

15-Nov-07 D21 48

16-Nov-07 D22 50

17-Nov-07 D23 48

18-Nov-07 D24 54

19-Nov-07 D25 54

20-Nov-07 D26

21-Nov-07 D27 46

22-Nov-07 D28 50

23-Nov-07 D29 50

24-Nov-07 D30 46

25-Nov-07 D31 48

26-Nov-07 D32 50

27-Nov-07 D33 50

28-Nov-07 D34 46

29-Nov-07 D35 50

30-Nov-07 D36 46

1-Dec-07 D37 48

2-Dec-07 D38 50

3-Dec-07 D39

4-Dec-07 D40 50

5-Dec-07 D41 54

6-Dec-07 D42 50

7-Dec-07 D43 48

8-Dec-07 D44 50

9-Dec-07 D45 48

10-Dec-07 D46 46

11-Dec-07 D47 52

12-Dec-07 D48 54

13-Dec-07 D49 50

14-Dec-07 D50 50

15-Dec-07 D51 48

16-Dec-07 D52 50

17-Dec-07 D53 48

18-Dec-07 D54 48

19-Dec-07 D55 50

20-Dec-07 D56 46

21-Dec-07 D57 50

22-Dec-07 D58 50

23-Dec-07 D59 52

24-Dec-07 D60 54

25-Dec-07 D61 52
5 of 5

Water hardness (mg/L) measurements in Brown trout study (12699-001)

filename: water qual.xls

Water hardness

Date Test Day (mg/L)

26-Dec-07 D62

27-Dec-07 D63 50

28-Dec-07 D64 50

29-Dec-07 D65 54

30-Dec-07 D66 50

31-Dec-07 D67 48

1-Jan-08 D68 50

2-Jan-08 D69 52

3-Jan-08 D70 50

4-Jan-08 D71 44

5-Jan-08 D72 42

6-Jan-08 D73 54

7-Jan-08 D74 48

8-Jan-08 D75 48

9-Jan-08 D76 50

10-Jan-08 D77 44

11-Jan-08 D78 44

12-Jan-08 D79 44

13-Jan-08 D80 44

14-Jan-08 D81

15-Jan-08 D82 40 SC fish (Oct. 25 - Jan. 17)

16-Jan-08 D83 44 Avg StDev Min Max

17-Jan-08 D84 50 49.32 3.19 40 58

18-Jan-08 44

19-Jan-08 42

20-Jan-08 44 SPC fish (Dec. 4 - Jan. 22)

21-Jan-08 52 Avg StDev Min Max

22-Jan-08 52 48.63 3.55 40 54

23-Jan-08 46

24-Jan-08 54

25-Jan-08 50

26-Jan-08 44

27-Jan-08 38

28-Jan-08 48

29-Jan-08 44

30-Jan-08 48

31-Jan-08 46

1-Feb-08 48

2-Feb-08 42

3-Feb-08 48

4-Feb-08 46

5-Feb-08 48

6-Feb-08 46

7-Feb-08 54

8-Feb-08 50

9-Feb-08 52

10-Feb-08 48 CC-150 & CC-350 fish (Nov. 17- Feb. 12)

11-Feb-08 Avg StDev Min Max

12-Feb-08 44 48.36 3.51 38 54

Average (overall) 48.78

StDev 3.506 LSV2C fish (Nov. 16- Feb. 12)

Min 38 Avg StDev Min Max

Max 58 48.38 3.49 38 54