NC0024112_Report_20041209

DMSION OF WATER QUALITY
December 1, 2004
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To: Dave Goodrich T
Through: Matt Matthews t`4 A" D
From: Kevin Bowden M D
DEC 9 2004
Subject: Toxicity Identification Evaluation
DWQ's Copper and Zinc Action Level Policy DENR - WATER OUAUTY
City of Thomasville - Hamby Creek W WTP Po1NT SOURCE BRANCH
NPDES No. NC0024112
Davidson County ,,, ...,..,�,.p,�,,:,,-,..w.�w.•'
Our office received Toxicity Identification Evaluation (TIE) information from the City of Thomasville
concerning their efforts to rule out copper and zinc as causative effluent toxicants. The information package was
transmitted by cover letter to our office from Mr. Morgan Huffman, Wastewater Treatment Plant Superintendent and was
received on 11/18/04. In our opinion it can't be determined whether copper and/or zinc caused the facility's WET
failures in November and December of 2003. No WET failures occurred subsequently and no meaningful TIE work
could be conducted. The facility remains subject to the copper/zinc policy should WET failures occur in the future.
Three distinct TIE testing events occurred during the nine month period the facility was under DWQ's Copper
and Zinc Action Level Policy. Initial effluent samples for TIE testing were collected on 3/29/04, 6/7/04 and 8/23/04. For
each series, seven (7) treatments were performed followed by eight (8) treatments "spiked " with 20 ug/L copper and 60
ug/L zinc. Effluent testing via "spiked" treatments was initiated at a later time with the initial sample. All treatments
were conducted with 100% effluent. Mean control organism/treatment reproduction, percent reduction and
corresponding copper/zinc initial sample concentrations are listed below.
Treatment (test initiated 3/31/04)
sample collected 3/29/04
Control
Treatment 1 (100% effluent, baseline)
Treatment 2 (0.5 mg/L EDTA)
Treatment 3 (3.0 mg/L EDTA)
Treatment 4 (8.0 mg/L EDTA)
Treatment 5 (0.5 mg/L Na2SO4)
Treatment 6 (3.0 mg/L Na2SO4)
Treatment 7 (8.0 mg/L Na2SO4)
Mean Reproduction
29.8
31.2
31.2
30.8
27.0
31.8
30.6
31.4
Percent Reduction Percent Reduction
(compared to control) (compared to baseline)
-4.7
-4.7
-3.36
9.40
-6.71
-2.68
-5.37
Treatment(test initiated 4/22/04) Mean Reproduction Percent Reduction
sample collected 3/29/04 (compared to control)
Control
27.2
Treatment 1(100%ef
.. baseline)
29.0
-6.62
Treatment 2 (100%
eff. spiked)
21.6
20.59
Treatment 3 (100%
spiked, 0.5 mg/L EDTA)
26.8
1.47
Treatment 4 (100%
spiked, 3.0 mg/L EDTA)
25.6
5.88
Treatment 5 (100%
spiked, 8.0 mg/L EDTA)
21.2
22.05
Treatment 6 (100%
spiked, 0.5 mg/L Na2SO4)
23.6
13.24
Treatment 7 (100%
spiked, 3.0 mg/L Na2SO4)
25.6
5.88
Treatment 8 (100% spiked, 8.0 mg/L Na2SO4)
25.0
8.09
0.0
1.28
13.46
-1.92
1.92
-0.64
Percent Reduction
(compared to baseline)
25.52
7.59
11.72
26.90
18.62
11.72
13.79

Treatment (test initiated 6/23/04)
Mean Reproduction
Percent Reduction
sample collected 6/7/04
(compared to baseline)
Control
22.2
Treatment 1 (100% effluent, baseline)
24.4
-9.91 (compared to control)
Treatment 2 (0.5 mg/L EDTA)
24.4
0.0
Treatment 3 (3.0 mg/L EDTA)
25.6
4.92
Treatment 4 (8.0 mg/L EDTA)
24.4
0.0
Treatment 5 (0.5 mg/L Na2SO4)
26.6
-9.02
Treatment 6 (3.0 mg/L Na2SO4)
25.0
-2.46
Treatment 7 (8.0 mg/L Na2SO4)
24.0
-1.64
Treatment (test initiated 7/7/04)
Mean Reproduction
Percent Reduction
sample collected 6/7/04
(compared to baseline)
Control
20.8
Treatment 1 (100% eff. baseline)
22.8
-9.62 (compared to control)
Treatment 2 (100% eff. spiked)
14.2
37.72
Treatment 3 (100% spiked, 0.5 mg/L EDTA)
22.8
0.0
Treatment 4 (100% spiked, 3.0 mg/L EDTA)
20.6
9.65
Treatment 5 (100% spiked, 8.0 mg/L EDTA)
13.4
41.23
Treatment 6 (100% spiked, 0.5 mg/L Na2SO4)
21.2
7.02
Treatment 7 (100% spiked, 3.0 mg/L Na2SO4)
22.0
3.51
Treatment 8 (100% spiked, 8.0 mg/L Na2SO4)
23.8
-4.39
Treatment (test initiated 8/25/04)
Mean Reproduction
Percent Reduction
sample collected 8/23/04
(compared to baseline)
Control
20.2
Treatment 1 (100% effluent, baseline)
24.6
-21.8 (compared to control)
Treatment 2 (0.5 mg/L EDTA)
24.0
2.44
Treatment 3 (3.0 mg/L EDTA)
25.6
4.07
Treatment 4 (8.0 mg/L EDTA)
21.6
12.2
Treatment 5 (0.5 mg/L Na2SO4)
23.6
4.07
Treatment 6 (3.0 mg/L Na2SO4)
23.8
3.25
Treatment 7 (8.0 mg/L Na2SO4)
23.4
4.88
Treatment (test initiated 9/l/04)
Mean Reproduction
Percent Reduction
sample collected 8/23/04
(compared to baseline)
Control
21.8
Treatment 1 (100% eff. baseline)
23.4
-7.34 (compared to control)
Treatment 2 (100% eff. spiked)
15.2
35.04
Treatment 3 (100% spiked, 0.5 mg/L EDTA)
24.0
-2.56
Treatment 4 (100% spiked, 3.0 mg/L EDTA)
23.6
-0.86
Treatment 5 (100% spiked, 8.0 mg/L EDTA)
21.4
8.55
Treatment 6 (100% spiked, 0.5 mg/L Na2SO4)
19.6
16.2
Treatment 7 (100% spiked, 3.0 mg/L Na2SO4)
22.0
5.98
Treatment 8 (100% spiked, 8.0 mg/L Na2SO4)
16.0
31.62
The first round of TIE testing was conducted with the effluent sample collected on 3/29/04. Toxicity testing was
initiated on 3/31/04 and 4/22/04. Effluent sample copper and zinc concentrations measured 3.0 ug/L and 41.0 ug/L,
respectively. The investigators compared effluent treatments using the 3/29/04 effluent samples to the control. We

believe more useful information can be obtained by comparing the TIE treatment reproductions to the effluent baseline
reproduction. The far right column above titled "Percent Reduction (compared to baseline)" has been added to assist in
clarification. The report notes a significant reduction in reproduction with the 3/29/04 "spiked " sample and suggests the
reduction in reproduction (20.59%) is less than would be expected, possibly due to metals being complexed and
becoming less bio-available to the test organisms. The City notes a significant reduction in reproduction (22.05%) in the
8.0 mg/L EDTA "spiked' treatment and attributes the reduction to the presence of unbound EDTA. The reductions in
reproduction compared to the baseline are 25.52% and 26.90%, respectively.
The second round of TIE testing was conducted with the same effluent sample collected on 6/7/04. Toxicity
testing was initiated on 6/23/04 and 7/7/04. Effluent sample copper and zinc concentrations measured 2.0 ug/L and 33.0
ug/L, respectively. A slight increase in organism reproduction was noted in the 3.0 mg/L EDTA, 0.5 mg/L Sodium
Thiosulfate, 3.0 mg/L Sodium Thiosulfate and 8.0 mg/L Sodium Thiosulfate treatments. For "spiked" treatments, the
report notes equal reproduction in the 0.5 mg/L EDTA treatment and a slight increase in reproduction in the 8.0 mg/L
Sodium Thiosulfate treatment.
The third round of TIE testing was conducted with the same effluent sample collected on 8/23/04. Toxicity
testing was initiated on 8/25/04 and 9/l/04. Effluent sample copper and zinc concentrations measured 25.6 ug/L and 49.1
ug/L, respectively. A slight increase in organism reproduction was noted in the 3.0 mg/L EDTA treatment. For "spiked'
treatments, only the 0.5 mg/L and 3.0 mg/L EDTA treatments showed slight decrease in effluent toxicity.
All baseline(100% effluent) test results indicate an absence of toxicity. The City contends that effluent copper is
complexed and is not bio-available to the test organisms. The report concludes that neither this TIE nor historical data
supports the theory that copper and/or zinc are causative effluent toxicants.
The report attributes effluent toxicity during November and December 2003 to surfactants entering the
wastewater treatment plant; however, the report fails to mention what observations lead the investigators to this
conclusion, does not offer potential sources of surfactant contribution and does not suggest actions to be undertaken to
prevent refractory toxicity in the future. We point out that in addition to removing toxicity caused by metals, EDTA
reduces the acute toxicity of some cationic surfactants.
Our office supports innovative approaches to TIE work; however, after reviewing the TIE data, we question
what is gained by spiking a toxic effluent with copper and zinc and subsequent treatment manipulation.
A review of the facility's WET test results for NPDES compliance purposes shows no additional WET permit
limit violations since December 2003. TIE testing should be performed with toxic effluent samples. In situations where
an effluent is not toxic, we have allowed facilities under the Copper and Zinc Policy to submit a final report indicating
actions taken to address WET noncompliance and stating no additional WET permit limit violations occurred. In those
situations, facilities would remain subject to the policy should additional WET permit limit violations occur in the future.
In summary, in our opinion it can't be determined whether copper and/or zinc caused the facility's WET failures
in November and December of 2003. No WET failures occurred subsequently and no meaningful TIE work could be
conducted. The facility remains subject to the copper/zinc policy should WET failures occur in the future.
Please feel free to contact either Matt or me if 733-2136 if you have questions
cc: Steve Tedder — Winston-Salem Regional Office
Susan Wilson-NPDES Unit
Pretreatment Unit-DWQ
Mr. Morgan Huffman, WWTP Superintendent, 10 Salem Street, Thomasville, NC 27316-0638
Ms. Shannon Lund-Meritech, Inc., 642 Tamco Road, Reidsville, NC 27320
Kim Pierce, US EPA Region IV, Water Mgt. Div., 61 Forsyth St., SW, Atlanta,
Georgia 30303-8960
Aquatic Toxicology Unit Files
Central Files

CITY OF THOMASVILLE
North Carolina
UTILITIES DEPARTMENT
North Carolina Division of Water Quality
NOV 18 20(4
Environmental Sciences Branch Environmental Sciences F
Aquatic Toxicology Unit
1621 Mail Service Center
Raleigh, North Carolina 27699-1621
Subject: Copper and Zinc Action Level Policy Reply Option b) 3) Report
Dear Sirs, AJG��4 t l 2
hgv,nsoN CpuNr�
Please find enclosed three copies of the final report from the City of Thomasville
in response to the implementation of the Copper and Zinc Action Level Policy. We feel
that the testing performed and the enclosed report rule out copper and/or zinc as the
causes of the toxicity observed in November and December 2003.
I NOV 2 2 2004
i
DEAR -WATER QUALITY
POINT SOURCE BRANCH vs
Sincerely, /
'0 f/
Mo an Huffm '
WWTP Superintendent
City of Thomasville
City Hall 10 Salem Street
• P.O. Box 368
• Thomasville, N.C. 27361-0368
Telephone [phone] • Fax [phone]

tP e-Ub 611404-
Reply to Copper and Zinc Action Level Policy Implementation
Hamby Creek WWTP, Thomasville, NC
NCDENR has asserted that the toxicity observed in the City of Thomasville
Hamby Creek WWTP effluent in November and December 2003 was directly attributable
to the levels of copper and/or zinc in the effluent. The City of Thomasville Hamby Creek
WWTP has prepared this report to demonstrate the lack of toxicity directly attributable to
copper or zinc in the effluent. The results of the Toxicity Identification Evaluation phase
presented herein demonstrate that the assertion of copper/zinc toxicity is not only untrue,
but that the WWTP effluent may actually have a buffering affect against the potential
toxicity caused by these two metals. This is observed in the TIE test results that have
been spiked with the addition of copper and/or zinc. Historical data presented in
graphical form at the end of this document also demonstrate no correlation between
copper/zinc levels in the effluent and WET test failures. Observations by WWTP
personnel at the time of the toxic events in November and December 2003 indicate that
the toxicity was likely attributable to heavy inflows of surfactants to and through the
plant.
Following the implementation of the Copper and Zinc Action Level Policy by
NCDENR, the City of Thomasville WWTP contracted with Meritech, Inc. to run a series
of three toxicity test on the WWTP effluent using whole effluent, effluent samples spiked
with either EDTA or sodium thiosulfate at different levels, effluent samples spiked with
copper and zinc, and effluent samples spiked with copper and zinc and either EDTA or
sodium thiosulfate at different levels. All effluent samples were analyzed for copper and
zinc concentrations present before testing proceeded. This series of tests were run on
WWTP effluent samples collected on March 29, June 7, and August 23, 2004. Control
water used for the tests was filtered Lake Reidsville dilution water used by Meritech, Inc.
for toxicity testing.
In the first test effluent, collected on March 29, 2004, copper was found to be
present at a concentration of 3.Oug/L and zinc was present at a concentration of 41.0ug/L.
Both are below NOEC levels (12.0ug/L copper and 47.0ug/L zinc) as described in
"Quality Criteria for Water" (EPA Publication, 1986). On March 31, 2004 a toxicity test
series was started using control water, 100% effluent, and effluent treated with either
EDTA or sodium thiosutfate at concentrations of 0.5mg/L, 3.Omg/L, and 8.0mg/L.
Toxicity data, as presented in the attached original report from Meritech, Inc.,
demonstrates that there was no toxicity observed in the effluent. In point of fact, the
organisms in the 100% effluent sample reproduced better (4.7% reduction) than the
control.
A second test series was begun on April 22, 2004, using the same effluent sample
collected on March 29, 2004, using control water, 1000/6 effluent, effluent spiked with
20ug/L copper and 60ug/L zinc, and effluent spiked with 20ug/L copper and 60ug/L zinc
and treated with either EDTA or sodium thiosulfate at concentrations of 0.5mg/L, 3.0
mg/L, and 8.Omg/L. Again, no toxicity was observed in the 100% effluent sample and
the 100% effluent sample reproduced better (-6.62% reduction) than the control. In the
effluent sample spiked with 20ug/L copper and 60ug/L zinc there was a significant

reduction in reproduction of 20.59%. However, this is a less pronounced effect than
would be expected, indicating that these metals complexed with other elements present in
the effluent, making them less bio-available to the test organisms. A significant
reduction in reproduction of 22.05% was also observed in the effluent sample spiked with
20ug/L copper and 60ug/L zinc and 8.Omg/L EDTA. This is most likely attributable to
the presence of unbound EDTA.
In both test series run on the effluent sample collected on March 29, 2004 the
effluent demonstrated no toxicity and organisms in the whole effluent reproduced better
than in the control. The effluent sample spiked with 20ug/L copper and 60ug/L zinc
showed a significant reduction in reproduction of 20.59% compared to the control. No
toxicity was observed in the effluent sample collected on March 29, 2004 and the effluent
actually demonstrated a buffering effect against copper/zinc toxicity.
The second effluent sample for Toxicity Identification Evaluation copper and zinc
testing was collected on June 07, 2004. This effluent sample was tested for metals and
copper was found to be present at a concentration of 2.Oug/L and zinc at 33.Oug/L. Again,
both metals are below NOEC levels (12.0ug/L copper and 47.Oug/L zinc) as described in
"Quality Criteria for Water" (EPA Publication, 1986). On June 23, 2004 a toxicity test
series was started using control water, 100% effluent, and effluent treated with either
EDTA or sodium thiosulfate at concentrations of 0.5mg/L, 3.Omg/L, and 8.0mg/L.
Toxicity data, as presented in the attached original report from Meritech, Inc.,
demonstrates that again there was no toxicity observed in the effluent and again the
organisms in the 100% effluent sample reproduced better (-9.91% reduction) than the
control. The organisms in the effluent samples treated with 3.Omg/L EDTA, and
0.5mg/L, 3.Omg/L, and 8.Omg/L sodium thiosulfate all reproduced slightly better than the
100% effluent sample indicating a possible reduction by these treatments of some toxic
component present.
The second test series on the second effluent sample was begun on July 06, 2004
using the same effluent sample collected on June 07, 2004, using control water, 100%
effluent, effluent spiked with 20ug/L copper and 60ug/L zinc, and effluent spiked with
20ug/L copper and 60ug/L zinc and treated with either EDTA or sodium thiosulfate at
concentrations of 0.5mg/L, 3.0 mg/L, and 8.0mg/L. Again, no toxicity was observed in
the 100% effluent sample and the 100% effluent sample reproduced better (-9.62%
reduction) than the control. In the effluent sample spiked with 20ug/L copper and 60ug/1-
zinc there was a significant reduction in reproduction of 37.72% and 100% mortality on
day seven, as expected. The effluent sample spiked with 20ug/L copper and 60ug/L zinc
and treated with 0.5mg/L EDTA reproduced equal to the 100% effluent sample and the
effluent sample spiked with 20ug/L copper and 60ug/L zinc and treated with 8.0mg/L
sodium thiosulfate reproduced slightly better (4.39 reduction) than the 100% effluent
sample, indicating that these respective treatments removed the spiked copper/zinc
toxicity. All other samples spiked with 20ug/L copper and 60ug/L zinc and treated with
either 3.0 mg/L or 8.Omg/L EDTA or sodium thiosulfate at concentrations of 0.5mg/L or
3.0 mg/L exhibited reduced reproduction from the 100% effluent sample.
The third effluent sample for Toxicity Identification Evaluation copper and zinc
testing was collected on August 23, 2004. This effluent sample was tested for metals and
copper was found to be present at a concentration of 25.6ug/L and zinc at 49.1 ug/L.
Values for both metals are above NOEC levels (12.Oug/L copper and 47.Oug/L zinc) as
2

described in "Quality Criteria for Water" (EPA Publication, 1986). On August 25, 2004
a toxicity test series was started using control water, 100o/o effluent, and effluent treated
with either EDTA or sodium thiosulfate at concentrations of 0.5mg/L, 3.Omg/L, and
8.Omg/L. Toxicity data, as presented in the attached original report from Meritech, Inc.,
demonstrates that again there was no toxicity observed in the effluent and again the
organisms in the 1001/6 effluent sample reproduced better (-21.8% reduction) than the
control. The organisms in the effluent sample treated with 3.Omg/L EDTA reproduced
only slightly better (4.071/o reduction) than the organisms in the 100% effluent sample.
The organisms in the 100% effluent samples treated with 0.5mg/L and 8.Omg/L EDTA,
and 0.5mg/L, 3.Omg/L, and 8.Omg/L sodium thiosulfate all reproduced slightly worse
than the 100% effluent sample.
The second test series on the third effluent sample was begun on September 01,
2004 using the same effluent sample collected on August 23, 2004, using control water,
100% effluent, effluent spiked with 20ug/L copper and 60ug/L zinc, and effluent spiked
with 20ug/L copper and 60ug/L zinc and treated with either EDTA or sodium thiosulfate
at concentrations of 0.5mg/L, 3.0 mg/L, and 8.Omg/L. Again, no toxicity was observed
in the 100% effluent sample and the 100% effluent sample reproduced better (-7.34%
reduction) than the control. There was a significant reduction in reproduction in the
100% effluent sample spiked with 20ug/L copper and 60ug/L zinc of 35.04% indicating
the toxic effect these added metals concentrations had. In the 100% effluent samples
spiked with 20ug/L copper and 60ug/L zinc and treated with 0.5mg/L and 3.Omg/L
EDTA the organisms reproduced slightly better (-2.56% reduction and -0.86% reduction
respectively) than in the 100% effluent sample. In the 100% effluent samples spiked
with 20ug/L copper and 60ug/L zinc and treated with 8.Omg/L EDTA or 0.5mg/L
3.Omg/L, or 8.0mg/L sodium thiosulfate all organisms reproduced worse (8.55%, 16.2%,
5.98%, and 31.62% reduction respectively) than the 100% effluent sample.
In the case of all six test series run, the untreated 100% effluent sample
reproduced better than the control and exhibited no toxicity. Statistically significant
copper and zinc toxicity in the entire test series could only be observed in effluent
samples spiked with 20ug/L copper and 60ug/L zinc, with or with out the addition of
either EDTA or sodium thiosulfate. Even in the third effluent sample where copper was
found to be present at a concentration of 25.6ug/L and zinc at 49.1ug/L, both above the
NOEC levels, no toxicity was observed in the organisms in the 100% effluent sample. It
is apparent that even with levels above the NOEC values (12.Oug/L copper and 47.Oug/L
zinc) the copper and zinc present in the effluent are complexed with other elements
present in the effluent and therefore not bio-available to the organisms.
Copper levels in this run of three Toxicity Identification Evaluation test
series exceeded the prospective NPDES copper limit of 18ug/L on only one occasion
(25.6ug/L on August 23, 2004), but even this effluent sample exhibited no toxicity.
Effluent copper levels recorded closest to the toxic events observed in November and
December 2003 were 33ug/L and 2lug/L respectively, one being above and one below
the level shown here to have no toxic effect on organisms in the effluent. It is our
contention that the copper levels found in the effluent are complexed in a bound form and
not in a dissolved ionic form available to organisms in the effluent. Given the large
quantity of varying elements present in treatment plant process and effluent water, the
highly reactive nature of copper, and the difficulty in distinguishing between the forms of

copper present, it is highly unlikely and illogical to assume that copper present in the
effluent is in a toxic and highly reactive ionic form rather than bound in compounds and
complexes in harmless forms. Neither this TIE nor historical data (see attached graphs of
historical copper/zinc/WET/Phase II results) support the theory that copper has played a
role in toxicity events. Implementation of a copper limit on the Hamby Creek WWTP
NPDES permit would be a waste of resources.
Zinc levels in the three test series run and in the two months which triggered the
implementation of the Copper and Zinc Action Level Policy (November and December
2003) are well below the prospective NPDES zinc limit of 152ug/L suggested in the letter
of February 05, 2004 from Steve Tedder, Regional Water Quality Supervisor, Winston-
Salem. In fact, a review of DMR data dating back to February, 1999 finds no occasion
on which the effluent zinc level was as high as 152ug/L. There is no evidence, from
either this TIE study or historical data (see attached graphs of historical
copper/zinc/WET/Phase II results), to support the theory that zinc presents a toxicity
problem in the effluent from Hamby Creek WWTP. Implementation of zinc limit on
the Hamby Creek WWTP NPDES permit would he a waste of resources.
Again, observations by WWTP personnel at the time of the toxic events in
November and December 2003 indicate that the toxicity was likely attributable to heavy
inflows of surfactants to and through the plant. In conclusion, the City of Thomasville
feels that the above toxicity/metals study in conjunction with the historical data present
ample evidence that a) neither copper nor zinc in the Hamby Creek WWTP effluent are
to blame for the toxicity events observed in November and December 2003, and b)
implementation of copper or zinc limits on the Hamby Creek WWTP NPDES permit will
result only in expending resources that could be put to better use elsewhere.
0

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MER/TECH, INC.
ENVIRONMENTAL LABORATORIES
A Division of Water Technology and controls, Inc.
Thomasville W WTP is required by the state of North Carolina to demonstrate that
its effluent's toxicity is not caused by the presence of copper and zinc. Meritech, Inc.,
conducted this study on an effluent sample collected on March 29"' 2004. The sample
was first analyzed for both metals. Copper and zinc were present at concentrations of 3.0
µg/L and 41.0 µg/L respectively. Both metals were below NOEC values (i.e. 12.0 µg/L
for copper and 47.0 99/1 for zinc), as described in "Quality Criteria for Water" (EPA
Publication 1986). The metal analysis results are presented in attachment A. The
Thomasville effluent sample was also analyzed for hardness and suspended solids. The
sample's hardness was 76 mg/L of CaCO3 eq. and it contained 9.2 mg/L suspended
solids.
The first test series were started on March 31, 2004 using the aforementioned sample.
The test was set up with a control and seven treatments. 100% effluent was used in all
treatments. The test was set up as follows:
• Control: Filtered Lake Reidsville dilution water
• Treatment #1: 100% Effluent only
• Treatment #2: 100% Effluent + 0.5 mg/L EDTA
• Treatment #3: 100% Effluent + 3.0 mg/L EDTA
• Treatment #4: 100% Effluent + 8.0 mg/L EDTA
• Treatment #5: 100% Effluent + 0.5 mg/L Sodium Thiosulfate
• Treatment #6: 100% Effluent + 3.0 mg/L Sodium Thiosulfate
• Treatment #7: 100% Effluent + 8.0 mg/L Sodium Thiosulfate
Toxicity data is presented in attachment B. The test organisms in Treatment # 1
exhibited no toxicity and reproduced better than the control. In the sodium thiosulfate
treatments #5 thru #7 no toxic effect was observed and test organisms exposed in these
treatments produced better than the control. Similarly the EDTA treated treatments #2
and #3 also preformed better than control. A slight reduction (9.4%) in reproduction was
observed in Treatment #4, which was not significant.
Both treated and untreated 100% effluent concentrations did not show any toxicity.
A second test series was initiated on April 22, 2004 using the sample collected on
March 29, 2004. The test was set up with a control and eight treatments. 100% effluent
concentration was used in all treatment. The test was set up as follows:
642 Tamco Road • P.O- Box 27 Reidsville, NC 27320

MERI TECH, INC.
ENVIRONMENTAL LABORATORIES
A Division of Water Technology and Controls, Inc.
• Control: Filtered Lake Reidsville dilution water
• Treatment #1: 100% Effluent only
• Treatment #2: 100% Effluent + copper and zinc
• Treatment #3: 100% Effluent + copper and zinc + 0.5mg/L EDTA
• Treatment #4: 100% Effluent + copper and zinc + 3.Omg/L ETDA
• Treatment #5: 100% Effluent + copper and zinc + 8.0mg/L EDTA
• Treatment #6: 100% Effluent + copper and zinc + 0.5mg/L Sodium Thiosulfate
• Treatment #7: 100% Effluent + copper and zinc + 3.Omg/L Sodium Thiosulfate
• Treatment #8: 100% Effluent + copper and zinc + 8.Omg/L Sodium Thiosulfate
Toxicity data is presented in attachment C. Treatment #1 was set up with 100%
effluent only. No effluent toxicity was observed, and the reproduction was greater than
the control, which is similar to the first test. In Treatment #2, the sample was spiked with
20 µg/L copper and 60 µg/1 zinc. A significant reduction of 20.59% in reproduction was
observed. These metal concentrations should exhibit a pronounced toxic effect, but it is
possible that these metals complexed with suspended solids or TOC (total organic
carbon) making them less bio-available to test organisms. Treatments #3 thru #8 were
also spiked with copper and zinc. Then three different concentrations of EDTA and
Sodium Thiosulfate were used to chelate the metals. In EDTA treated treatments #3 and
#4, a slight reduction, 1.47% and 5.88% was observed respectively. In Treatment #5 a
significant reduction (22.05%) in reproduction was observed. This reduction may be
attributed to unbound EDTA. The Sodium Thiosulfate Treatments #6 thru #8 did not
show significant reduction in reproduction.
In both test series, 100% untreated sample, showed no toxicity and reproduced
better than the control. Therefore, these tests indicated that the effluent sample from
March 29, 2004 is non -toxic. Treatment #2 spiked with copper and zinc showed a
significant reduction (20.59%) in reproduction as compared to control. In Treatments #3
and #4 and #6 thru #8, both EDTA and Sodium Thiosulfate successfully removed or
reduced metal induced toxicity. Treatment #2 exhibited metal induced toxicity, as this
treatment was not treated with EDTA or Sodium Thiosulfate. No metal related toxicity
was observed in the effluent sample collected on March 29, 2004.
642 Tamco Road • P.O. Box 27 • Reidsville, NC 27320

AN �Cr 61 r-,,, ,I h
hoo•, �s(�r'dC�
Sample Name: Bio-Assay Diggested Acquired: 03/31/2004 16:17:23 Type: Unk
Method: IRIS REG 3 Mode: CONC Corr. Factor: 1.000000
User: admin Custom ID1: Custom ID2: Custom ID3:
Comment:
A13944
Sb2068
As1890
Ba4554
Be3131
B_2089
Cd2288
Cr2677
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
.1162
-.0057
.0093
.0082
-.0029
.2062
-.0007
.0036
.0022
.0022
.0034
.0001
.0005
.0022
.0001
.0019
1.880
38.27
36.73
1.705
18.35
1.068
10.34
52.25
.1168
-.0037
.0133
.0081
-.0023
.2051
-.0007
.0017
.1138
-.0081
.0074
.0082
-.0032
.2047
-.0007
.0036
.1181
-.0054
.0073
.0084
-.0033
.2087
-.0006
.0054
Co2286
Cu3247
Fe2599
Pb2203
Mn2576
Mo2020
Ni2316
Se1960
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
.0026
0.00/3 .0026
.1365
.0102
.1949
.0126
.0251
-.0060
.0002I
L .0005
.0047
.0050
.0013
.0007
.0006
.0029
8.516
19.33
3.445
49.38
.6780
5.781
2.461
47.80
.0023
.0030
.1341
.0119
.1940
.0125
.0246
0082
.0027
.0020
.1336
.0141
.1944
.0134
.0258
-.0027
.0027
.0026
.1420
.0045
.1965
.0120
.0248
-.0070
Ag3280
T11908
Sn1899
Ti3383
V_2924
Zn2138
Sc3353
ppm
ppm
ppm
ppm
ppm
ppm
ppm
.0001
.0247
-.0023
.0038
00100,M
.0410
229.1
.0004
.0035
.0038
.0004
.0007 1 q /4 .0003
1.8
297.1
14.33
167.0
11.96
75.10
.6118
.7786
0001
.0206
.0002
.0043
-.0016
.0413
227.0
-.0002
.0266
-.0066
.0035
-.0011
.0410
230.2
.0005
.0268
-.0004
.0035
-.0002
.0408
230.0
Sc2273
Sc3642
Cts/S
Cts/S
12.674
377.02
.048
2.08
.37512
.55147
12.621
378.19
12.688
378.24
12.713
374.62

Meritech
Crerpiodaphnia Chronic Reproduction Data Sheet
Client: �t f�i M s . Start Date: a-V-eq Time:
NPDES#: End Date: Al -7, pN Time:
Reviewed by:
Replicates
%Effluent Day# 1 2 3 4 5 6 7 8 9 in
Control
Tota
Tota
tota
Feo/ � 1u
f3•o+^�IL 7
Total
ry 3
ide f � iuv-d 5
ED9rA 7
Total
3
1"L ,�w�% 91 5
Total
14
31
Oro
R3rd Brood
Mean
Reoroduotion
31.4-
% RED. ij e
31•�
% RED. t{
% RED. 3 3 G
�i. n
%REDI q Ltv
%RED. —�'71

Meritech
y� Ceriodaphnia Chronic Reproduction Data Sheet
Client: fh;cYi�a M��� 4�� Start Date: Z �j.ar�i Time: /fg�pNj
NPDES#: �—
%Effluent Day# 1
Control
Tota
N. v
Tota
tota
c
7
Total
3
5
7
Total
3
5
7
Total
no Date: Y_-�_ o rP Time: , is- AAl
LI ReplicatReviewed by: 9
es
4 5 6 7 8 9 in %3d
s
i
r
S
Id
14
I
lG
it
i 3 2
2
3,�
Z
i sIl
6ro
9
s/«
s/l,
i 32D
31
3Z
L
r Brood
C.V. Mean
Reoroduclion
3b•t
% RED. LI--( $
31•f
% RED. S Sy-
% RED.B
% RED.
% RED.

a4t� L
Meritech
Ceriodaphnia
Chronic Reproduction
Data Sheet
Client:
NPDES#:
_ ,�
� �
�`�`�y�'"�
Start Date: - •?
2 • 0'+ Time:
IY 3o/rvl
End Date: y , 04
Time:
Reviewed by:
%Effluent Day#
1
2
3
4
Replicates
aF?Phi
3
5
67
8-9
10 %3rd Brood
Control 5
5
3
7
C.V. Mean
Total
9
6
025
Ree- —!production
a7. s.
3
iac g �Her�r 5
Total
3
r p
PDo% . S i 5
EAT 7
Total
_Dt,� t-�>7A"
Tot:
Ql �J-ATA
Totz
11
/6
4
iD
/5-
1
31
b'
l0
/o
l2
8
av
�3
2.2.
ss
ao
Ill `f if
0 i�7,
iS
Rg
.F-
% RED. 6•C2
% RED. MT67
U-$
% RED. 1.'f7
5- G
% R E D . 1 '6-27g
a1..2-
% RED.

Meritech
Cepriodaphnia Chronic Reproduction Data Sheet
Client:— �� Zn:L�S/' )� Start Date: _
NPDES#: �' End Date: L` ���Lr Time: !L•'�° �,�yyl
� .? 4j rf Time: _ q,.4 ° ¢M
Reviewed by: AF9QL
%Effluent Dav# 1 Z 3 Replicates
4 5 6 7 a
Control
Tota
�dil/. iku� 5
U Total
10or r��-.t'
cf to /
Tots
Tota
Tota
!r I 12 1 l3
3
5 ¢ 31r 4
7 i4 n r3 ILt !3
al 23 �> nL n� nn
%3rd erootl
C�
C.V. Mean
Reorotluction
a3.6
% RED. 13", -tf
% RED.=SL
�is•o
% RED.
% RED.[
% RED.[

Meritech, Inc.
Bioassay Sample Chain of Custody
642 Tamco Rd
Reidsville, NC 27320
[phone]
Client Information?
Client: sy(fe (•�byC:ee�G✓k1° PO#
Contact: 151 na NPDES #: 8 m
Address: l o Q rm ,'s d a . 3 Phone:
City: p AS Pipe Number:
County: a ✓. Uri State: Zip: 2,750
Type of Facility Generating Effluent: boll -Jo
Sample Information /
Sampling Site: {f &--11 604-tn -
Sample Type: Grab 1/ Composite # of Containers 2
Sampling Time: START Date: 2 -0 Time: 1000 pM
END Date: �3' 3 d- 0- 0/1 Time: I Q: d4l wpm
Sample Chilled (Covered with e�: Yes ✓ No
Collector's Name (Print): 4t4 Caosj&--
Collector's Signature: /Y I rX r
Toxicity Test Information
Test Required: Chronic: Test Organism: Ceriodaphnia Dubia:
Acute: Pimephales promelas:
IWC: Mysidopsis bahia:
1 oxiciry Test Concentration(s):
List Any Special Requirements: eaet r' - 7
Shipping Information
Relinquished By: OL
Date: 3- 0-0�
Time: 10,52.
AM PM
Received By: %,A,s,g,_
��—
Date:
Time: s
�PM
Relinquished By:
Date:
Time:
AMPM
Received By: Date: Time:..
Sample Temperature (C ): Method of Shipment: WTC(p/u) UPS
AM PM
Fed -Ex
Circle One Other:
Sample Receiving (Laboratory Use Only)
Received From: ` .
Received By: S• AD,c-t-,y Date: 3130JOU Time: /: 2L\AM M�
Samnle Temneratnre. (C • r1 � • t n • Co.. i. `. - - i

MERI TECH, INC,
ENVIRONMENTAL LABORATORIES
A Division of Water Technology and Controls, Inc.
Thomasville W WTP is required by the state of North Carolina to demonstrate that
its effluent's toxicity is not caused by the presence of copper and zinc. Meritech, Inc.,
conducted this study on an effluent sample collected on June 7, 2004. The sample was
first analyzed for both metals. Copper and zinc were present at concentrations of 2.0
µg/L and 33.0 µg/L respectively. Both metals were below NOEC values (i.e. 12.0 µg/L
for copper and 47.0 µg/1 for zinc), as described in "Quality Criteria for Water" (EPA
Publication 1986). The metal analysis results are presented in attachment A.
The first test series were started on June 23, 2004 using the aforementioned sample.
The test was setup with a control and seven treatments. 100% effluent was used in all
treatments. The test was set up as follows:
• Control: Filtered Lake Reidsville dilution water
• Treatment #1: 100% Effluent only
• Treatment #2: 100% Effluent + 0.5 mg/L EDTA
• Treatment #3:
100% Effluent +
3.0 mg/L EDTA
• Treatment #4:
100% Effluent +
8.0 mg/L EDTA
• Treatment #5:
100% Effluent +
0.5 mg/L Sodium Thiosulfate
• Treatment #6:
100% Effluent +
3.0 mg/L Sodium Thiosulfate
• Treatment #7:
100% Effluent +
8.0 mg/L Sodium Thiosulfate
Toxicity data is presented in attachment B. The test organisms in Treatment #1
exhibited no toxicity and reproduced better than the control. In treatments #2 thru #4
treated with EDTA, no toxic effect was observed and the test organisms produced better
or equal to treatment #1 (untreated 100% effluent) In the sodium thiosulfate treatments
#5 thru #7 no toxic effect was observed and test organisms exposed in these treatments
produced better than treatment #1. Both treated and untreated 100% effluent
concentrations did not show any toxicity.
A second test series was initiated on July 7, 2004 using the same sample collected
on June 7, 2004. The test was set up with a control and eight treatments. 100% effluent
concentration was used in all treatment. Treatments #2 thru #9 were spiked with 20 µg/L
copper and 60 µg/L zinc concentrations. The test was set up as follows:
642 Tamco Road • P.O. Box 27 • Reidsville, NC 27320

MER/TECH, INC.
ENVIRONMENTAL LABORATORIES
A Division of Water Technology and Controls, Inc.
• Control: Filtered Lake Reidsville dilution water
• Treatment # 1: 100% Effluent only
• Treatment #2: 100% Effluent + copper and zinc
• Treatment #3: 100% Effluent + copper and zinc + 0.5mg/L EDTA
• Treatment #4: 100% Effluent + copper and zinc + 3.0mg/L ETDA
• Treatment #5: 100% Effluent + copper and zinc + 8.0mg/L EDTA
• Treatment #6: 100% Effluent + copper and zinc + 0.5mg/L Sodium Thiosulfate
• Treatment #7: 100% Effluent + copper and zinc + 3.0mg/L Sodium Thiosulfate
• Treatment #8: 100% Effluent + copper and zinc + 8.0mg/L Sodium Thiosulfate
Toxicity data is presented in attachment C. Treatment #1 was set up with 100%
effluent _only in order to generate base -line data. No effluent toxicity was observed, and
the reproduction was greater than the control, which is similar to the first test. In
Treatment #2, the sample was spiked with 20 µg/L copper and 60 µg/L zinc. A
significant reduction of 37.72% in reproduction was observed. Treatment #2 also
exhibited 100% mortality on day seven, which is to be expected with these concentrations
of metals (copper and zinc). Treatments #3 thm #8 were also spiked with copper and
zinc. Then three different concentrations of EDTA and Sodium Thiosulfate were used to
chelate the metals. In EDTA treated treatment #3, with 0.5 mg/L EDTA no reduction in
reproduction was observed and reproduction was equal to that of treatment #1. In EDTA
treated Treatment #4, a slight reduction of 9.65% was observed. In Treatment #5 a
significant reduction (41.23%) in reproduction was observed. This reduction may be
attributed to unbound EDTA. The Sodium Thiosulfate Treatments #6 and #7 showed a
slight reduction of 7.02% and 3.51 % respectively. Treatment #8 exhibited no toxic effect
from the metals and reproduction was greater than Treatment #1.
In both test series, 100% untreated sample, showed no toxicity and reproduced
better than the control. Therefore, these tests indicated that the effluent sample from June
7, 2004 is non -toxic. Treatment #2 spiked with copper and zinc exhibited metal induced
toxicity. In this treatment there was a significant reduction of 37.72% in reproduction as
well as 100% mortality occurring on day seven. In Treatments #3 and #4 and #6 thru #8,
metals induced toxicity was removed or reduced by the addition of EDTA (treatments #3
and #4) and by Sodium Thiosulfate in Treatments #6 thru #8. Treatment #5 showed
toxicity but this can be attributed to unbound EDTA. Treatment #2 exhibited metal
induced toxicity, as this treatment was not treated with EDTA or Sodium Thiosulfate. No
metal related toxicity was observed in the effluent sample collected on June 7, 2004.
642 Tamco Road • P.O. Box 27 • Reidsville, NC 27320

,4#4 tr er, d,.)1
Sample Name:
Bio-Assay
Diggested
06/21/2004 15:32:59
Type: Unk
Method: IRIS REG
4 Mode: CONC
Corr. Factor: 1.000000
User: admin
Custom
ID1:
Custom ID2:
Custom ID3:
Comment:
=lem
A13944
Sb2068
As1890
Ba4554
Be3131
B_2089
Cd2288
Cr2677
Jnits
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
4vg
.1101
-.0100
.0078
.0028
.0005
.4287
-.0005
-.0040
3tddev
.0021
.0007
.0025
.0001
.0005
.0046
.0003
.0008
%oRSD
1.906
6.968
31.74
2.619
109.2
1.067
56.08
19.64
t1
.1116
-.0094
.0103
.0028
-.0001
.4238
-.0002
-.0034
12
.1110
-.0107
.0054
.0028
.0005
.4328
-.0004
-.0038
3
.1077
-.0097
.0076
.0027
.0009
.4295
-.0007
-.0049
Jem
Co2286
Cu3247
In2306
Fe2599
Pb2203
Mn2576
Mo2020
Ni2316
Inits
ppm
�'ooZ
ppm
ppm
ppm
ppm
ppm
ppm
ppm
vg
.0023
.0024
.0017
.0289
-.0162
-.0001
-.0017
-.0020
tddev
.0001
.0005
.0017
.0014
.0029
.0001
.0009
.0002
)RSD
4.552
21.23
100.7
4.802
18.09
140.1
54.87
8.749
1
.0023
.0018
.0009
.0304
-.0134
-.0001
-.0017
-.0020
?
.0022
.0025
.0006
.0287
-.0193
.0000
-.0026
-.0022
3
.0025
.0028
.0036
.0276
-.0160
-.0002
-.0007
-.0018
am
P_1774
P_1782
Se1960
Ag3280
T11908
Sn1899
Ti3383
V_2924
-)its
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ppm
ig
1.925
1.901
-.0059
-.0013
.0808
.0014
-.0009
.0036
ddev
.013
.018
.0009
.0003
.0031
.0009
.0009
.0009
RSD
.6863
.9742
15.85
24.98
3.859
66.69
102.1
25.72
1.911
1.883
-.0062
-.0012
.0826
.0023
-.0017
.0036
1.936
1.920
-.0048
-.0017
.0826
.0005
.0001
.0027
1.928
1.900
-.0066
-.0011
.0772
.0012
-.0011
.0046
>m
Zn2138
tits
ppm
g 0• a 3 3
.0330
idev
.0005
RSD
1.485
.0325
.0332
.0334

A77110INEN % " C "
Meritech
Cerppiodaphnia Chronic Reproduction Data Sheet
Client: ?f,y„5 -, 1_J. , r U r Start Date: o
NPDES#: o „ �' �' `f Time:
t�,RJJtF1� End Date: '%'•13 fit! Time:
f, i n/1m
Reviewed by:
°rrnuent uay#
4 5 6 7
Control
Tota
9 n 1C .1
Tota
Tota
• cv 14 7
Total
3
c ED-FA 7
Total
3
c D 7
Total
3
1
`oZ D
oRy
71
1 a�
I
Sn
a•
�D
L' �
9
al
OD
13
`f
��
JP
J
��
E D
/ 67
Cl
T 1/
/v
lC
ip
lv
a�
apt
az
311J'
a y-
Sf
2�
S
f;2
f o
&
%3rd Brood
C.V. Mean
Reproduction
e, 8
%RED. —9•%�, F4•r•-^C`"'�I"ar�
% RED. 39- 7A f2:
Al.9
% RED.
&, v
o, e 1
% RED. q• L5
VU k
13 � y- ��.� i'° f e >
% RED.

Meritech
Ceriodaphnia Chronic Reproduction Data Sheet
Client: sP_ (_ �F )?
NPDES#: yh,rOj, �i ` Start Date: �_ _ if Time:
n !2 if /�1r1
4pfe End Date: -it Time:
,Effluent Day# 1
'ontrol
Tota
Z' A
V
Tota
41,
,
Tota
Tota
R
7
Total
3
5
7
Total
2 3
. — I f : / e 6N1
Reviewed by: Ar
Replicates Uc �
4 5 6 7 a
3
5
7
I
3
-TO
y
To
20
z
L '
l�
i1
/-
/2
a
ir
al
22
°hard Brood
C.V. Mean
Reproduction
% RED.1i
;L- D 1
% RED. 3.5'1
2-3 1
`..µ'
% RED. + — 39
% RED.
% RED.

Pnncr.tino"L a
Meritech
Ceriodaphnia Chronic Reproduction Data Sheet
Client: 7� . c>r �� r 7 r�'rFp ��fI� Start Date:
NPDES#: '"7'�' LA3.044 Time: A-y.�,dtpt
End Date: •go•a7'f Time: g:3vflM
Reviewed by:
6Effluent Day# 1
Control
Tota
Tote
.7
,317Z
Tota
L i -I` 5
rH�/1 EIrT� 7
Total
3
5
Total
3
4�j-- 5
r1 k-T4"es' p
�JJ Total
Replicates
2 3 4 5 6 7 it
o .A
%Ord Brood
Mean
t, ( Reproduction
^�
��•�' C R
RED, G.Ci 1
% RED. O. mO '' !,
yp
% RED. FO 1°'G %
% RED.
%RED.

Meritech
Ceriodaphnia Chronic Reproduction Data Sheet
Client: .2;Z21v 11
NPDES#:
Start Date: f,-.a3,,7LF Time: End Date: ti 3 r h Time: c
Reviewed by:
)Effluent Day# 1
-ontrol
Totz
Tota
r
Tota
Tota
4
7
Tota
5
7
Total
Replicates
2 3 4 5 6 7 a
0
3 C
C:)
s 31�
L
sl u
C7
LD
'
Ifq
o
41/
!�
H
7� �
�7
iv
I'3
/3
/3
%3rd Brood
C.V. Mean
$'�ialo Reproduction
%RED.
% RED. — �• �� (�.� WW
% RED.B
% RED.B
% RED.

Title: Thomasville Metals Study 6/23/04
File: thomas62304 Transform: NO TRANSFORMATION
Kolmogorov Test for Normality
----------------------------------------------------------------------------
D = 0.1218 (p-value > 0.100)
D* = 0.7857
Critical D* = 1.035 (alpha = 0.01 , N = 40)
= 0.895 (alpha = 0.05 , N = 40)
Data PASS normality test (alpha = 0.01). Continue analysis.

Title: Thomasville Metals Study 6/23/04
File: thomas62304 Transform: NO TRANSFORMATION
Bartlett's Test for Homogeneity of Variance
----------------------------------------------------------------------------
Calculated Bl statistic = 4.5473 (p-value = 0.7150)
Data PASS B1 homogeneity test at 0.01 level. Continue analysis.
Critical B = 18.4753 (alpha = 0.01, df = 7)
= 14.0671 (alpha = 0.05, df =.7)

Title: Thomasville Metals Study 6/23/04
File: thomas62304 Transform: NO TRANSFORMATION
Summary Statistics on Data TABLE 1 of 2
----------------------------------------------------------------------------
GRP
IDENTIFICATION
N
MIN
MAX
MEAN
---
1
----------------
Control
----
5
----------
20.0000
----------
25.0000
----------
22.2000
2
100% Eff
5
23.0000
26.0000
24.4000
3
100%Eff+0.5EDTA
5
22.0000
25.0000
24.4000
4
100%Eff+3.OEDTA
5
23.0000
27.0000
25.6000
5
100%Eff+8.OEDTA
5
23.0000
26.0000
24.4000
6
100%Eff+0.5Thio
5
25.0000
28.0000
26.6000
7
100%Eff+3.OThic
5
24.0000
26.0000
25.0000
8
----------------------------------------------------------------------------
100%Eff+8.OThio
5
22.0000
26.0000
24.0000
Title: Thomasville Metals Study 6/23/04
File: thomas62304 Transform: NO TRANSFORMATION
Summary Statistics on Data TABLE 2 of 2
----------------------------------------------------------------------------
GRP
IDENTIFICATION
VARIANCE
SD
SEM
C.V. %
---
1
---------------- --------------
Control
3.7000
----------
1.9235
----------
0.8602
----------
8.6646
2
100% Eff
1.3000
1.1402
0.5099
4.6729
3
100%Eff+0.5EDTA
1.8000
1.3416
0.6000
5.4985
4
100%Eff+3.OEDTA
2.3000
1.5166
0.6782
5.9241
5
100%Eff+8.OEDTA
1.3000
1.1402
0.5099
4.6729
6
100%Eff+0.5Thio
1.8000
1.3416
0.6000
5.0438
7
100%Eff+3.OThio
0.5000
0.7071
0.3162
2.8284
8
----------------------------------------------------------------------------
100%Eff+8.OThio
3.5000
1.8708
0.8367
7.7951

Title: Thomasville Metals Study 6/23/04
File: thomas62304 Transform: NO TRANSFORMATION
ANOVA Table
----------------------------------------------------------------------------
SOURCE DF SS MS F
----------------------------------------------------------------------------
Between 7 56.9750 8.1393 4.0194
Within (Error) 32 64.8000 2.0250
----------------------------------------------------------------------------
Total 39 121.7750
----------------------------------------------------------------------------
(p-value = 0.0029)
Critical F = 3.2583 (alpha = 0.01, df = 7,32)
= 2.3127 (alpha = 0.05, df = 7,32)
Since F > Critical F REJECT Ho: All equal (alpha = 0.05)

Title:
Thomasville Metals
Study 6/23/04
File:
thomas62304
Transform:
NO
TRANSFORMATION
Dunnett's Test -
----------------------------------------------------------------------------
TABLE 1 OF 2
Ho:Control<Treatment
TRANSFORMED MEAN
CALCULATED
IN SIG
GROUP
-------------------------
IDENTIFICATION
MEAN ORIGINAL UNITS
T STAT 0.05
1
Control
-----------------------------
22.2000
22.2000
------ ---
2
100% Eff
24.4000
24.4000
-2.4444
3
100%Eff+0.5EDTA
24.4000
24.4000
-2.4444
4
100%Eff+3.OEDTA
25.6000
25.6000
-3.7778
5
100%Eff+8.0EDTA
24.4000
24.4000
-2.4444
6
100%Eff+0.5Thio
26.6000
26.6000
-4.8889
7
100%Eff+3.OThio
25.0000
25.0000
-3.1111
8
----------------------------------------------------------------------------
100%Eff+8.OThio
24.0000
24.0000
-2.0000
Dunnett
critical value = 2.4500
(1 Tailed, alpha
= 0.05, df
[used] = 7,30)
(Actual df = 7,32)
Title: Thomasville Metals Study 6/23/04
File:-thomas62304 Transform:
Dunnett's Test - TABLE 2 OF 2
NO TRANSFORMATION
Ho:Control<Treatment
----------------------------------------------------------------------------
NUM OF
MIN SIG DIFF
% OF
DIFFERENCE
GROUP
-------------------------
IDENTIFICATION
REPS
(IN ORIG. UNITS)
CONTROL
FROM CONTROL
1
Control
-------
5
----------------
-------
------------
2
100% Eff
5
2.2050
9.9
-2.2000
3
100%Eff+0.5EDTA
5
2.2050
9.9
-2.2000
4
100%Eff+3.OEDTA
5
2.2050
9.9
-3.4000
5
100%Eff+8.0EDTA
5
2.2050
9.9
-2.2000
6
100%Eff+0.5Thio
5
2.2050
9.9
-4.4000
7
100%Eff+3.OThio
5
2.2050
9.9
-2.8000
8
----------------------------------------------------------------------------
100%Eff+8.OThio
5
2.2050
9.9
-1.8000

Meritech, Inc. Obaeb
Bioassay Sample Chain of Custody DU
Tameo Rd
Reidsville, NC 27320
[phone]
Client Information
Client: Tkurm1 v; l e Qw)i P PO
Contact: 'a t r' NPDES #: C r7 112,-
Address: 0 (, 10 4I ir. Phone: 33E-k7S-�r141
City: ks Pipe Number: 06t
County: 1�+�,dsu,, State: L Zip: 273r,)
Type of Facility Generating Effluent
Sample Information
Sampling Site: ill L..,d,
Sample Type: Grab Composite # of Contain sss 7--
Sampling Time: START Date:—( -7-!t Time: 1 d p d PM
END Date: Time: (d PM
Sample Chilled (Covered with Ice): Yes No
Collector's Name (Print): A's� e;
Collector's Signature: 11t,,,124.
Toxicity Test Information
Test Required: Chronic: Test Organism: Ceriodaphnia Dubia: v"
Acute: Pimephales promelas:
(' to -TX T- - Mysidopsis bahia:
rwtr: d ��
i oxicity Test Concentration(s):
List Any Special Requirements:
Shipping Information
Relinquished By Date: is-0 Time:�M
Received By: I 6a Date: ,0 Time: /VJ, T M
Relinquished By: Date: Time: AM PM
Received By: Date: Time:: AM PM
Sample Temperature (C ): Method of Shipment: WTC(p/u) UPS Fed -Ex
Circle One Other: ('_our' ey-
Sample Receiving (Laboratory Use Only)
Received From:��L
Received By: 1 G ^ Date: CX f Time: AMQ)

M, RI TECH, INt„
ENVIRONMENTAL LABORATORIES
A Division of Water Technology and Controls, Inc.
Thomasville W WTP is required by the State of North Carolina to demonstrate
that its effluent's toxicity is not caused by the presence of copper and zinc. Meritech,
Inc., conducted the study on an effluent sample collected on 8/23/04. The sample was
analyzed for metals. Copper and zinc were present at concentrations of 25.6 µg/L and
49.1 µg/L, respectively. Both metals were above NOEC values (i.e., 12.0 µg/L for
copper and 47.0 µg(L for zinc), as described in "Quality Criteria for Water " (EPA
Publication 1986). The metals analysis results are presented in attachment A.
The first test series was start on 8/25/04, using the sample collected on 8/23/04.
An effluent concentration of 100% was used in all treatments. The test was set-up as
follows:
• Control (Lake Reidsville water)
• Treatment 1: 100% Effluent
• Treatment 2:
100% Effluent + 0.5 mg/L EDTA
• Treatment 3:
100% Effluent + 3.0 mg/L EDTA
• Treatment 4:
100% Effluent + 8.0 mg/L EDTA
• Treatment 5:
100% Effluent + 0.5 mg/L Thiosulfate
• Treatment 6:
100% Effluent + 3.0 mg/L Thiosulfate
• Treatment 7:
100% Effluent + 8.0 mg/L Thiosulfate
Toxicity data is presented in attachment B. The test organisms in Treatment 1
exhibited no toxicity and reproduced better than the control. In treatments 2 and 3,
treated with EDTA, a toxic effect was not observed, and the test organisms produced
better or equal to Treatment 1 (untreated 100% effluent). In Treatment 4, a 12.2%
reduction, which is not significant, was observed as compared to Treatment 1. In the
Sodium Thiosulfate treated samples, Treatments 5-7, toxicity was not observed, and a
slight, non -significant reduction in reproduction was observed as compared to Treatment
1. Both treated and untreated 100% effluent concentrations did not show any toxicity.
The second test series was initiated on 9/l/04, using the same effluent sample
collected on 8/23/04. An effluent concentration of 100% was used in all treatments.
Treatments 2-9 were spiked with 20 µg/L of copper and 60 µg/L of zinc. The test was
set-up as follows:
• Control (Lake Reidsville water)
• Treatment 1: 100% Effluent
• Treatment 2: 100% Effluent + copper + zinc
• Treatment 3: 1001/o Effluent + copper + zinc + 0.5 mg/L EDTA
642 Tamco Road • P.O. Box 27 • Reidsville, NC 27320

MERI TECH, INC.
ENVIRONMENTAL LABORATORIES
A Division of Water Technology and Controls, Inc.
• Treatment 4: 100% Effluent + copper + zinc + 3.0 mg/L EDTA
• Treatment 5: 100% Effluent + copper + zinc + 8.0 mg/L EDTA
• Treatment 6: 100% Effluent + copper + zinc + 0.5 mg/L Thiosulfate
• Treatment 7: 100% Effluent + copper + zinc + 3.0 mg/L Thiosulfate
• Treatment 8: 100% Effluent + copper + zinc + 8.0 mg/L Thiosulfate
Toxicity data is present in attachment C. Treatment 1 was set-up with 100%
effluent, to generate base -line data. Effluent toxicity was not observed in Treatment 1
and reproduction was greater than the control, which is similar to the first test series. In
Treatment 2, the sample was spiked with copper and zinc. A significant reduction of
35.0% in reproduction was observed as compared to Treatment 1, which can be attributed
to metal concentrations (copper and zinc). Treatments 3-8 were also spiked with copper
and zinc. Three different concentrations of EDTA and Sodium Thiosulfate were then
used to chelate the metals. In Treatments 3 and 4, which were treated 0.5 mg/L and 3.0
mg/L EDTA, respectively, no reduction in reproduction was observed, and the organisms
actually reproduced better than Treatment 1. In EDTA Treatment 5, a non -significant
reduction of 8.55% was observed.
Treatments 6-8 were treated with Sodium Thiosulfate. In Treatments 6 and 7 a
16.2% and 5.98% reduction in reproduction was observed, respectively. A significant
reduction of 31.6% was observed in Treatment 8. This reduction may be attributed to
unbound Sodium Thiosulfate.
In both test series, the 100% untreated effluent sample did not show any toxicity
and actually had a higher reproduction rate than the.control. Therefore, the effluent
sample of August 23, 2004 is considered to be non -toxic. Treatment 2, spiked with
copper and zinc, exhibited metal induced toxicity (i.e., a 35.0% reduction in
reproduction). In Treatments 3-5 and 6-7, metal induced toxicity was either removed or
reduced by the addition of EDTA (3-5) and Sodium Thiosulfate (6-7). Treatment 8
showed a significant reduction in reproduction, which can be attributed to unbound
Sodium Tbiosulfate.
642 Tamco Road • P.O. Box 27 • Reidsville, NC 27320

ample
Name: Bioassay Sample Acquired: 08/25/2004 14:55:04 Type: Unk
lethod:
IRIS REG 4 Mode: CONC
Corr. Factor:
1.000000
ser: admin Custom
ID1:
Custom ID2: Custom
ID3:
omment: Thmmasville
lem
A13944
Sb2068
As1890
Ba4554
Be3131
B_2089
Cd2288
Cr2677
nits
ppm Ppm ppm
0�0.00400.0i6
PPmG0,005
PPm
°.;23 ppm4o•0+Z
ppm
o •oltf ppm
vg
2037�1'o�S0022L0
.0156
-.0010
.3234
-.0005
.0136
tddev
.0088
.0005
.0024
.0002
.0000
.0193
.0002
.0014
)RSD
4.295
21.16
60.87
1.352
2.897
5.981
34.66
9.981
1
.2123
.0026
.0068
.0157
-.0010
.3430
-.0004
.0142
2
.2040
.0023
.0028
.0157
-.0010
.3228
-.0003
.0145
3
.1948
.0017
.0025
.0154
-.0010
.3043
-.0006
.0120
lem
CO2286
I� Cu3247
In2306
Fe2599
Pb2203
Mn2576
Mo2020
Ni2316
nits
Ppm
J PPm
ppm
0
PPm
0'191.1914
ppm
0'0100000
ppm
0-OW0644
ppm
0.023.02340
ppm
03�.0315
vg
G0 +9 •00120
026-0256
.0021
tddev
.0002
.0004
.0054
.0079
.0044
.0021
.0015
.0021
.RSD
17.38
1.666 253.1
4.101
28110.
3.307
6.242
6.610
25. I,
1
.0011
.0261
-.0040
.2003
.0035
.0667
.0251
.0338
.0014
.0254
.0044
.1885
.0015
.0641
.0228
.0311
3
.0253
.0061
.1855
-.0049
.0625
.0224
.0297
.0010
em
P_1774
Se1960
Ag3280
T11908
Sn1899
Ti3383
V_2924
Zn2138
PPM
Ppm G0 �-.0002
L0ppm
ppm00��.0491
02�
igs
a, 2.791 �'0.0l0.0002
0-o1�0220
057G090-.0002C0o�.0010
ddev
1 .061
.0039
.0003
.0011
.0029
.0007
.0009
0065
RSD
2.196
1798.
206.5
5.165
51.46
380.3
88.48
.9398
2.838
-.0028
.0002
.0224
-.0057
.0001
.0018
.0493
!
2.813
.0046
-.0005
.0207
-.0085
.0003
.0012
.0495
t
2.722
-.0011
-.0001
.0228
-.0027
-.0009
.0000
.0486
Sc2273 Sc3642
Cts/S
Cts/S
18.999
509.14
.149
7.87
.78498
1.5451
18.932
500.07
18.895
513.21
19.169
514.14

Meritech
Ceriodaphnia Chronic Reproduction Data Sheet
Client:�� ,�� t���Er�c Start Date:
NPDES#: ~( - ��c}C �`t Time: //
End Date: �i— t crt Time:
Reviewed by: � 7 L
An
Repticates—
%Cffluent Dav# 1 2 3 4 5 6 7 u o
%3rd Brood
DO
C.V. Mean
Reproduction
a.uc
Control
Tota
Tota
Tota
i
Tota
IMY LA't
i
Total
i
Total
% RED.
% RED.
�t'f •O
% RED. A° 4t}
Kb
%RED. I —f}•07
FT -
%RED. f°oG`�.r9

Meritech
Ceriodaphnia Chronic Reproduction Data Sheet
NPDES#:
ct Start Date: �j'-C;" Time:
End Date: q-i.flt: Time:
Reviewed by:
%Effluent Day# 1 Z 3 Replicates
4 5 6 7 a
Control
Tota
Tota
Tota
7
Total
5
7
Total
3
5
7
Total
I
G
a�-
�`l
13
10
%a
C-
WY
12—
I
io
L'
Lt
/q
/3
7
414
(%3rd Brood
I- I
C.V. Mean
Reorotluction
z3�t
% RED.
�3 g
% RED. ,3•,25-
3' Lj-
% RED. Z f QQ
% RED.[
% RED.

C
Meritech
` Ceriodaphnia Chronic Reproduction Data Sheet
Client: 0, _ Start Date:
NPDES#: (� — -17 Time: �pw/
End Date: G.-ott Time: gir F—
Reviewed by:
%Effluent Day# 1 2 3 4 5 Replicates
5 7 8 o an
Control
Tota
OZ P
Tota
7 {{
S�tKeo�
Tota
+,:� •�oSfhTA 7
pB //Tota
K'«J- 3.a EbTA 7
Total
3
mej S-a;=DTp 7
Total
%3rd Brood
tC.V. Mean
I J •'Q Reproduction
% RED.
I
% RED.
% RED.
% RED.
%RED. $•SS

Meritech
Ceriodaphnia Chronic Reproduction Data Sheet
Client: ThI,w_;" a St _c_ _ Start Date: C' I. py
NPDES#: l Time: a 91
yoEffluent Day# i
Control
Tota
}-0.57r-,o
Tota
Tota
cT 7
Total
3
5
7
Total
3
5
7
Total
End Date: Q.t-ay Time:
r---
Replicates Reviewed by:
l —
2 3 4 5 6 7 x o
i
I
gG
LIC
P
9
13
LHb
14
ILA
Lf 9
io
10
9
6
13
3
Lt ;7-
Ila
r
'Ord Brood
C�
C.V. Mean
Reoroduclion
Cr. tv
%RED. jw,a
%RED-1 S.Cip
IL c
% RED.1 31.(,X
% RED.
% RED.

Meritech, Inc. I
Bioassay Sample Chain of Custody
642 T
Reidsvillee,, NC NC 27 27320
(336) 3424748
Client InforP�atio
Client:as .//
Contact:
Address: () D parkPhone:
City: 4
PO # r�i(600
_ NPDES #: l02 ',!/-
E lr 75 -4 21ry
Pipe Number: OC) i
County: . o-s1 State:
WE Zip: oZ1a 1t
Type of Facility Generating Effluent:
Li
Sample Informations f L
Sampling Site: Lfi�aPrw�
Sample Type: Grab ✓ Composite # of Containers Z
Sampling Time: START Date: Time: 1 0 Dv PM
END Date: �SS"'�4 Time: 6mPM
Sample Chilled (Covered with e) Ye t. No
Collector's Name (Print): t:'
Collector's Signature: All
Toxicity Test Information
Test Required: Chronic:
% Acute:
M---,
IWC:
Toxicity Test Concentration(s):
List Any Special Requirements:
Test Organism: Ceriodaphnia Dubia: +/
Pimephales promelas:
Mysidopsis bahia:
►00 1
Shipping Information
Relinquished By: ✓T
Dater
Time:
AM PM
Received By:
Date:
Time:
AM PM
Relinquished By:
Date:
Time:
AM PM
Received By:
Date:
Time:
AM PM
Sample Temperature (C : Method of Shipment: WTC(p/u)
UPS Fed -Ex
Circle One
Other:
Sample Receiving (Laboratory Use Only)
Received From: A • �daz
/ Received By:a Date r% Time: la7>/
OAM PM
Sample Temperature (C�:1Q/ Samplg Condition 16

Title: Thomasville Zinc and Copper Study
File: Thom9104 Transform: NO TRANSFORMATION
Kolmogorov Test for Normality
-------------------------------------------------------------------
D = 0.0938 (p-value > 0.100)
D* = 0.6404
Critical D* = 1.035 (alpha = 0.01 , N = 45)
= 0.895 (alpha = 0.05 , N = 45)
-------------------------------------------------------------------
Data PASS normality test (alpha = 0.01). Continue analysis.

Title: Thomasville Zinc and Copper Study
File: Thom9104 Transform: NO TRANSFORMATION
Bartlett's Test for Homogeneity of Variance
---------------------------------------------------------------------
Calculated 31 statistic = 9.3383 (p-value = 0.3146)
Data PASS B1 homogeneity test at 0.01 level. Continue analysis.
-----------------------------------------------------------------------
Critical B = 20.0902 (alpha = 0.01, df = 8)
= 15.5073 (alpha = 0.05, df = 8)

Title: Thomasville Zinc and
Copper
Study
Transform:
NO TRANSFORMATION
File:
Thom9104
----------------------------------------------------------------------
Summary Statistics
on Data
TABLE
1 of 2
GRP
IDENTIFICATION
N
MIN
----------
MAX
----------
MEAN
---
1
----------------
Control
---- ----------
5
17.0000
26.0000
21.8000
2
100% Eff
5
21.0000
25.0000
23.4000
3
100% Eff Spiked
5
13.0000
19.0000
15.2000
4
Spiked+0.5EDTA
5
20.0000
26.0000
24.0000
5
Spiked+3.OEDTA
5
22.0000
25.0000
23.6000
6
Spiked+8.OEDTA
5
18.0000
27.0000
21.4000
7
Spiked+0.5Thio
5
14.0000
26.0000
19.6000
8
Spiked+3.OThio
5
20.0000
24.0000
22.0000
9
------------------------------------------------------------------------
Spiked+8.OThio
5
12.0000
21.0000
16.0000
Title: Thomasville Zinc and Copper Study
File: Thom9104 Transform: NO TRANSFORMATION
Summary Statistics on Data
TABLE 2 of
2
GRP
IDENTIFICATION
VARIANCE
SD
SEM
----------
C.V. %
----------
---
1
------------------------------
Control
10.7000
----------
3.2711
1.4629
15.0050
2
100% Eff
3.3000
1.8166
0.8124
7.7632
3
l00% Eff Spiked
5.7000
2.3875
1.0677
15.7070
4
Spiked+0.5EDTA
6.5000
2.5495
1.1402
10.6230
5
Spiked+3.OEDTA
1.8000
1.3416
0.6000
5.6849
6
Spiked+8.0EDTA
12.3000
3.5071
1.5684
16.3885
7
Spiked+0.5Thio
19.3000
4.3932
1.9647
22.4142
8
Spiked+3.OThio
2.5000
1.5811
0.7071
7.1870
9
--------------------------------------------------------------------
Spiked+8.OThio
16.5000
4.0620
1.8166
25.3876

Title: Thomasville Zinc and Copper Study
File: Thom9104 Transform: NO TRANSFORMATION
ANOVA Table
----------------------------------------------------------------------------
SOURCE
DF
SS
MS F
----------------------------------------------------------------------------
Between
8
417.3778
52.1722 5.9739
Within (Error)
36
314.4000
8.7333
----------------------------------------------------------------------------
Total
44
731.7778
--------------------------------------------------
(p-value = 0.0001)
Critical F =
3.0517 (alpha
= 0.01, df = 8,36)
=
2.2085 (alpha =
0.05, df = 8,36)
Since F > Critical F REJECT Ho: All equal (alpha = 0.05)

Title: Thomasville Zinc and Copper Study
File: Thom9104 Transform: NO TRANSFORMATION
Dunnett's Test -
TABLE 1 OF 2
Ho:Control<Treatment
----------------------------------------------------------------------------
TRANSFORMED
MEAN CALCULATED
IN SIG
GROUP
IDENTIFICATION
MEAN
ORIGINAL UNITS
------------------
T STAT 0.05
------ ---
-------------------------
1
Control
-----------
21.8000
21.8000
2
100% Eff
23.4000
23.4000
-0.8561
3
100% Eff Spiked
15.2000
15.2000
3.5312
4
Spiked+0.5EDTA
24.0000
24.0000
-1.1771
5
Spiked+3.OEDTA
23.6000
23.6000
-0.9631
6
Spiked+8.OEDTA
21.4000
21.4000
0.2140
7
Spiked+0.5Thio
19.6000
19.6000
1.1771
8
Spiked+3.OThio
22.0000
22.0000
-0.1070
9
Spiked+8.OThio
16.0000
16.0000
3_1032
-
---------------------------------------------------
Dunnett
critical value = 2.5000 (1 Tailed,
alpha = 0.05, df
[used] = 8,30)
(Actual
df = 8,36)
Title: Thomasville Zinc and Copper Study
File: Thom9104 Transform: NO TRANSFORMATION
Dunnett's Test -
TABLE 2
OF 2 Ho:Control<Treatment
----------------------------------------------------------------------------
NUM OF
MIN SIG DIFF
% OF
DIFFERENCE
GROUP
IDENTIFICATION
REPS
-------
(IN ORIG. UNITS)
----------------
CONTROL
-------
FROM CONTROL
------------
-----
1
--------------------
Control
5
2
100% Eff
5
4.6726
21.4
-1.6000
3
100% Eff Spiked
5
4.6726
21.4
6.6000
4
Spiked+0.5EDTA
5
4.6726
21.4
-2.2000
5
Spiked+3.OEDTA
5
4.6726
21.4
-1.8000
6
Spiked+8.OEDTA
5
4.6726
21.4
0.4000
7
Spiked+0.5Thio
5
4.6726
21.4
2.2000
8
Spiked+3.OThio
5
4.6726
21.4
-0.2000
9
----------------------------------------------------------------------------
Spiked+8.OThio
5
4.6726
21.4
5.8000