NC0005762_Report_19860311

DIVISION OF ENVIRONMENTAL MANAGEMENT
March 11, 1986
MEMORANDUM
TO:
George T. Everett
Dennis Ramsey
M.J. Noland
FROM: Steve W. Tedder
SUBJECT: Intensive Toxicological Evaluation: J.P. Stevens-Wagram
NPDES ¥fNC0005762, Scotland County
During the week of August 5-10, 1985 the Technical Services Branch
conducted an intensive toxicological evaluation of the subject facility.
The attached report details this investigation which included extensive
toxicological, biological, and chemical components.
If there are any questions, please contact myself or Ken Eagleson
at [phone] or Larry Ausley at [phone].
SWT:ps
cc w/attachment:
L.P. Benton, Jr.
Bob DeWeese
Ken Eagleson
Larry Ausley
Jim Overton
Meg Kerr
Jay Sauter
Ted Taylor

f1
J_P_ Stevens-Wagram
Toxicity Examination
NPDES#NC0005762
LNIIIIIIIIII"
IIIIIIUIIII
North Caroline Department of Natural
Resources & Community Development
I_ IIIINIII
NORTH CAROLINA DEPARTMENT OF NATURAL
RESOURCES AND COMMUNITY
DEVELOPMENT
WATER QUALITY SECTION
FEBRUARY, 1986

r
t
J.P. STEVENS - WAGRAM
TOXICITY EXAMINATION
NPDES A NO0005762
February 1986
NORTH CAROLINA DEPARTMENT OF NATURAL RESOURCES
AND COMMUNITY DEVELOPMENT
WATER QUALITY SECTION

TABLE OF CONTENTS
Page
List of Figures and Tables
Introduction 1
Toxicity Examination 2
Chemical Sampling 7
Benthic Macroinvertebrate Analyses 10
Conclusions 19
Recommendations 19
Appendix 20
Daphnia Dulex Test Procedure 21
96 Hour Flow -Through Test Procedure 22
Ceriodaphnia Reproduction Test Procedure 24
Oenthic Macroinvertebrate Procedure 25
tisrt of Definitions 26
-i-

LIST OF FIGURES
Page
Figure 1. Schematic Diagram of J.P. Stevens-Wagram Treatment Facilities3
Figure 2. Seven Day Ceriodaphnia Mean Cumulative Reproduction 5
Figure 3. Seven Day Ceriodaphnia Mortality 6
Figure 4. J.P. Stevens-Wagram Study Area and Sampling Stations 9
LIST OF TABLES
Page
Table 1. Sampling Site Descriptions, J.P. Stevens-Wagram Toxicity
Evaluation 10
Table 2. Chemical Results, J.P. Stevens-Wagram 11
Table 3_ Benthic Macroinvertebrate Sampling Station Descriptions 14
Table 4. Benthic Macroinvertebrate Taxa Richness Values 15
Table 5. Benthic Macroirt-vaertetrates Collected 16

INTRODUCTION
An on -site toxicity examination was conducted at the J.P. Stevens and Com-
pany-Wagram Plant (NC0005752) on August 5-10, 1985. This facility is located
approximately 3 miles from Wagram, N.C. in Scotland County.
This J.P. Stevens and Company facility consists of two plants, the Bob
Stevens Plant and the Scotland Plant. The Scotland Plant produces greige rolled
terry cloth used in the manufacture of towels_ This product is delivered to the
Bob Stevens Plant for bleaching, continuous dying, finishing, and screen print-
ing. Greige rolled carpet, received from the carpet plant in Aberdeen is also
included in the dying process.
This report details findings of chemical and biological sampling, including
the following:
1) 48 hr. static bioassay using Daphnia pulex on effluent samples to
determine acute toxicity.
2) 96 hr. flow -through acute bioassay using Pimephales promelas (fathead
minnows) performed on effluent collected at a holding pond prior to
the discharge pipe.
3) Chemical Samples collected of effluent, influent, and the recelving
stream.
4) Collection and analysis of macroinvertebrate samples to determine the
impact of the effluent on the receiving stream populations.
5) Seven-day Ceriodaphnia reproduction test to assess sub -lethal (chronic)
toxicity.
The facility's wastewater treatment plant receives process waste from the
Bob Stevens Plant and domestic wastes from both plants. The treatment plant
consists of screening, pH control by addition of sulfuric acid as necessary,
extended aeration with activated sludge, clarification, and a chlorine contact
chamber. Excess sludge is wasted to storage lagoons located on site. A land

application method of disposing of the sludge was being initiated at the time of
this investigation. A schematic of the facility appears in Figure 1.
Filter backwash and cooling water do not contribute to the waste stream
entering the treatment plant.
The J.P. Stevens-Wagram treatment plant discharges into the Lumber River
(Class "C-Swamp") in the Yadkin/Pee Dee River Basin. The 7010 (7 day, 10 year
low flow) for the Lumber River at this location is 101.5 cfs. The permitted flow
of the facility is 3.6 MGD. During 7010 low stream flow and average permitted
effluent flow conditions, the effluent from the J.P. Stevens-Wagram treatment
plant comprises 5.21% of the receiving stream. Results of past compliance samp-
ling inspections indicate compliance well within effluent limitations for all
parameters.
TOXICITY EXAMINATION
An on -site toxicity examination was performed at the J.P. Stevens and Com-
pany-Wagram Plant due to concerns arising from the presence of red coloration of
drinking water for the City of Lumberton, approximately 28 miles downstream of
the subject facility. Four 48 hour Daphnia pulex static tests were performed
prior to this examination on February 10, 1983, October 20, 1983, April 25, 1985,
and July 26, 1985. The resulting LC60's were 15%, 80%, 79% and NONE respec-
tively. The LC6* value is the effluent concentration lethal to 50% of the test
organisms over the indicated test period.
The flow -through bioassay was performed on effluent collected from the waste
stream leaving the holding pond. Although the facility has the capability for
chlorination, chlorine was not being added prior to or during this examination as
requested for the on -site testing. Dilution water for this test was obtained
from Juniper Creek at SR-1405 in Scotland County. Due to the low pH of the
dilution water, sodium bicarbonate was added to the dilution holding tank until
-2-

figure 1. Schematic Diagram of J.P. Stevens-Wagram Treatment Facilities
(2)
Sulfuric
Acid
Storage
Headworks
Bar Screen
pH Reduction
Secondary Clarifier
4
Influent
Station 02A
Extended
Aeration
Basin
Parshall flure
To L
River Station 02 Secondary Clarifier
-3- Bioassay Sampling Point

the pH was raised to near neutral (`6.8). To facilitate dissolving of the sodium
bicarbonate, a pump was used to continually circulate the water throughout the
tank. When a Toad of fresh dilution water was added to the tank, an appropriate
amount of sodium bicarbonate was also added.
The flow -through bioassay was initiated at 9:00 A.M. on August 6, 1985. The
test organisms, fathead minnows, were 61-66 days old and obtained from cultures
at the Aquatic Toxicology Laboratory. These fish were acclimated to Juniper
Creek water on August 2, 1985. The fathead minnows were transferred to test
chambers with dilution water approximately 17 hours prior to test initiation.
Due to an insufficient number of test fish, eight fish were placed in the control
(dilution water), 5%, 75%, and 100% effluent test chambers. Ten fish were placed
in the 10%, 25% and 50% effluent test chambers. Each concentration and the con-
trol had a replicate with the corresponding number of fish for each concentra-
tion. The toxicant delivery system cycled 488 times during the course of the
test. There was one mortality recorded in 100% effluent during the test. This
level of mortality (5%) is within that considered acceptable for the control
population and therefore is not considered significant_ The 96-hour LCso value
was determined to be greater than 100%. This value indicates there was no sig-
nificant acute toxicity to fathead minnows exhibited by the effluent during this
test.
A seven-day Ceriodaphnia static replacement bioassay was performed on dilu-
tions of effluent in order to assess sub -lethal toxicity. This test was initi-
ated at the Aquatic Toxicology Laboratory on October 25, 1985 and was concluded
October 31, 1985. Dilution water for this test was obtained from Lake Johnson
near Raleigh, N.C. This dilution water was chosen due to the low pH of the
dilution water used on -site (Juniper Creek). At the end of the seven-day test
period a significant reduction in reproductive success of the test organisms was
noted at the 75% effluent concentration. Reproduction similar to that of the
-4-

Figure 2. Seven Day Ceriodaphnia Mean Cumulative Reproduction
Mean Young
Produced
n
3
4
Mean Cumulative Reproduction
5
Day of Test
6
7
—5—

Figure 3. Seven Day Ceriodaphnia Mortality
100
T
0
X
C
A
N
T 10
V
0
L
u
M
E
LOG -CONCENTRATION VS MORTALITY
0 10 20 30 40 50 60
R MORTALITY
LC 50 =79%
70
80
90
100
-6-

controls was recorded tor all concentrations through 50%. Mean cumulative
reproduction is depicted in Figure 2. An LCso value of 78% was calculated based
on mortality results. The log-probit graph used to calculate this value is
depicted in Figure 3.
Since the on -site investigation was conducted, an additional self -monitoring
bioassay has been performed by this facility with a result of no significant
toxicity. This compares favorably with the absence of acute toxicity in a sample
analyzed by the Aquatic Toxicology Laboratory on October 28, 1985.
CHEMICAL SAMPLING
A series of chemical samples was collected during this evaluation and sent
to the Division of Environmental Management Chemistry Laboratory for analysis. A
description of the sampling stations is found in Table 1_ All samples were col-
lected as instantaneous grabs with the exception of Station 02 samples (effluent
bioassay sampling point) which were taken as 24 hour composites. Figure 4, a map
of the study area, illustrates sampling site locations. Results and summaries of
chemical analyzes are documented in Table 2.
Metals analyses of the J.P. Stevens effluent revealed elevated levels of
copper and zinc. Copper was reported at 80 ppb on August 8 and 200 ppb on August
10. Zinc was reported at 180 ppb on August 8 and 300 ppb on August 10. Nor-
mally, these levels would exhibit acute toxicity to both fathead minnows and
cladocerans (Daphnia pulex, and Ceriodaphnia dubia). Copper LCso's have been
reported as low as 75 ppb in 96 hour fathead minnow flow -through bioassays and
6.5 ppb in 48 hour Daphnia static bioassays.' Zinc LCso's have been reported as
low as 60 ppb in 96 hour fathead minnow bioassays and 100 ppb in 48 hour Daphnia
static bioassays_* Total residue was reported to be 1300 ppm on both sampling
' NTIS *P885-227023 Jan 85
a NTIS *PB81-117897 Oct 80
-7-

dates. These relatively high amounts of solids would make portions of the zinc
and copper biologically unavailable by way of adsorption to the solids.
There were no organic compounds identified in the J.P. Stevens effluent.
However, GC/MS analyses did detect 2 unidentified organic peaks at concentrations
of 35 and 120 ppb in the effluent sample collected on August 7-8 and 3 uniden-
tified organic peaks ranging from concentrations of 14 to 160 ppb in the August
9-10 sample.
Organic chemistry analyses of influent samples detected 9 unidentified peaks
in the August 8 sample ranging in concentration from 65 to 590 ppb and 14 uni-
dentified peaks in the August 10 sample ranging in concentration from 16 to 460
ppb. Tributyltin hydride (TBTH) was detected in the influent sample collected at
a level of 0.01 ppb. Although the presence of TBTH at any detectable level war-
rants some concern, it should be effectively treated by the J.P. Stevens wastew-
ater treatment plant if the amount detected is representative of normal levels
discharged to the plant. TBTH was not detected at any time in the effluent.
Hexamethyl Cyclotrisiloxane and Octamethyl Cyc!otetrasiloxane were detected in
the August 10 influent sample at levels of 8.5 and 14 ppb respectively. Uses or
toxicity levels have not been associated with these two compounds.
Upon comparing chemical parameters between sampling dates and within samp-
ling dates at the influent station (02A) and effluent station (02), it becomes
obvious that the J.P. Stevens effluent composition is of a highly variable
nature. For example, at Station 2 copper is reported at a level 0, 80 ppb on
August 8 and 200 ug/I on August 10. Both of these were 24 hour composite
samples. This sampling technique averages the high and low levels of chemicals
discharged over a 24 hour period. On August 8 the copper level from an instan-
taneous grab at the influent station (02A) was reported as 30 ppb. As noted
above, copper was found at 80 ppb in a composite sample taken from August 7-8 at
—8—

Figure 4. J.P. Stevens—Wagram Study Area and Sampling Sites
JP Stevens
Processing
Scale = 1 Rile

the effluent station (02)_ This illustrates the highly variable nature of the
J.P. Stevens effluent_
Table 1. Sampling Sites
Station 01 Lumber River at SR-1404 approximately 5600 m above the J.P. Ste-
vens-Wagram discharge. At this point the Lumber River is approxi-
mately 20 m wide with a gravel and sand substrate.
Station 02 J.P. Stevens effluent taken from the holding pond after clarifiers.
This is the bioassay sampling point for the Daphnia pulex static
tests, Ceriodaphnia reproduction tests, and the flow -through bioas-
say_
Station 02A J.P. Stevens influent collected just prior to bar screen.
Station 03 Lumber River at SR-1433 approximately 1600 m downstream of the J.P.
Stevens discharge. At this point the Lumber River is approximately
20 m wide with a predominantly sand substrate.
Station 04 Juniper Creek at SR-1405. Here the substrate is sandy with bottom
vegetation and approximately 4 m wide. This was the dilution water
location for all on -site bioassays.
BENTHIC MACROINVERTEBRATE ANALYSES
Benthic macroinvertebrate samples were collected on October 22, 1985. Sta-
tion 01 was located at Hoke County SR-1404, approximately 4 miles upstream of the
J_P. Stevens-Wagram discharge. Station 03 was located at Hoke County SR-1433,
approximately 1 mile downstream of the discharge. The sites were similar in
physical characteristics except for substrate: Station 01 had many gravel
riffles whereas Station 03 had very few (Table 3). These stations correspond to
chemical sampling sites 01 and 03.
—10—

Table 2. Chemistry Results, J.P. Stevens-Wagram
PERMITTED FLOU (MG0)
3.6
AVERAGE DISCOARGE (MGO)
2.12
7010 (CFS)
101.5
Chemical/Physical
Units
Utter Quality
STATION 01
STA1ION 02
STATION 02A
STATION 03
S1A1I0N 04
Analyses
Standards
15/08/08
15/08/08
85/08/08
15/18/18
85/08/01
800
PPM
1
1.2
>74
1.1
1.1
COO
PPM
20
210
1300
37
24
ColiforM: OF Fecal
/100 ml
<10
200
11000
20
<10
Residue T01A1
PPM
66
1300
1100
17
71
volatile
PPM
44
280
730
67
57
fixed
PPM
22
1000
350
20
14
Residue SUSPENDED
PPM
6
32
73
4
2
volatile
PPM
2
30
62
1
1
fixed
PPM
4
2
12
3
1
ph (standard units)
6.01.0
4.8
8.2
9.2
5.6
7.1
Acidity
PPM
15
3
16
5
Alkalinity
PPM
1
570
100
5
12
Grease and Oils
PPM
11/R
8
62
N/A
MIA
hardness
PPM
7
14
14
6
8
Phenols
FPO
<5
6
8
<5
<5
Sulfides
PPM
us
<0.1
I/O
M/A
N/A
Specific Caaductance
uhhos
25
17110
500
42
37
NM3
PPM
.15
.07
1
.03
<.01
TKO
PPM
.3
3.4
12
.4
.2
1102,1103
PPM
.12
.06
.52
.13
.1
P. total
PPM
r
.D6
3.5
5.2
.09
.02
Aluminum
PPI
408
200
600
30D
200
Cadmium
PP8
2 1
<10 1
<28
<20
<20
<11
Chromium
PPI
50
<5D
<50
<50
<50
<50
Copper
PPI
15(Al)1
<18
80
30
<20
<20
Iron
PPI
1000
910
280
300
800
400
Mercury
Pm
.2
<8.2
<0.2
<0.2
<0.2
<0.2
Nickel
III
50
<180
(100
<100
<100
<100
Lead
PPO
25
<100
<100
<100
<100
<100
Zinc
PPO
50(AL)
<211
180
50
<20
<20
18TM
PPI
.008
<.01
<.01
.01
<.02
<.02
Mexamethyl Cyclotrisiloxtne
PPS
NO
N0
N0
N0
NO
Octemethyl Cyclatetrasiloxane
PP8
NO
NO
NO
N0
N0
Petroleum 0i1
PPS
NB
98
1200
N0
NO
Vnidentlfied Peaks
!
1
2
9
3
N/A = Not analyzed
t = Values represent action levels as specified in .0211(b)(4) Fresh Water
Classification Standards

Table 2. Chemical Results, J.P. Stevens-Wagram (Cont.)
PERMITTED FLOU (Mfg)
3.6
AVERAGE OISOAROf (MOD)
N i 10 (c s)
101.5
Chemicnl/Physical
Units
STATION 01
STATION 02
STATION 02A
STATION 03
STATION 04
Predicted strew"
Analyses
15/00/10
15/11/10
15/10/10
85/01/10
15/08/10
conc. at 7010
100
PPM
Z.1
6.4
440
2.!
2.5
COO
PPM
41
160
1200
43
25
9.6315
Coliform: Of Fecal
/100 ml
N/A
M/A
N/A
N/A
N/A
Residue TOTAL
PPM
100
1300
940
15
76
Volatile
PPM
77
150
680
54
58
fixed
PPM
24
1100
260
31
18
Residue SUSPENDED
PPM
21
17
57
4
1
Volatile
PPM
3
15
57
1
1
fixed
PPM
11
2
<1
3
<1
pN (standard units)
4.7
1.1
7.1
5.4
6.5
Aridity
PPM
17
3
5
15
4
Alkalinity
PPM
<1
470
75
5
11
Grease and 011s
PPM
N/A
1
65
M/A
N/A
Madness
PPM
7
16
40
6
8
Phenols
PPO
<5
<5
<5
<5
<5
.313
Sulfides
PPM
N/A
<0.1
M/A
N/A
NIA
<.105
Specific Conductance
uIhos
25
1000
410
42
38
103
PPM
<0.01
.0!
.65
.03
<0.01
.004
TEN
PPM
.4
3
9.4
.5
.4
.117
102,1103
PPM
.08
.04
.3
.11
1.1
.813
P. total
PPM
.05
3.3
5
_09
.02
.177
81Mmiaum
PPB
500
400
500
200
200
15.630
CadNium
PPB
<20
<20
<20
<20
<20
<1.142
Chromium
PPB
<50
<50
220
<50
<50
<2.615
Copper
PPS
<20
ZOO
30
<20
<20
7.214
Iron
PPB
1000
200
700
1000
400
111.420
Mercury
PPS
<0.2
<0.2
<0.2
<0.2
<0.2
<_110
llCkel
PP1
<100
<100
<100
<100
<100
<5.210
lead
PPB
<100
<100
<100
<100
<100
<5.210
Zinc
PPO
30
300
50
30
30
12.514
T1TN
PPB
<.01
<.11
(.03
<_1Z
(.02
<.111
Mexamethyl Cyclotrisiloxene
PPB
M0
NO
8.5
ND
N0
Octamethyl Cyclatetrasilexane
PPB
NO
MO
14
NO
M0
Petroleum 0i1
PPB
NO
N0
NO
MO
N0
Imidentified Ptak
+
0
3
14
1
0
** = Values represent predicted instream concentrations using average effluent
concentrations and assuming upstream concentrations of 0.
-12-

Benthic macroinvertebrates were sampled using a standardized qualitative
collection technique (DEM 1983). The primary output from this collection tech-
nique is a tabulation of taxa richness, i.e., the number of different kinds of
animals present. Unstressed streams and rivers always have high taxa richness.
Various types of pollution will reduce or eliminate the more intolerant species,
producing lower taxa richness values. In-house criteria have been developed to
relate taxa richness to five water quality ratings or bioclassifications:
Excellent, Good, Good -Fair, Fair and Poor. Taxa richness values are calculated
both for all species (ST) and for the more intolerant groups (Ephemeroptera,
Plecoptera, Trichoptera: SEAT). The distribution and abundance of various
pollution "indicator" species also can be utilized to deduce changes in water
quality. Taxa richness values are summarized in Table 4. All taxa collected in
the survey are listed in Table 5.
Both stations were classified as Excellent based on benthological data.
Taxa richness values (STSepT) for Stations 01 and 03 were 89/34 and 90/29,
respectively. There was a slight decline in Trichoptera species below the
discharge, but this may reflect a loss of the gravel -riffle habitat at the latter
site rather than any toxic impact Many of the Trichopterans which were absent at
Station 03 were stone -cased species (Heliopsyche borealis, Goera sp_, and
Psilotreta sp.) that are associated with gravel substrate.
-13-

Table 3. Site Descriptions - Macroinvertebrate Analysis Sites.
Station * 01 03
Location SR-1404 SR-1433
County Hoke Hoke
Date 22-Oct-85 22-Oct-85
Depth (ml
Avg. 1.3 1.8
Max. 2+ 2+
Width (ml 15 22
Substrate (%l
Rubble/Bedrock - -
Gravel 30 10
Sand 60 80
Silt 10 10
Aufwuchs Low Mod
Canopy 30 30
Bank Erosion None None
-14-

Table 4 Taxa Richness by Group.
Station • 01 03
Group
*Ephemeroptera 6 8
*Plecoptera 10 7
*Trichoptera 18 14
Coleoptera 3 8
Odonata 8 10
Megaloptera 3 2
Diptera: Misc. 6 5
Diptera: Chiron. 21 20
Oligochaeta 5 4
Crustacea 3 3
Mollusca 1 2
Other 5 7
*Subtotal (EPT) 34 29
Total 89 90
*Unique Species' 13 5
Rating Excellent Excellent
' Found in only 1 out of 9 collections

Table 5 Total Taxa Collected
Station Number 01 03
EPHEMEROPTERA
Stenonema modestum A* A*
S. terminatum - C
Stenacron interpunctatum C C
Baetis pygmaeus - A
6. ephippiatus - R
Pseudocloeon sp. R -
Paraleptophlebia sp. A A
Siphloplectron sp. R -
Eurylophella bicolor C A
Hexagenia sp_ - R
PLECOPTERA
Acroneuria abnormis C A
A. carolinensis C -
Paragnetina fumosa C C
Pteronarcys dorsata C A
Perlinella sp. C R
Neoperla clymene R A
Helopicus bogaloosa C R
Isoperla sp. 10 C I. holochlora - R
Allocapnia R
Taeniopteryx sp. R
TRICHOPTERA
Cheumatopsyche sp. A C
Hydropsyche decalda A C
Macronema carolina - C
Chimarra sp. A* A*
8rachycentrus sp. A* A*
Oecetis sp. 1 C C
Nectopsyche exquisita R A
Triaenodes tarda C C
T. abus R R
Ceraclea transversa R
Helicopsyche borealis R -
Goera sp. R -
Psilotreta sp. R -
Agarodes libalis A R
Nyctiophylax nephophilus R R
Lype diversa C R
Neureclipsis sp. - R
Lepidostoma sp. R R
Anisocentropus pyraloides R -
Pychopsyche spp. R -

Station Number
01 03
COLEOPTERA
Macronychus glabratus C A
Ancyronyx variegate R R
Stenelmis sp. A A
Enochrus sp. - R
Deronectes griseostriatus - C
Laccophilus sp. - R
Peltodytes sp. - R
Gyrinus sp. - A
ODONATA
Macromia sp. C C
Boyeria vinosa A C
Basiaeschna janta R R
Nasiaeschra pentacantha - R
Neurocordulia virginiensis C R
Gomphus spp. C A
Progomphus obscurus - R
Argia spp. C A
Enallagma sp. A A
Calopteryx sp. C C
MEGALOPTERA
Sialis sp. R R
Corydalus cornutus C C
Nigronia serricornis C -
DIPTERA: MISC.
Simulium vittatum gr. C A
S. (Phosterodoros) sp. A -
Empididae R R
Hexatoma R -
Tipula sp. C C
Polymeda/ormosia - R
Palpomyia (complex) C C
DIPTERA: CHIRON.
Polypedilum halterale - R
P. scalaenum R -
P. illinoense C -
P. convictum - C
P. fallax R -
Phaenopsectra flavipes R R
P. sp. 2 R C
P. sp. 3 - R
Tribelos sp. - R
Stenochironomus sp. C C
Microtendipes sp_ 2 R R
Robackia claviger A C
Cryptochiromus fulvus - R
Demicryptochironomus sp. R -
Stelechomyia perpulchra R -
Rheotanytarsus sp. A A
-17-

Station Number 01 03
DIPTERA: CHIRON. (cont.)
Tanytarsus sp. C -
Cladotanytarsus sp. 2A - R
Ablabesmyia parajanta R R
A. ornata C C
Clinctanypus pinguis R -
Conchapelopia gr. R R
Labrundinia pilosella R -
Thienemaniellesp. C R
Corynoneura sp_ R R
Rheocricotopus sp. 1 A A
Eukiefferiella sp_ 3 C R
Cricotopus/Orthocladius sp. 8 - C
OLIGOCHAETA
Limnodrilus sp. R C
Huber speciosus R
Haplotaxis gordiodes R
Nais sp. R R
Stylaria lacustris - A
Lumbriculidae A A
CRUSTACEA
Palaemonetes paludosus R R
Crangonyx sp. R
Hyallela azteca R R
Procambarus sp. - R
MOLLUSCA
Pisidium sp_ R
Laevapex fuscum - A
Physella sp. 0 R
OTHER
Hydracarina R -
Sigara sp. C C
Ranatra sp. - R
Dugesia tigrina R -
Helobdella stagnales C C
H. elongate (?) R R
Batracobdetla phalera - R
Mooreobdella tetragon - R
Climacia areolis - R

CONCLUSIONS
Results of on -site toxicological evaluations conducted at J.P. Stevens-
Wagram's treatment facility do not predict an acutely toxic or chronically toxic
environment caused by J.P. Stevens' discharge in the Lumber River. Acute (168 hr
LC60 = 79%) and chronic (NOEC - 50%) toxicity observed in the Ceriodaphnia life
cycle test occurred at concentrations of effluent well above J.P. Stevens' low
flow instream waste concentration of 5.21%.
The metals copper and zinc were present in the effluent at levels which
normally would be expected to cause acute toxicity. However, this toxicity was
not exhibited, possibly due to adsorption of the metals to solids. Also, the
highly variable nature of the effluent may have prevented the organisms from
being exposed to the metals for a period of time sufficient to cause acute tox-
icity.
RECOMMENDATIONS
1) J.P. Stevens-Wagram should continue to perform self -monitoring acute toxicity
tests until the >90% LC60 target level has been achieved for three consecu-
tive months, at which time the facility should be required to perform the
Pass/Fail Ceriodaphnia reproduction bioassay. This test should be performed
monthly until such time as the effluent passes the test for three consecutive
months. If the test is failed two consecutive months, full range CeriodaDh-
nia tests should be accomplished to determine No Effect Levels on a quarterly
basis with Pass/Fail Ceriodaphnia tests continuing monthly until such time as
the Pass/Fail test is passed for three consecutive months.
2) Copper and zinc levels may at times cause exceedances of Water Quality Action
Levels in the Lumber River. These metals should be included as monitoring
requirements in the next J.P. Stevens' NPDES permit issuance.
-19-

APPENDIX
-20-

48 Hour Daphnia pulex Screening Bioassay
Aquatic Toxicology Group
N.C. Division of Environmental Management
The Aquatic Toxicology Group performs 48 hour static bioassays using the
cladoceran Daphnia 2u1ox to estimate the toxicity of waste discharge to aquatic
life in receiving streams. All test and sampling glassware and equipment are
washed with soap and hot water, then rinsed in nitric acid, acetone, and
distilled/deionized water, to remove all toxins end contaminants. Effluent
samples are collected by DEM Regional Office or Aquatic Toxicology personnel.
All samples are collected chilled and above chlorination unless otherwise
specified. Each sample is collected es a grab or 24 hour composite using an
automatic sampler and is sent chilled to the Aquatic Toxicology Laboratory by
State Courier or bus. The sample must be received with 72 hours after
collection.
The samples are prepared for testing by being thoroughly mixed, adjusted to
standard test temperature, and aerated if dissolved oxygen is below 40%
saturation. Chlorine is removed with sodium thiosulfate if applicable. The
effluent is then diluted with D. pulex culture water, typically to seven
concentrations (with replicates) from 0 to 90% effluent and initial pH and D0 are
recorded. Each test chamber receives 100 mis total volume and ten D. pulex test
organisms, 0-24 hours old. The test is conducted in a 20 degree centigrade
incubator with a 14:10 hour tight:dark cycle. Mortality of the D. pulex is
recorded after 48 hours, along with final pH, dissolved oxygen, and temperature.
A 48 hour LC60, or concentration of effluent lethal to 50% of the test
organisms in 48 hours, is calculated from the mortality data. An instream waste
concentration (IWC) for the effluent in the receiving stream is calculated using
the treatment system permitted flow and receiving stream 7010 flow. The LC6o and
IWC are then used to predict instream toxicity. 11 the effluent toxicity and/or
the IWC are high, a persistence bioassay may be conducted. This involves a
second 48 hour static 0. pulex bioassay on the same effluent sample after it has
been exposed to light and aeration for an additional 48 hours. If there is a
100% reduction in the LC60, the effluent is considered to be non -persistent.
Guidance Document: 1985. U.S.E.P.A Methods for Measuring the Acute Toxicity of
Effluents to Freshwater and Marine Organisms. Third edition (EPA/600/4-85/-13)
-21-

96 Hour On -Site Flow -Through Bioassay
Aquatic Toxicology Group
N.C. Division of Environmental Management
Candidacy for en on -site toxicity evaluation by the Aquatic Toxicology Group
is determined on the basis of acute toxicity of the effluent in comparison with
instream waste concentration. Acute toxicity is determined by a 48 hour
screening static bioassay.
For each on -site flow -through bioassay a pre -test inspection is performed in
order to:
1_) Determine appropriate areas for physical placement of the mobile
laboratory.
2.) Acquire proper equipment and installation needed for electrical
service.
3_) Determine appropriate areas for effluent sampling end equipment needed
for such. Sampling is done above chlorination unless otherwise
specified.
4.) Determine possible areas for dilution sampling (actual receiving waters
or other unstressed streams in the area) and equipment needed for such.
5.) Collect additional samples of effluent and possible dilution waters for
further static Daphnia pulex, acute and static renewal Ceriodaphnia
dubia_reproduction bioassays to determine the range of concentrations
of effluent for the flow -through bioassay, to test for potential
toxicity of possible dilution waters, and for fish acclimation to the
chosen dilution water.
6.) Determine route suitability to the facility for the mobile laboratory
(eg. low clearances, poor road conditions).
7_) Discuss test procedures and requirements with appropriate facility
personnel_
8.) Determine appropriate sampling sites and techniques for benthic
mecroinvertebrate surveys.
Upon actual arrival on -site with the mobile laboratory, dilution water is
obtained and dilution and effluent pumping systems are set up and tested. Six to
eight week-old fathead minnows are wet transferred to the test chambers
(containing approximately one liter of dilution water), ten fish to a chamber_
This transfer is accomplished five fish at a time in a randomized order to each
of the fourteen test chambers until two randomized sets of five have been
transferred to each chamber. Seven concentrations (with replicates) including a
control are used. The second day on -site the dilutor and the dilution and
effluent pumping systems are turned on and the fathead minnow flow -through
bioassay is begun. A water bath is utilized to bring the effluent and dilution
water to a constant 20 degrees centigrade. Test organisms are fed newly hatched
brine shrimp twice daily throughout the test.
A Ceriodaphnia static renewal reproduction bioassay using newborn organisms
is begun the first day on -site. The organisms are transferred to fresh dilution
and effluent solutions daily and initial and final pH and dissolved oxygen are
recorded. The number of young born per organism per day is recorded and mean
cumulative reproduction is calculated for each concentration. The rest ts
conducted in a 25 degree centigrade incubator with 16 light:8 dark photoperiod.
-22-

Test organisms are fed 0 1 ml of a fermented trout chow mixture per organism per
day.
Individual chemical/physical parameter meters are calibrated daily according
to DEM standards. Hydrolab systems measure and record dissolved oxygen. pH,
temperature, and specific conductance in the test chambers with the highest and
lowest concentration of effluent et 15 minute intervals throughout the test.
These systems are calibrated at test initiation, the mid -point of the test, and
test termination. Data from these systems is recovered daily and stored on
magnetic disc and hard copy. Samples of dilution water, effluent at the sampling
site, final effluent, and the receiving stream upstream and downstream of the
discharge point are analyzed for hardness es feasible. Where applicable, daily
residual chlorine measurements will be made at the above described sites.
During the on -site evaluation, Biological Monitoring Group personnel collect
benthic macroinvertebrate samples at the upstream, downstream and dilution sites
(see Benthic Macroinvertebrate Survey appendix). Where appropriate,
electrofishing is undertaken upsteam and downstream of the discharge to obtain
resident fish population data. On a site -specific basis, various other efforts
are undertaken, such as monitoring dissolved oxygen levels in the receiving
stream.
On a daily basis, test chamber screens are cleaned, effluent and dilution
pumping systems are checked and adjusted as necessary, and pH, dissolved oxygen,
and fish mortalities are recorded for each chamber. Dilution water is generally
collected on an alternating day basis, depending on need.
Two separate 24 hour composite samples of effluent are collected by means of
an automatic sampler for chemical analysis. Receiving stream and dilution water
samples are also taken for chemical testing.
Static 48 hour Daphnia pulex bioassays are conducted on a 24 hour composite
sample of the effluent and a grab sample of the influent and persistence test
aging is begun on the effluent sample.
A photographic record is made of the treatment facility, sampling points,
receiving stream, and sampling procedures while on -site.
A the end of the 96 hour test period, the dilutor is turned off and final
mortality observations are made. Breakdown and packing routines are performed
and the mobile laboratory is transported back to the Cary Aquatic Toxicology
Laboratory.
Several special care operating procedures should be mentioned. At
facilities that discharge for only a portion of the day, effluent samples are
composited by the dilutor system into a large reservoir on board the mobile
laboratory for use as the effluent while discharge is not in progress. If the
effluent has a high oxygen demand, aeration systems for the test chambers are
utilized and dissolved oxygen levels in the chambers are monitored closely in
order to prevent levels from dropping below 40% saturation at test temperatures.
In the event that actual receiving waters are deemed unfit for the test (i.e.
potentially toxic), an alternate source of dilution water is sought in the
vicinity.
-23-

CeriOdaphnia dubis Reproduction Bioassay Procedure
Aquatic Toxicology Group
N.C. Division of Environmental Management
The Ceriodaphnia aquatic bioassay is conducted to estimate the effect of an
effluent or other water sample on reproductivity. The cladoceran Ceriodaphnia
,dubia is used as the teat organism in a 7 day static renewal bioassay. A control
and 8 concentrations of effluent ranging from 0.01% to 100% are typically used.
There are 10 animals per concentration, each animal in a one ounce polystyrene
test chamber with 15 mis of solution. The test is conducted in a 25 degree
centigrade incubator with a 16 light:8 dark photoperiod.
The test is initiated with newborn animals, or neonates. Adults having
brood sacs with eggs with visible eyespots (indicating eggs are about to be
released) are isolated and checked periodically. Neonates are removed and
grouped according to time of birth. Selected groups are then composited to make
the youngest set of 90 or more neonates born within a 4 hour period. The test is
begun when the neonates are introduced into the test chambers. Temperatures must
be within 1 degree centigrade for transfer.
The animals are transferred daily to new test chambers containing freshly
mixed solutions. Chemical/physical parameters are measured twice for each batch
of solutions. The initial value is taken before the animal is introduced and the
final value after the animal has been transferred out the next day. Dissolved
oxygen of greater than 40% saturation is necessary.
The animals are fed daily. Each organism receives 0.1 ml of fermented trout
chow -yeast -alfalfa food.
As reproduction begins, only the adult is transferred to a new chamber. A
drop of concentrated nitric acid is added to the old chamber. This dills the
young so they can be easily counted under a dissecting microscope. A mean number
of young produced per adult is calculated for each concentration. Mortality of
greater than 20% in control test organisms invalidates a test.
Guidance Document_ 1985. U.S.E.P.A_ Methods for Estimating the Chronic Toxicity
of Effluents and Receiving Waters to Freshwater Organisms. (EPA-600/4-85-014).

Benthic Macroinvertebrate Procedure
Biological Monitoring Group
N.C. Division of Environmental Management
The sampling methodology requires that a stream or river be wadable. High
water conditions may severely impair sampling efficiency by making critical
habitats inaccessible. A fixed number of samples are collected for each station.
These include: 2 kick net samples of riffle and snag areas; 3 sweep net samples
of bank areas and macrophyte beds; 2 fine mesh waandown samples of search of
rocks, Togs, leaves, and substrate The benthic macroinvertebretes are picked
out with tweezers and preserved in alcohol. A collection card is filled out
which includes such data as canopy cover, dissolved oxygen, pH, stream
temperature, substrate composition and stream morphology at the site. Organisms
are identified to the lowest possible taxonomic level, generally to species.
Density of each taxon is rated as Rare (1 or 2 individuals), Common (3 to 9),
Abundant (10 or more). Most organisms may be identified using only a dissecting
microscope, but Oligochaeta and Chironomidae must be mounted and identified with
a compound microscope. Reference collections are maintained and all samples are
kept and stored by study area.
The first level of analysis summarized the data by total number of taxa or
"taxa richness" (S) and density (N) for each station.
The second .level -o$.analysis summarized data by taxonomic groups (mostly
orders of insects). The EPT value is the sum of the taxa richness for the
intolerant insect groups Ephemeroptera, Plecoptera, and Trichoptera.
The final step in data analysis is to summarize the data separately for each
taxa. The presence of absence of individuals of "indicator species" is, in
itself, insufficient for characterizing water quality. Tolerant species will
usually become dominant only in polluted systems. Information on pollution
tolerance of any given taxon, combined with quantitative data on its distribution
can often be related to specific chemical or physical changes in the environment.
—25—

List of Definitions
Aquatic Toxicology Group
N. C. Division of Environmental Management
Acclimation - refers to the process of gradually adjusting organisms from water
of one type to another so that the organisms are not stressed from
radical changes in temperature, hardness, pH, ionic strength, etc.
Acute toxicity - the effect a short term exposure to a chemical or substance has
on an organism; usually defined as death of that organism.
Application factor - a value which estimates an instream toxicant level that
will be safe at a chronic level for resident organisms from acute
toxicity data, usually defined by a fraction of the LC50.
Aquatic - having to do with water.
Aquatic Toxicology Group - the group within the Biological Services Unit (Water
Duality Section) which performs aquatic bioassays for the Division of
Environmental Management. The Group is located at the Cary laboratory
facilities. All test organisms (including Daphnia pulex, Ceriodaphnia
sp., and fathead minnows) are cultured at these facilities by Aquatic
Toxicology personnel.
Benthos/Benthic macroinvertebrates - a wide assemblage of invertebrate animals
(insects, crustaceans, molluscs, etc.) which live in streams, are an
important food source for fish populations, and are used as long term
water quality indicators.
Bioassay - a test used to determine the effects of a chemical or substance on an
organism.
Cadmium - one of the toxicants recommended by EPA for quality assurance testing
of the health of aquatic organisms.
Calibration - the adjustment of meters or systems with standards of known values
in order to assure the quality of data obtained from these meters or
systems.
Ceriodaphnia sue. - a small cladoceran crustacean. It is found throughout most of
North America and obtains a maximum size of approximately 1 mm. This
organism has been adopted for aquatic bioassay testing because of its
small size, ease of culture under laboratory conditions, stability of
genetic strains, and sensitivity to toxic substances. It is generally
used in a 7 day static renewal "mini -chronic" bioassay testing for
mortality, time to sexual maturity and reproductive rate. Ceriodaphnia
sp. is accepted in the field of aquatic toxicology for testing in
moderately soft waters.
Chronic toxicity - the effect of a chemical or substance on an organism, usually
during a longer period of time than that measured for acute toxicity.
This effect is usually measured as a non -fatal response (eg. reduction in
growth, egg production, predator avoidance, feeding rate, etc.). Test for
chronic toxicity are frequently performed during the entire life cycle of
the organism.
Composite - a sample or method of sampling used to obtain data on a substance
which may vary over time or space. For example, a time or temporal
composite of a stream would be one collected at intervals of time at the
same location. This is frequently accomplished with automatic sampling
devices.
-26-

Daphnia pule), (water flea) - a small cladoceldn Crustacean. It is tuunc
throughout most of North America and obtains a maximum size of
approximately 3.5 mm. This organism has been adopted for aquatic bioassay
testing because of its small size, ease of culture under laboratory
conditions, stability of genetic strains, and sensitivity to toxic
substances. It is generally used in a 48 hour static bioassay testing for
mortality. D. pulex is widely accepted in the field of aquatic toxicology
for testing in moderately soft waters.
Design flow - the volume of water and waste that is initially planned to pass
through a facility or waste treatment plant and still allow maximum
operating efficiency. Design flow is usually expressed in millions of
gallons per day (mgd).
Dilution (water) - the water used in bioassay tests to dilute the waste water to
various concentrations (expressed as percent). Wherever possible, this
water is from the actual stream that receives the waste, upstream from
that waste. When this is not possible, other suitable water is obtained
Dilutor - refers to a modified Mount and Brungs design serial dilution apparatus
which receives dilution water and effluent/waste and, through a series of
chambers and electrical solenoid valves, mixes the effluent and dilution
into a series of concentrations for the test (expressed as percentages of
100% effluent).
Electrofishing - method for collecting fish using electrical shock to
momentarily stun the fish so they float to the surface and are easily
netted.
Effluent - the waste water exiting a facility which is discharged as treated
waste to a stream or as untreated waste to some other facility.
Fathead minnow (Pimephelas promelas) - a small fish which occurs throughout much
of North America. It obtains a maximum size of approximately 100 mm and
is raised commercially as bait fish. The fathead minnow has been raised
for numerous generations in a number of laboratory culture for use in
toxicity testing. The fish can produce eggs year round in the laboratory
environment under correct conditions which produce test organisms as
needed.
Flow -through - the flow -through bioassay utilizes which either continuously of
occasionally replace effluent/toxicant concentrations throughout the test
in an attempt to simulate stream conditions where new effluent and
dilution water ere continually flowing through an organism's habitat.
Hydrolabw - a multiparameter instrument which measures and records temperature,
pH, dissolved oxygen, and specific conductance of water.
Instream waste concentration (IWC) - the percent concentration of an
effluent/toxicant which is present in a stream under assumed worst case
conditions. The IWC is derived from the formula: [DF / (7Q10 ; DF)] x 100
= IWC CO, where DF is the design flow (in cfs) of the facility in
question and 7Q10 is the 10 year, 7 day, low flow (in cfs) of the
receiving stream.
LCSO - that concentration or percentage of a waste/chemical/substance which is
lethal to 50% of test organisms over a stated period of time.
"Use of this tern or system does not constitute an endorsement
-27-

NPDES - National Pollutant Discharge Elimination System. A system devised by thF
Federal Government and adopted by North Carolina for the permitting,
monitoring, and pollution abatement of dischargers to surface waters.
Neonate - roughly translated to newly born. In reference to Daphnia pulex, the
neonate refers to the life stage in the first and early second instar,
generally the first 24 hours of its life.
Screening bioassay - a testing system established to determine general levels of
acute toxicity of compounds/discharges using 48 hour Daphnia pulQx tests.
7Q10 - the measurement of a stream's lowest average daily flow over a 7 day
period during a 10 year span, generally stated as flow in cubic feet per
second (cfs).
Sodium pentachlorophenate - a chemical accepted by EPA as a toxicant for quality
assurance testing of the health of aquatic organisms. This chemical is an
organic pesticide.
Static - refers to an aquatic bioassay in which toxicant/effluent concentrations
are set up at the beginning of the test and not changed for the rest of
the test. This test is generally short term as compared to a flow -through
or replacement test because of potential degradation of the
toxicant/effluent.
Taxa - refers to a group of genetically related organisms, (i. e. genus, order,
species).
Taxa richness - number of taxa.
30Q2 - the measurement of a stream's lowest average daily flow over a 30 day
period during a 2 year span, generally stated as flow in cubic feet per
second (cfs).
Toxicity - the adverse effect of a chemical/substance on an organism. Toxicity
is usually defined as a fatal or non -fatal response over a give period of
time.
UT - Unnamed tributary - a term given to streams which have no accepted name.
-28-