NC0005177_REPORT_19880610

i
NPDES DOCUWENT SCANNINS COVER SHEET
NPDES Permit: NC0005177
FMC Lithium
Document Type: Permit Issuance
Wasteload Allocation
Authorization to Construct (AtC)
Permit Modification
Speculative Limits
Report
Instream Assessment (67B)
Environmental Assessment (EA)
Permit
History
Document Date: June 10, 1988
Thi"document Jim printed on revise paper-ignore a ny
content on the reYeree Side

DIVISION OF ENVIRONMENTAL MANAGEMENT
June 10, 1988
M E M O R A N D U M
TO: Ron McMillan
Dennis Ramsey
FROM: Steve W. Tedder /Y
SUBJECT: Toxicological. Evaluation - Lithium Corporation of America
Attached is the final report concerning an intensive toxicological eval-
uation of the Lithium Corporation of America in Gaston County
If there are any questions, please contact myself or Ken Eagleson at
[phone].
SWT:ps
cc: Ken Eagleson
Larry Ausley
Bob DeWeese
Trevor Clements
Jay Sauber
Jim Overton
Central Files

Lithium Corp.
NPDES NC0005177
On-Site Toxicity Evaluation
]I1111111111 I11111111111
North Carolina Department of Natural
Resources & Community Development
MOBILE 19&
Bioassay and Biomonitoring �� o
LABORATORY
6•
mo o. "> 'f,'�.� roc
0 0
o90 oao
0 0
NORTH CAROLINA DEPARTMENT OF NATURAL
RESOURCES AND COMMUNITY
DEVELOPMENT
WATER QUALITY SECTION
June, 1988

LITHIUM CORPORATION OF AMERICA
TOXICITY EXAMINATION
NPDES NO. NCO005177
NORTH CAROLINA DEPARTMENT OF NATURAL RESOURCES
AND COMMUNITY DEVELOPMENT
Division of Environmental Management
Water Quality Section
.Tune 1988

TABLE OF CONTENTS
Page
Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
ToxicityExamination. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Chemical Sampling Analysis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Benthic Macroinvertebrate Community Analysis. . . . . . . . . . . . . . . . . . . . . . . . . . ... . . . 14
Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Recommendations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Footnotes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Appendix
Species List-Benthic Macroinvertebrate Study. . . . . . . . . . . . . . . . . . . . . . . . . . 27
Ceriodaphnia dubia Test Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
96 Hour Flow-through Test Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Ceriodaphnia Reproduction Test Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Benthic Macroinvertebrate Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
List of Definitions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
LIST OF FIGURES
Figure 1. Seven Day Ceriodaphnia Mean Cumulative .Reproduction. . . . . . . . . . . . . 5
Figure 2. Study Area and Sampling Sites, Lithium Corporation. . . . . . . . . . . . . . 8
LIST OF TABLES
Table 1. Self-monitoring Toxicity Test Results. . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Table 2. 96 Hour Fathead Minnow Mortality. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Table 3. Sampling Station Descriptions - Lithium Corporation. . . . . . . . . . . . . 7
Table 4. Results of Chemical. Analyses. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Table 5. Station Descriptions for Macroinvertebrate Sampling. . . . . . . . . . . . . 17
Table 6. Benthic Macroinvertebrate Taxa Richness, by Group. . . . . . . . . . . . . . . 19
Table 7. Summary Statistics, Lithium Corporation Study. . . . . . . . . . . . . . . . . . . 20
Table 8. Statistical Tests (Wilcoxon Signed-Rank Test). . . . . . . . . . . . . . . . . . . 21

INTRODUCTION
An Intensive on-site toxicological evaluation was conducted at the Lithium
Corporation of America (NPDES #NC0005177) from September 21 through 26, 1987.
Lithium Corporation processes lithium ore and produces more than 70 lithium based
products. The plant has been operating since 1941. Industrial process waste-
water (0.595 MGD flow) generated from the lithium processing and production
plant, along with treated domestic wastewater (0.005 MGD flow) from one of two
package treatment plants at the Lithium facility, are discharged into the indus-
trial waste treatment lagoon and make up the discharge of pipe 001. A second
discharge pipe (002) consisting of treated domestic wastewater (0.015 MGD flow)
from the second package treatment plant was not tested during this evaluation.
Effluents from both discharge pipes flow into an unnamed tributary (UT) to Aber-
nathy Creek, however the 002 discharge is upstream of the 001 discharge.
This report contains findings of toxicological and chemical evaluations
performed, including the. following: '
1) 48 hour static Ceriodaphnia dubia toxicity tests on wastewater treatment
lagoon influent and effluent to determine acute toxicity;
2) 96 hour static toxicity test using Pimephales promelas (fathead minnows)
on effluent to determine acute toxicity;
3) Seven day Ceriodaphnia dubia static replacement reproduction suppression
toxicity test on effluent to determine acute and chronic toxicity;
4) 48 hour fractionation toxicity test on effluent to identify or rule out
classes of constituents causing acute toxicity;
5) Analysis of chemical samples collected from the wastewater treatment
system influent and effluent and receiving stream above and below the discharge.
The process wastewater is first treated with sulfuric acid to adjust the pH
to between 6 and 9. The wastewater then flows through a pipe and ditch to a

waste (tailings) lagoon with a holding capacity of approximately 100 million
gallons. The effluent then flows through a flow measurement weir into the
unnamed tributary (UT) to Abernathy Creek. This stream is designated as a "Class
C" stream in the Catawba River basin with a 7 day, 10 year low flow (7Q10) of
0.20 cubic feet per second (CFS). The permitted effluent flow for the process
wastewater discharge is 0.600 million gallons per day (MGD), giving an instream
waste concentration (IWC) of the effluent in the UT in Abernathy Creek of 82.3%,
during 7Q10 conditions.
TOXICITY EXAMINATION
The intensive on-site toxicological evaluation was conducted at the Lithium
Corporation of America as a result of toxicity detected in tests performed by the
Division of Environmental Management (DEM) :
Test Date Test Type Result
860508 48 hr Daphnia pulex LC50=15%
870416 48 hr Ceriodaphnia dubia LC50=29%
870528 " it1 11 ilLC50=26%
An LC50 is the concentration of effluent lethal to 50% of the test organ-
isms.
On July 1, 1987, the DEM instituted, by Administrative Letter, a self-
monitoring requirement for Lithium Corporation to perform the 48 hour static
acute toxicity test on a monthly basis.
Lithium Corporation was requested by DEM on August 6, 1987 to submit an
evaluation report dealing with proposed actions to reduce the demonstrated
toxicity of their wastewater. The first 6-month report was submitted by Lithium
Corporation February 4, 1988 and is in review. A chronic toxicity limit was
included in the NPDES permit re-issued to Lithium Corporation on February 4,
1988, specifying that the company perform monthly Ceriodaphnia reproduction
Pass/Fail tests at an effluent concentration of 83% as a monitoring requirement
only, until February 28, 1989. On March 1, 1989, the requirement will change to
-2-

quarterly monitoring, but the 83% effluent must pass the test as a whole effluent
toxicity limit. Self-monitoring results reported to date are presented in Table
1.
Table 1. Self-monitoring Acute Toxicity Test Results for Lithium
Corporation.
Test Date Test Type LC50
July, 1987 48 hour Daphnia up lex Bad test
August, 1987 48 hour Daphnia pulex 6.5%
September, 1987 48 hour Daphnia pulex Bad test
October, 1987 48 hour Daphnia pulex Not reported
November, 1987 48 hour Daphnia up lex 20.7%
December, 1987 48 hour Ceriodaphnia dubia 47.2%
January, 1988 48 hour Ceriodaphnia dubia 54.9%
Four bioassays were conducted during the on-site toxicological evaluation: a
96 hour flow-through bioassay using Pimephales promelas (fathead minnows), two
Ceriodaphnia dubia 48 hour static bioassays, and a 7 day static replacement
reproduction suppression bioassay using the cladoceran C. dubia. Final effluent
samples were collected at the discharge box prior to the V-notch weir. Dilution
water was obtained from Beason Creek at SR-2246 (Cleveland County) . This dilu-
tion water was tested prior to use in the on-site investigation with the C. dubia
reproduction suppression bioassay and yielded reproduction similar to that in
Aquatic Toxicology Laboratory cladoceran culture water.
The fathead minnows used in the 96 hour bioassay were cultured at the
Aquatic Toxicology Laboratory and were 57 days old at test initiation. The min-
nows were acclimated to water from Beason Creek on September 16-18, 1987. Ten
fish were transferred randomly to each test chamber 12 hours prior to test ini-
tiation. Replicates at six effluent concentrations (5, 10, 25, 50, 75 and 100%)
and control/dilution water were tested. The test was conducted from 8:37 AM,
September 22 to 8:37 AM, September 26, 1987. It should be noted that for the
first 56 hours the test was conducted under flow-through conditions. Management
-3-

personnel at Lithium decided to stop discharge of effluent to the UT to Abernathy
Creek due to an effluent pH level detected approaching their high end limits.
The fathead minnow test was conducted under static conditions from the 56 hour
through 96 hour period. Mortality in this bioassay is summarized in Table 2.
Trimmed Spearman-Karber (TSK)1 analysis of this data results in an LC50 of 70%.
Table 2. 96 Hour Fathead Minnow Mortality
Effluent
Concentration W Mortality (7)
0 0
5 0
10 0
25 0
50 5
75 60
100 95
Ceriodaphnia dubia 48 hour static bioassays were conducted while on-site
using a 24 hour composite effluent sample and an instantaneous grab sample of the
influent. The resulting LC50's of 0.32% for the influent test and 21% for the
effluent test indicate that there is partial removal of toxicity by the waste
(treatment) lagoon.
A seven-day static replacement Ceriodaphnia dubia reproduction suppression
bioassay was performed from September 21 through September 28, 1987. Mortality
was calculated using the TSK method and resulted in a 158 hour (7 day) LC50 of
11% for C. dubia. Ceriodaphnia exposed to effluent concentrations up to and
including 1.0% had similar mean overall reproduction and survival when compared
to control organisms. No young (neonates) were produced by females exposed to
25Z, 50%, 82.7% (facility's IWC), and 100% effluent. An average of 10.1 young
were produced per female exposed to 10% effluent compared to an average of 33.2
young per female exposed to 1% effluent. Mean cumulative reproduction is
depicted in Figure I. The No Observed Effect Concentration (NOEC) on reproduction
-4-

Figure 1 . Seven Day Mean Cumulative Reproduction
40
-a- Control
-t 0.1%
-CF 1.0%
o- 10%
30
Mean Young
Produced
20
No reproduction
occurred in the
25-100%effluent
concentrations
10
0
1 2 3 4 5 B 7 8
Day of Test
Lithium Corporation
-5-

was 1%, with a Lowest Observed Effect Concentration (LOEC) of 10%. A Chronic
Value of 3.2% was calculated from this data as defined in EPA document
600/4-85/0142.
In addition to the on-site bioassays, an effluent characterization, or frac-
tionation test (as described in the 1987 EPA draft document "Methods for Toxicity
Reduction Evaluations, Phase 1, Toxicity Characterization Procedures") ,was con-
ducted at the Cary Laboratory using Ceriodaphnia dubia as the test organism.
Effluent characterization is a way to determine the physical/chemical constitu-
ents of an effluent that are contributing to toxicity. By conducting a series of
tests the effluent can be screened for specific groups of toxicants, such as met-
als, volatile organics, oxidants (i.e. chlorine), etc. . Each characterization
test is designed to eliminate or reduce the biological availability of a particu-
lar toxicant, through chemical or physical treatment. The fractionation tests
conducted on a grab sample of Lithium Corporation effluent collected May 27, 1987
are listed below:
- Degradation Test
- Filtration Test
- Air Stripping Test
- Oxidant Reduction Test
- EDTA Chelation Test
- Solid Phase Extraction Test
- Chloride Removal Test
The ET50 (ET50--nedian lethal time, this is the end-point used in timed
lethality tests) of the untreated effluent sample was 6.0 hours. Results of the
Air Stripping test indicated significant increases in toxicity occurred in the
aerated neutral and aerated acidified fractions, with ET50 values of 2.0 and 1.6
hours, respectively. None of the standard fractionation tests conducted
resulted in a reduction in toxicity.
The nthiium Corporation effluent is-known:-to-contain-hi-gh--enough l-evels-o.
chloride-(effluent-cant: .8.7.0924a4.9.0-mg/_l.,_87.0.926!E540-mg/_1)`t"o�suspect=i.t as a
(possible-toxicant. The chloride concentration was measured as 850 mg/1 in the
-6-

grab sample collected May 27, 1987 for use in the characterization/fractionation
tests. Total residual chlorine was also measured, with a concentration of 0.2
ppm recorded. Two aliquots of this sample were passed through an ion exchange
column to remove the chloride. The average ET50 of the column fractions was
12.25 hours, a value more than twice that of the baseline (untreated) effluent.
This represents a significant reduction in toxicity. It was concluded that the
presence of chloride contributed to the effluent's toxicity.
CHEMICAL SAMPLING ANALYSIS
Samples of final effluent, influent, toxicity test dilution water, and the
receiving stream above and below the discharge were collected for chemical analy-
sis on two dates during the on-site evaluation. These samples were analyzed at
the Division of Environmental Management chemistry laboratory. Sampling stations
are described in Tables 3 & 5 and presented schematically in Figure 2. All
samples were collected as instantaneous grabs with the exception of Station 02
(effluent toxicity test sampling point) which was sampled as a 24 hour compos-
ite. It should be noted that the 24 hour composite effluent sample of September
26 was collected in the waste lagoon because Lithium Corporation personnel had
shut off effluent flow to the discharge box (previously described above) .
Results of chemical analyses are given in Table 4.
Table 3. Sampling Station Descriptions
Station Location
01 Abernathy Creek @ I-85 approximately 100 yards
above point where UT to Abernathy enters
02 Final effluent @ discharge box prior to V-notch weir
02A Influent to waste lagoon @ pipe outfall on upper end
of waste lagoon
03 Abernathy Greek @ SR-1302
04 Beason Creek @ SR-2246
-7-

Figure 2. Study area for Lithium Corporation, Gaston County.
N
0
BESSEMER
CITY
UT ABERNAT\CRLITHIUM
CORP
� 02A
BEASON CREEK,
STATION 04, TAILINGS
POND I-85
10 MILES EAST
^� SR 1302
G
US 29
LIT ABERNATHY CR
03A
ABERNATHY CREEK
KINGS MOUNTAIN 01
03
ABERNATHY CREEK
1-85
yu'Y�;fr
r.Y
1 MILE
L_ i

Table 4. Chemical Analyses Results-Lithium Corp- of America
Permitted Flow--( 0,6!
..................... ........... .................... .....................
.......................(W) I
......................... ......................... ............................................................ .................... .....................
7QIO (CFS) 0.
li-d......... ..................... .......... ...................... .............. .......................................
........................................... ..................... ................................................................................................. ............... ...................
.............................................. ................­­........................a.........---.........y...................... ................ ........................................
Chemical/P�V�ical Units Water Qua). Sta 01 Sta 02 Sta 02A Sta 03 Sta 04
.......... ..................­1................................................................................... ................­­.............
Standards '709'4 870924 870924 870924 870924
Analyses .......................... . __
....... .�s 0. t-
........ .......... ................. ........................ ........... ...........................................
............. .............. ....... ........ ............. ..........
BOD PPM
............. ...... ....................4.......................................... ...... ...............................
COD = PPM <5 69 83 11 5
............................................................. ..................................... ....................... .................... ......................
2700 3800 f 100 72
Re_-;due TOTAL PPM b
.................................................. ........................ ....................... ..................... ...................... ................. .............................................
volatile PPM 40 170 690 62 42
............ ....................... ................... .................*....... ......................................................................................................
fixed PPM 28 2600 3100 990 30
.... .......................... ...............::
................... ............... .................. ...... ............... ....................
Residue SUSPENDED PPM 3 11 28 5
.......................................... .................. -----------------------................... ......................................... ...........................................
I I
volatile PPM 3 r.
............................................................................................................... ...............
�1­1_1_111­1.......4.......................>.............................................
fixed PPM 22 4 2
........................................................................ .............................................................................. ­­­, .......... ......................
7
pH (standard units' 6.0-9.0 7.4 C.6 6.3
................................. ........... ...... .................... ............ ...........................................
PPM 1
............. ......................................................... .................. ................................. ------------- ......................
Alkaliniti�.................... PPM 27 200 241D 83- 18
............ ............................ .......... ................... ............
PPM
.....................................
.......... ...................... ............................. ............. .................... ......................
Hardness PPM -4 1 120 47 70J 14
.............-........................... .................................................................................. ............ .......................................... ......................
1 4100 4100 1700 6.3
............ ..................... ... ...................................
, oifio Conductance :uMhos 'on-,: 70
.............. ..................... ............
NH3 PPM 0.m, <m-.11 �0.01 0.04 -"0.0 1
....... ................................. ............. .............;..................... ...................... ......:................
.............. .................. ...........
TKN PPM 12 0 0,8 0.4 0.3 0.1
...................... ................................... ............... .....................I...............
NO2 NO3 PPM 0.21 0.03 0.46 0.15 0.50.
........................­­­........................................... ................................... ............... ...... .......... .................... .......
P. folal PPM 0.04 0.06 0.1 0.04 0.02
................................................................. ......................................................... ........................................... .............. ......................
I Of AOT 1-7-5 <25 05 <25 <25
Silver PPB : I-
.........................­­­........... ..............................
------------- ................. 4 ................... ..........
200 200 2200 950 200
Aluminum PPS
........................................................................ ................................................. .............................................. .....................I .............
Cadmium PPB 2 <1 ID <10 '10 <10 '10
..................................... ..
................................ ................. ..................... ..........................................................................................
Chrorniurn (Total) PPE?* 50 <25 ..25 <25 <25
................................................................................................................... .................... ................... ...........
PPI? I 5(ALY 1:10 <10 19 11 <10
....................... ....................... ........... ...................... ................ ......... .......................
1200 7300 270
Iron PPB 1000 41 C1 140 -
.......................... ..................................... ...................I.............. .......................................... .......... .................... ......................
t 12
0.21, <0.`� <0.2 Merour.y .......................... PPB j
.................. ...................................... ...... ........................
<25 30 75 : 05
.............. ........................................................................................................ .....­-..- f�< C,
INickel PPB 50 <50 <50 <50 5 <50
........................ ......­­­­­.:.................................... .............. ...................... ...............11111­�
..........................................
Lead PPB --)5 ZO <5O <50 <50 <50
............................................. ........................ ............. ............. ..................... .......... ....................I.......................
Zinc PPB
50(AL) 16 28 29 <jo 'jo
.................................................. .................................. -------------------- ---------------------- ........................................................
Berq Ilium PPB 11 <25 <25 <25 <25 <25
..................... ..................... ....................................__.............................................................. .............. ..................
Lithium PPS
.................................... ............................................................ ................... ............................................................
PPB 10 <9 <20 :: <=
2 <20 <5
?.................... ......................
........................................... ............................................ ................................................................
............
Values..rep.�!�!nt action levels as specified in -021 1(b)(4)
...............................specified...................... .............. ....... ­------------- ......-.......
.... ........................ 'Fresh...'dater Classifications Standards
.......................... ........................ ............ --------------------- ...................... ....................... ................ .....................
....................... .......................... -...................................-.............................I.......... ...................... .. .............
-9-

Table 4. Chemioal Analyses Results-Lithium Corp- of Amerioa
Permitted Flow(MOD) 0.6;
................ ........ ...... .............. ........... ........ ........................................................
..............
,.....'
................ ............ ..6....2..3.............................................. .................
..........................................................................................
7010 (CFS) 0.2:;
.................................... ...... .............................. ................... ----------- -----------_--- _---------_-------
......................................................................................................................... ............................................................... ..................................................
...........I...... .......................................... ?..................... ....... ................. ....................................................
UnemicaUHhUsioal Units ISta fj I LS't a 02 Sta 02A : Sta 03 Sta 04 Predicted stream**
..............I..................... ......................... ............ ..... .................. ....................................*.............870926 S70926 870926 iP.370926- 0926 cono. at 7Q1 0
.......... ................ ..................... .................... .......... ..................... .........................................
........................................................................................................................ .................... ..................................................................................................
BOD PPM
............. ...... ..................... ................................... ..................... ...................................................
COD PPM 5 63 45 5 5 54................................................................................................. ..................... .................................................................................................2 Residue TOTAL PPM 130 2800 1300 300 41 2274
............... ..................... ........................................I................... ............... ...........................................
volatile PPM 72 120 311)0 43 10
........................ ....... ..................... ......... ......................
fixed PPM 100 2700 960 250 31
................................................... ................. ------- ..................................... :................ ............... ........Residue SUSPENDED = PPM 9 27 11 3 3 15.7
--------------*............................................................. .......... .........................................I..........................................................
7 .2� 3
yolatile PPM 6 10
............................................. ....................... ..................... ............
fixed PPM 3 17 4 1 <I
........................................................................................................................ ...................................................................................................
PH [standard units) 8 7.4 7.2
7.8 9.3 ::
....................... ------ .............................................................. ..................... ........................................................
AciditqPPM 3 40 6.............................................................................................:.......................................... ...................
.......................................................................
.... ........AI.ka.Ij.n#y.............................;.......PPM.:'.......PPI-I 44 200 :� 240 :' 49 13 165
......­.- . .............. ............................................................. ........ ................. .............................
Chloride PPFI 5 540 560 33 4 426
....................... ................. ........... ...................................................... .................... .....................................................................................................
Hardness PPM .20 120 63 14 99
................................................... ------------------------ ........................................... .......... ........ .......... ...................
Specific Conductance uMhos/cm 170 4200 2100 510 640 3432
.......... ...... ............ ....................................
NH3 PPM 0.46 0.04 0_05 0.47 13.07, 0_0 7_%
................................................... ........................ .................... . ............................................... ..................... ...............................
TKN PPM 0.6. 0.6 0.3- A 7 i <O.1 0.5
................................................................... .................................................... .................... ...... ...............................................................................
NO2 NO3 PPM 2'.1 1 0.03 0.5 1.7 0.58 t: 0.0.2
.......... ........................................ 1�
........................................... ..................... ......................... ..........-......
P. total PPM 0.19 0.0-1 02 OA6 0.02 ........................
............I................................ .................................................................... .................... .................................... .........................
Silver PPB <25 <25 <25 <25 <25 <25
.................................................................... ---------------- .............. ............ ............................ ........................... ................
Aluminum ppe 1100 300 6.50 750 100 20.
........... ........ ................. ..............­........................... ................................................................................
Cadmium PPB <10 <10 :: '.10 �: '113 <10 110
.................I................................ ........................ ..................... ..................... .................................... .................... ...................................................
05 <25 <25 <25 <25
Chromium(Total) PPB <2� �!' 4
............................................ .....................................: .............................;.........................................................................................................................
1 -.10 1 1 f,
PPE. I I �10 0
....... ........................................ .............................................................................................................. ..................... .........................................................
Iron PPB 800 :: 180 1100 440 190
.........................................................................................................................................
........................... ..................... ...............
2 M�_rout PPS <O.� KO.2 1 <02 <0 2 <0.2'
............... ....................... ..................... ................................... .......... ................................................-.........................
M.9 PPB 3 5 40 85.......................................................................................................... ............ ................................. ................
nickel PPB <50 <50 (50 <50 <50 <50
................................................... ........................ ..................... ............................................................... ...................................................................
Lead PPB <50 <50 (50 (50 <50 <50
............ .......... ......................t.................... .................................. ...................
............................................ ..
7inC;
I PPB <1011 32 <10 <10 16A
................................................... ........................ ........<25......... .............. ................................. ...................
Berulliurf? PPB �: :� <25 <25 <25 <25 2_5
............................................................. ........................................ ................
Lithium PPB <25 85000 100000: 4000 <25 85000
::
.............:..................................... ........................ ..................... .......................................... .................. ..................... ........ ............
Selenium PPB <5 <20 <5 <20 <20
..............................­1............ ................. ............z.................................................................................................
...................................................*..."...................................................................................................
_ -
q Values represent predicted instream concentratlons.��!!!q..averae effluent
................................... ..................... ............................................................................................ ................ ..................1,................................
concentrations and assuming upstream concentrations of 0-
.......................................................upstream 1.....................--------------------- ............................................................................
....................................................................... ...........................................
.......................................................................................
............*.......
_10-

Metals analyses revealed that small quantities of copper were being dis-
charged into the waste lagoon with concentrations of 19 ppb and 12 ppb in the
influent (02A) samples of September 24 and 26, 1987 respectively. No copper was
detected (detection limit 10 ppb) in the effluent collected September 24 and 26,
or in dilution water (04) samples. Zinc was detected in the effluent on both
sampling dates, with 28 ppb on September 24 and 11 ppb on September 26. At these
i
concentrations and 7Q10 conditions, the concentration in the receiving stream
would average 16. 1 ppb, assuming no background levels. Zinc was found upstream
(01) in the September 24 sample only, at a concentration of 16 ppb. No zinc was
detected in downstream (03) or dilution water (04) samples collected on either
date.
Lithium was_detected in--the-influent(02A)—effluent(02)-,-and_downstream
(-03) samples-collected-September-26-at-concentrations of 100.,004_ppb,_85,000 ppb?
F,'and-4000-ppb-f.or-each-site-respect-i-vely7 No lithium was found (detection limit
<25 ppb) at either the upstream (01) or dilution water (04) sites. There is no
S•
additional effluent lithium concentration data available (other than a concentra-
tion of 60,000 ppb detected in a 24 hour composite sample collected by DEM per-
sonnel on May 7, 1986) because the NPDES permit issued to Lithium Corporation
does not specify monitoring of effluent lithium concentrations. Evidence-of
L-ith"ium Corp oration'''s-discharge.was detected_in-Abernathy-Creek-downstream-of_the
confluence-of-the-UT-to"A'bernathy Creek;-instream_concentrations_of-1ithieim+
increased-dramatically from-<2.5-ppb"(-16wer detection_limit.)_to.-4000_ppb-i
samples collected-at-the-upstream-and 'downstream-sites respectively.
As discussed previously chloride was detected in final effluent samples col-
lected September 24 and 26 with concentrations of 490 ppm and 540 ppm recorded
for each day respectively. A comparison of chloride concentrations in upstream
vs. downstream samples indicate the effluent increased chloride levels in Aber-
-11-

nathy Creek. On September 24, upstream and downstream concentrations were 2 ppm
and 170 ppm, respectively; and on September 26 concentrations increased from 5
ppm to 33 ppm.
Zinc LC50's have been reported as low as 76 ppb4 in C. reticulata 48 hour
bioassays. Fathead minnows are less sensitive to this metal than Ceriodaphnia,
with a 96 hour LC50 of 600 ppb5.
Toxicity data on lithium in the peer-reviewed literature is sparse, however
in a 64 hour Daphnia magna test done with lithium chloride the FC50 was reported
as 1200 ppb6.
In toxicity tests conducted with sodium chloride as the test compound chlo-
ride LC50' s have been reported as low as 1413 ppm7 and 1470 ppm8 in Ceriodaphnia
and Daphnia pulex 48 hour acute bioassays respectively, and a Chronic Value of
372 ppm for D. pulex has been reported8. Fathead minnows are slightly more
resistant to chloride than these cladocerans with 96 hour LC50's reported as low
as 1830 ppm9.
A comparison of the Lithium chloride and sodium chloride toxicity data sug-
gests that lithium is a significantly more toxic substance (by more than 3 orders
of magnitude) than chloride, at least in terms of Daphnids. Because no lithium
toxicity data is available for fathead minnows the presence of lithium in waste-
water discharged from Lithium Corporation's lagoon can not be definitely impli-
cated as a causitive agent of the recorded mortality in the on-site 96 hour fath-
ead minnow test. However, in light of the elevated effluent lithium concentra-
\.ltion detected it can not be ruled out.
Total residual chlorine (TRC) levels were measured in the final effluent
during the on-site evaluation. Chlorine was monitored because of its potentially
toxic effects instream. All morning TRC analyses were run on an aliquot of the
effluent sample used to set up the daily replacement solutions for the 7-day
Ceriodaphnia reproduction bioassay. Final effluent TRC (in ppm) detected while
-12-

on-site are presented below (M=morning measurement; A=afternoon measurement;
NM=not measured):
Mondav Tuesday Wednesday Thursday Friday Saturday
M M A M A M A M A M
NM 0.06 0.09 0.08 0.06 0.07 0.04 NM 0.05 0.08
The average TRC for the morning measurements was 0.07 ppm.
Forty-eight hour LC50's for chlorine have been reported as low as 0.017 ppm
for the cladoceran Daphnia magnalo. Fathead minnow 96 hour LC50's have been
reported as low as 0.082 ppm10 in a flow-through system. Based on this toxicity
data, concentrations of effluent chlorine may have contributed to the mortality
recorded in the 7-day Ceriodaphnia test and 96 hour fathead minnow test. The
influent to the waste lagoon was measured for TRC on September 23 and 24, 1987
with values of 681 ppm and 147 ppm recorded on each day respectively. A compari-
son of influent TRC vs. effluent TRC indicates that the waste lagoon signifi-
cantly reduced TRC concentration arkd acute toxicity of the process wastewater
(assuming that influent TRC concentrations remain elevated over time) , however
the final effluent was still acutely toxic to Ceriodaphnia and fathead minnows.
All chemistry sampling stations were checked twice while on-site to assess to
what extent, if any, Lithium Corporation's wastewater contributed TRC to the
receiving stream. Analysis conducted on September 23 revealed an increase in `SRC
from 0.03 ppm to 0.12 ppm at the upstream and downstream sites respectively. An
increase was also detected on September 24 with 0.02 ppm and 0.06 ppm recorded at
the upstream and downstream stations respectively.
Organic analyses were conducted on influent and effluent samples collected
September 24 and 26, 1987. Only one unidentified peak was detected at a concen-
tration of 20 ppb in the September 24 influent sample. It appears that organic
chemicals were probably not responsible for the observed effluent toxicity.
-13-

BENTHIC MACROINVERTEBRATE COMMUNITY ANALYSIS
The Benthic macroinvertebrate community in the receiving stream was sampled
to detect any adverse effects below the Lithium Corporation discharge. Samples
were collected on June 10, 1987, at one site on UT to Abernathy Creek below the
Lithium discharge and two sites on Abernathy Creek, above and below its conflu-
ence with the UT. In addition, the toxicity test dilution water collection site
was sampled.
Benthic macroinvertebrates were collected using DEM's standardized qualita-
tive sampling method. This method uses a wide variety of collection techniques
to inventory the aquatic fauna present in all habitats within the stream. The
primary output of data is a list of species (or taxa) collected with an indica-
tion of relative abundance (Rare - 1 or 2 individuals, Common - 3-9, Abundant -
10 or more) for each taxa. Benthic macroinvertebrate communities of unstressed
streams and rivers have many taxa present, while the communities in polluted
areas have fewer taxa. The total number of taxa present or "taxa richness" (ST)
5
and the number of taxa present from the pollution intolerant insect groups
Ephemeroptera, Plecoptera, and Trichoptera (SEPT) can be used to assign a bio-
classification using DEM criteria. Data on the pollution tolerance of each taxa
has been incorporated into a biotic index which also can be used to assign bio-
classification.
Station descriptions are given in Table 5 and a diagram of the study area is
presented in Figure 3.
Station 03A - This site is located on UT Abernathy Creek, just above its
confluence with Abernathy Creek (near SR-1302, Gaston County). This portion of
the stream is large enough to have year-round flow, but portions of this wat-
ershed would be classified as intermittent streams. The extremely high conduc-
tivity (2600 umhos/cm) at this site probably reflects the input of chlorides from
the Lithium Corporation discharge, located about 0.5 miles upstream.
-14-

Stations 01 and 03 - These two stations were located on Abernathy Creek near
SR-1302 and 1-85, Gaston County: Station 01 was just above the UT and Station 03
was about 100 meters downstream. These sites correspond to the chemistry samp-
ling sites discussed in previous sections of this report. Station 01 was
slightly narrower than Station 03 and had a higher proportion of boulder/bedrock
substrate. Both sites, however, had good boulder/rubble riffles and would be
expected to have very similar macroinvertebrate communities. There was no visi-
ble evidence of the Lithium Corporation discharge, but conductivity increased
from 72 umhos/cm at Station 01 to 1350 umhos/cm at Station 03. Abernathy Creek
receives nonpoint source runoff from the Kings Mountain area.
Station 04 - This station was the toxicity test dilution water site at
Beason Creek (SR-2246, Cleveland County) . This stream is located in an agricul-
tural watershed and the substrate was mostly sand, reflecting severe erosion in
this area.
Stations U and R - The Biological Monitoring Group has previously (09 May
85) collected data from two other Gaston County streams as part of an investiga-
tion of the Gastonia WWTP. The upper part of Catawba Creek (above the WWTP
effluent discharge point) is listed as Station U. This site was effected by
urban runoff, and was very similar to Abernathy Creek in terms of size, sub-
strate, and land use. Data from South Crowders Creek, a relatively clean stream,
is included as a reference station (Station R) for this study.
Taxa richness values were used to assign a bioclassification for all sites
using DEM criteria for piedmont streams. Low taxa richness values (Tables 6 and
7) indicated Very Poor conditions at UT Abernathy Creek below the Lithium Corpo-
ration discharge (ST/SEpT = 25/0, Station 03A). The more pollution intolerant
"EPT" groups were completely absent at this site and the fauna was dominated by
pollution tolerant Siluliidae (blackflies) and Chironomidae (midges).
-15-

Taxa richness values indicated Fair water quality in Abernathy Creek above
its confluence with the UT (Station 01, ST/SEAT = 67/13), but Poor water quality
below the confluence (Station 03, ST/SEPT = 43/4), indicating degradation of
water quality due to the Lithium discharge. Thirteen EPT taxa were present at
the upstream site, including 7 abundant species. Both taxa richness and species
composition at Station 01 were very similar to data collected in 1985 at Station
U on Catawba Creek, another urban stream in Gaston County similar to Abernathy
Creek in terms of size, substrate, and land use. Only four EPT species were
collected at the downstream site on Abernathy Creek, and none were found to be
abundant. Like Station 03A, Station 03 was dominated by a mixture of tolerant
Simuliidae and Chironomidae.
The large difference between the Abernathy Creek sites was surprising in
view of the short distance (100 meters) between Stations 01 and 03. Several of
the EPT species at Station 01 are quite prone to daily downstream "drift", par-
ticularly Baetis flavistriga and Baetis 1p uto. Lfhe-.absence of-these potential
cozers at Station 03 clearly indicated that-the-discharge was-acutely-toxic
t'a these and other pollution-intolerant_stream-invertebrates. However, note the
higher taxa richness at this site as compared to Station 03A, suggesting a more
severe impact from the Lithium discharge on the UT as compared to this site.
High taxa richness was recorded at the toxicity test dilution water site
(Station 04, ST/SEPT = 69/17), and this station received a Good-Fair rating.
Most taxa at this site were confined to areas of woody debris or bank areas.
Differences between Station 04 and the reference site (Station R) probably are
caused by agricultural runoff upstream of Station 04.
Comparisons of biotic index values (Table 7) independently indicated the
same between-station patterns as had the taxa richness values. Biotic index
values gave a Good rating to Station R; a Fair rating to Stations U, 01 and 04;
and a Poor rating to stations 03A and 03. Both sets of data suggest that a water
-16-

Table 5. Station descriptions, UT Abernathy Creel; (Station 03A = BeIo'rr
effluent), Abernathq Creel: (Station 0i = Above UT, Station 03= Belo,,-,,, UT)
and Beason Creel; (Station 04 = Dilution ,,%.ater site), Gaston County, 10 ,dune
B7.
STAT I ON
1) 1 0 3A ��3 04
WIDTH (H) 4 2.5 5 4
DEPTH (P-1)
A�JERAGE 0. 7. 1
M AXII`11_M 0.15 0.5 0.0 �
C A N 0 P V 70 50 40 70
AUFWUCHS Moderate [laderate Abundant SIight
BANK EROSION Moderate Moderate Slight Severe
SUBSTRATE
BOULDER 60 10 10 -
RUBBLE "0 40 40 -
GRAVEL Trace �V z 5
S'1LT Trace Trace Trace 5
CONDUCT IV T'-r 72 2►D3_'1) 1350 -
WATER Clear Clear Clear Clear
-17-

quality ranking of Station R> Station 00 Station 01 = Station U> Station 03>
Station 03A. Data from Table 6 (taxa richness by group) was used to test for
significant differences between stations. A Wilcoxin signed-rank test (Table 8)
indicated that both Stations 03A and 03 were significantly different (5% level)
from Station 01. Furthermore, Station 03A was significantly different from Sta-
tion 03, again suggesting a more severe impact from the Lithium discharge on the
UT to Abernathy Creek than is seen on Abernathy Creek. There was no difference
between the control site (Station 01) and the dilution water site (Station 04)
indicated by the Wilcoxon signed-rank test.
Appendix A presents a list of all species collected in this study.
The Lithium-Corporation-effluent-was found to-cause-severe-Water_qualy
problems in both UT Abernathy`Creek andAbernathy-C er ek- It is not clear, how-
ever, how far downstream this effect may occur. High conductivity-values-be Tow
the ischarge_i.ndicated_that_Li.thium_Corporation adds large amounts of dissolved
solids-to-these_streams. TFe_-ef-fluent is_known_to�conta-in-high-concentrations_of
CH loride, although other dissolved materials-may contribute tb-the-observed_tox,-
i ity problems i7 Abernathy C� reeky The State of Kentucky has had extensive
experience with chloride problems, including both industrial dischargers and oil
drilling brine wastes. They have established a chloride standard of 600 mg/1,
equivalent to a specific conductivity of 1900 umhos. Kentucky dischargers have
attempted to resolve chloride problems by effluent removal, dilution, or waste
treatment (drying beds) .
CONCLUSIONS
On-site toxicity tests conducted on the effluent of the Lithium Corporation
resulted in a 48 hour Ceriodaphnia dubia LC50 of 21% (influent LC50 of 0.32%), a
96 hour fathead minnow LC50 of 70%, and a C. dubia 7-day LC50 of 11%. The
effluent characterization/fractionation tests conducted at the Aquatic Toxicol-
ogy Laboratory May 27, 1987 indicated that (chlorfde was a maja cr ontiibutor to
-18-

Table 6. Taxa richness, by group, UT Abernathy Creek (Station 03A),
Abernathy Creel: (Stations 01 and 03) and Beason Creel: (Station 04. Gaston
Count, 10 June 1987.
Station: ?1 03A 03 04 U 1 R
Group
EPHENEROPTERA 7 0 ? 10 11 17
PLECQPTERA 1 0 1) 2 _? 5
TRICHOPTERA S 0 1 5 3 9
COLEOPTERA 9 3 7 4 8
UDONATA 6 1 G 5
NEGALQPTERA 1 1 3
DIPTEI?A:MISC, 6 4 3> 3
DIPTERA:r_HIRON. 1 1 ' L.' 7 16 t_j
0LiG0CHAFTA 0 rl 5 4
CRUSTACEA 0 1 1 2 1
M 0LLUSC.A
OTHER 0 i 2
SUBTi iTAI 'FPT) 13 0 4 17 1 F, 71
,.
TOTAL r�7 `25 4.. Ll9 55 �to
RAT1NG Fair V. P„cjr Poor- Good-Fair Fair 61)lij'r
1Data from prior sarnplinq (09 Mau '35) in Ga-0cin County: "U" i an urban stream
��irrnilar to Abernathy Creel., Catawba Creek; at SR 2439; "R" is a reference site,
South Croor3-ders Creek at SR 1 103.
-19-

[!T A 3 Eti f - I.- f f`4-3. - rl 7 6 1.
JLllIll[101 Y JLOLI�LIUb, UI MUCI IIOLIiy L 1 tt:f, 4QL,]LIUII V,>M = UGIU'YY
effluent), Abernathy Creek (Station ()1= Above UT, Station 03 = Below UT)
and Beason Creek: ('Station 04 - Dilution water site'), Gaston County, 10 .June
B;.
S)TAT10NEi
i]1 03A 03 04 I i' R3
Total Taro Richness G;. -?S 43 b� JJ 89
EF'TTa,;aP,ic klnes s 1, Ca 17
1G 71
EPT Abundance 1 15 0 F, I l i C1, 13
Rating (EPT Taxa Richness) Fair V. Poor Poor- Good-Fair Fair Good
Biotic. 1ndex2
Rating (biotic index.) Fair PILL`r Poor- Fair Fair Ci ool'i
1 Abundant = 1 i_�, Common = -3, Rare = 1 : sur'nmed for tal l EPT tdxd
A4'erS1]W tolerance 'ti dWeS for a11 abundant ta xa
-Data from prior- sunplinq (C;ig May EJEJ) in Ga%--Jon n F-."ounty; ,.II" i are
°_.tr-earn similar- to Aberndthp Creek::, Catawba Creek- at SR 24'F-; "F" is
reference site, :_oath Crowders Cree'K at SR 1 1(-'13.
-20-

Vilcoxon signed--rank XI : sta 1 Y1 : sta 2
Number: s Rank:' Mean Rank:
- Rankf: 0 fr;.
+ Ranks 11
note 1 oases eliminated for difference= 0.:-:-. .:.
Z -2.934
Z con-ected for ties .-2•$44
tied groups 4
Vilcoxon signed-rank XI : sta 1 Y2: sta.3
Nun-,t-er-. 2 Rank.- Mean Rank:
- Ranks 2 7.5 3.75
+ Ranks y SS.S. 6.5
nm}te 1 cases eliminated fo.:difference = 0.
-2.6,
cc,rr-ected for ties
# tied qn ;s is
Yilcoxon signed-rank X1 : sta 1 Y3: sta 4
�Jurn�er : 2- Rank Wank::
- Ranks 5 26
+ RaFIL 15 I2g NS.-
nc,t. 2 c -1_e_ ellrninated fw- differe,io:, = n.
c*r-rec.tea ipr tie_ I -.1 rw {
tied groups 3
Table S . Statistical tests (Wilcoxon signed-rank ,test) of between-station
differences. Tests use taxa richness data (by group) , * = significantly
different at 5% level, NS = Not significant.
­21-

Table 8. Continued.
Wilcoxon signed-rank X I: sta 3 Y] : sta 2
Number: I Rank: Mean Rank:
- Ranks t 12.5 2.5
t Panks 8 42.5* 5.312
note 3 cases eliminated for difference = 0.
z -2.369
Z corrected for ties -2.399
* tipd groups 2
-22-

the recorded toxicity in that sample. Based on these toxicity results (on-site
and laboratory) and Lithium Corporation's instream waste concentration of 82.3%
during low stream flow conditions, it is predicted the effluent will be present
in the Um to Abernathy Creek at a concentration well above that level demon-
strated to be acutely toxic to C. dubia and Pimephales promelas (fathead min-
nows). The data also predicts that during periods of average receiving stream
flows (IWC=35.4%) the effluent will cause acute toxicity instream to organisms
with similar (and greater) sensitivity as C. dubia.
Analyses of_chemical_samples_show_that-of_the-metals detected-in fhe ef'flu-
ent oril'y Yithium was present iri coriceritrafions which could contribute_to_the
observed-to`xicity`ta'test organisms. El`evate`dl effluent-levels of-chloride-wer-e
detected howeve of in concentrations reported-to-have caused-acute-toxicEi y
in test organisiris. It is possible that`bacause ttie effluent-contained-both lith-
um-and ch'l'oride-that significant quantities of_lithium_chloride-were--present and
that-th s compound-was-contr-ibuting-to-the-toxic responses recorded-in-the
CCeraodaphn a-and-fathead minnow bioassays.
Chlorine, in the form of total residual chlorine (TRC), most likely contrib-
uted to the toxicity in all on-site bioassays conducted. TRC averaged 0. 7 ppm in
effluent used in the 7-day C. dubia chronic toxicity test, and was detected at
concentrations of 0.7 ppm and 147 ppm in the 48 hour acute C. dubia effluent and
influent tests, respectively. Residual chlorine was not implicated as a toxic
agent in the effluent sample shipped to the Aquatic Toxicology Laboratory for use
in the characterization tests. This sample contained 0.02 ppm TRC. The Oxidant
Reduction test was performed to determine to what extent oxidants, such as chlo-
rine, contribute to toxicity. Effluent toxicity was not significantly
altered after oxidants were reduced, possibly a result of the small quantity of
chlorine present in the sample.
-23-

No organic constituents were identified in the effluent and probably do not
represent a problem.
Analyses of benthic macroinvertebrate communities in the UT to Abernathy
Creek and in Abernathy Creek both upstream and downstream of its confluence with
the UT indicate that the effluent is seriously impacting macroinvertebrate biota
in both streams.
RECOMMENDATIONS
1. ) The Lithium Corporation should continue performance of the monthly
Pass/Fail Ceriodaphnia survival and reproduction tests at a test concentration
equal to 83% as stated in their NPDES permit #NC0005177.
2. ) Due to effluent levels of lithium, chloride, and residual chlorine
'efforts to reduce these constituents should be continued as outlined in the first
6-month report submitted to DEM February 4, 1988. Specific in-plant areas of
concern include reducing the quantity of lithium hypochlorite process wastewater
and metal washout wastewater discharged to the waste lagoon, thereby reducing
input of these toxic constituents.
`3. ) It is recommended that an effluent lithium monitoring requirement be
included in the subject permit at re-issuance. ,
-24-

FOOTNOTES
1 Hamilton, M.A. , R.C. Russo and R.V. Thurston. 1977. Trimmed Spearman-Karber
method for estimating median lethal concentrations in toxicity bioassays.
Environ. Sci. Technol. , 11(7):714-719.
2 Short-Term methods for estimating the chronic toxicity of effluents and
receiving waters to freshwater organisms. 1985. Eds. , W.B. Horning and C.I.
Weber. EPA 600/4-85/014.
3 Hansen, L.A. In Press. A method for chemical fractionation of chloride from
complex effluents.
4 Mount, D.I. and T.J. Norberg. 1984. A seven-day life-cycle cladoceran toxic-
ity test. Environ. Toxicol. Chem. , 3:433-442.
5 Benoit, D.A. and G.W. Holcombe. 1978. Toxic effects of zinc on fathead min-
nows Pimephales promelas in soft water. J. Fish Biol. , 13:701-708.
6 Anderson, B.G. 1948. The apparent thresholds of toxicity to Daphnia magna for
chlorides of various metals when added to Lake Erie water. Trans. Am. Fish.
Soc. 78:96-113.
7 Environmental Protection Agency. 1987. Sodium chloride round robin. Unpub-
lished results, EPA-Athens, GA.
8 Birge, W.J. , J.A. Black, A.G. Westerman, T.M. Short, S.B. Taylor, D.M. Bruser,
and E.D. Wallingford. 1985. Recommendations on numerical values for regulat-
ing iron and chloride concentrations for the purpose of protecting warmeater
species of aquatic life in the commonwealth of Kentucky. Memorandum of
Agreement No. 5429, Kentucky Natural Resources and Environmental Protection
Cabinet, Lexington, Kentucky, 70 pp.
9 Adelman, I.R. , L.L. Smith, Jr. , and G.D. Siesennop. 1976. Acute toxicity of
sodium chloride, pentachloro-phenol, guthion, and hexavalent chromium to
fathead minnows (Pimephales promelas) and goldfish (Carassius auratus).
10 Ward, R.W. , R.D. Griffin, G.M. DeGraeve and R.A. Stone. 1976. Disinfection
efficiency and residual toxicity of several wastewater disinfections. Vol. 1
- Grandville, Mich. EPA 600/2-76-156. 144 pp.
-25-

APPENDIX A. SPECIES LIST FOR LITHIUM CORP. STUDY
ASERNATHY, UT ABF_RNATHY AND BEASON CREEKS. R=RARE, C=COMM+ON, A=ABUNDANT
-------------------------------------------------------------------------
1 I STATION I
l --------------------------------------- I
i I I 03A I I I
! ! 01 --------- 1 03 1 I
i 1--------- 1 STREV! I ---------- 1 04
! I STRCA"i I --------- I STREAM I --------- I
I I --------- I UT I --------- I STREAti I
1 ! AP4ERNATHY I A P,ERNA T HY1 A3ERNATHY I —_-------
rR 1 CR fi CP. ( 01 1 E3EASON C.RI
--__------+---------+----------+--------- I
} { ---------+---------+---------+--------- I
I -------------------------------+---------+-----------+---------+--------- I
IORDER ISPECIFS f I 1 1 !
I ---------------t--------------- I ! 1 1
EPHEMER:`1PTERA 13AETI S ! 1 1 1 !
! !FLAVISTRIGA I A ! f I C !
1 I ---------------+----------+-----------+---------+--------- 1
i IBAETIS I I ! I
! 1INTERCALARIS I I 1 1 ' C
1 I ---------------+---------+-------------------+
1`sAETI S PLUTO I A I I I A I
1--------------- ----------+---------+---------+---------- I
I lJAETIS I I I I I
i I ?E?OPI`,�Q,IJUS ! R I I I A I
1 I ---------------.-------_---+---------+---------+--------- I
I I3PACHYCIERCUS I 1 1 I
I 1s?P I ' I 1 1 R 1
1 I ----------------+---------+----------«--------- --------- 1
1CAE,',.IS SP!' I I 1 1 a
l ---------------t---------#--------------------+---------' I
! !---------------_+------_-_+_--------+------------- ----------- I
} I 'll SPP I A 1 I R ! l
1 I-----------------+----------+---------+--------- t---------- I
1 IISJtfYCHI.A SPA' I A I I f A. I
1 I-----------------+---------+----------}---------+--------- 1
1 I P SEtiUDnCLOEDIN I I I 1 1
1SPP } I 1 1 I
! I----------------}---------+---------F----- !
1 1 STE"10"JE`Ea I I 1 I
IMCDESTW., I A ! I I A !
1-------------------------------------------------------
I ITRICJ! YTHODES I I I I
I ISPP I I 1 1 A I
I ---------------+---------------+-----------+-------------------+----------
I PLECOPT-RA IAMPHINEr"+URA SPPI I I I R I
-------------------------------------------------------------------------
(CONTINUEO)
—27—

APPENDIX A. SPECIES LIST FOR LITHIUM CORP. STUDY
ABERNATHY, UT ABERNATHY AND BEASON CREEKS. R=RARE, C=COM' ONO A-A9UNDANT
I I STATION 1
1 I ---------------------------------------- I
i I 1 03A I I 1
1 1 o I 1 ---------- 1 0.3 1 I
! I ---------- ! STREAM I--------- I 04 1
I I STREAM l --------- I STPEAt4 I-------•--- I
UT --------- I STREAM I
I A7 ERNATHY I A IERNATHY I AE•ERNATHY I ------•--___ I
I CR I CR I CR t0) l8r-ASON CRI
! I ---------+---------*---------+--------- I
! I I I II I
1 1 ---------------------+---------+--------- I
I 1 I I I I
I ----------------_-------_-------+----_----_+----------+------------------- I
109DER ISPECIES I I I I I
I ----------------+----------------- I I I I
IPLCCOPTERA IPERLESTA I I I I I
I IPLACIDA 1 R I I I A I
I ---------------+-----------------------•---+---------+----.-----+---------- I
I TR.ICHOPTERA iC".EU4ATrPSYCHF I I I !
1 ISPP I C I I I r, i
1 I---------------+---------+---------+----------t---------- 1
1 ICHIMARRA SPP I C I I I I
II----------------+---------+----------+ +---------- I
I IHYOR0PSYCHE I I I ! 1
I 13ETTENI I A I I I A I
II----------------+----------+----------+ --+---- I
I IHYDROOSYCHE I I 1 I 1
I I3PA;PNA I ' C 1 .1 i C I
1 I---------5-------+----------t-------------------+---------- I
•
! 1HYDROPTILA SPP I i I << I
! I ---------------+---------t---------+---------_- --------- I
! ILYPE DIVERSA I I I I P I
1----------------+---------+---------+---------+--------- I
I 1PYCN'lrlSYCIIE I I I I I
I IGUTTIEER I r I I I 1
II----------------}----------+---------+---------+--------- I
IPYCNO°SYCH EI I I I f
I ILEPIDA
--------------- ---------------}---------+---------+---------+--------- I
ICOLEOPTERA ID Rnslis SPP I C 1 C I C I I
II +--- ---+---------}----------+---------- I
I lCgPELATUS SPP I C I I . C I I
II---------------+----------r---------+---------+----------- I
I 10INFUTES SPP I C I A I A I A I
1---------------r---------+------------------------------ I
I I ENOCH?.US SPP I I I r I R. I
! I ----+---------+---------+---------+--------- I
! lHF-LICHUS SPP I R I I R I c
--------------------------------------------------------------------------
[CONTINUED)
-28-

APPENDIX A. SPECIES LIST FOR LITHIUM, CORP. STUDY TUESDAY ,
A3ERNATHY9 UT ABERNATHY AND BE.ASON CREEKS. R=RARE9 C=COt4" ONq A=A4UNDANT
--------------------------------------------------------------------------
1 1 STATION
1 I ---------------------------------------- I
} 1 I 03A I I !
} 1 n i I --------- 1 03 1
1 1--------- I STREAM I--------- 1 04 1
i 1 STREAM I --------- I STREAM { -------
! -- ------- I UT I--------- 1 STREAM I
I I A BE-RN ATHY I A DEP NATIIY I ABFRNATIIY I ----------- I
I I C z I cF I cR ( n ) 1 ac-ASONI CR I
I ---------+---------+----------*--------- I
1 1 I I V I
} I ---------+----------+----------f--------- I
I ! 1 I 1 1
} ------_-------------------------+-------- +- --+---------+-------_ I
I OR')FR I SPEC I=S 1 I I I I
I ------------------+---------------- I I I I I
iCnLFOPTERA IHYDROCHl.1S SPP I I I R I I
II-----------------+---------+---------+---------+----------- I
I IHYDROPORUS SPP I C I I I R I
II ---------------+---------+---------}---------r---------- I
I ILACC` PHTLUS SOPI R I R 1 I I
I1 ----------------+-------------------+---------*--------- I
I 1AACRONYCHUS I I 1 I 1
! IGLABRATUS 1 I I I R I
II -----------------+---------+---------+---------+--------- I
1 WELTODYTE S SPP ! '? 1 I € 1
II ----------------{---------+---------+----------+--------- I
1 IPROYORESIA 1 I 1 1 I
} I ELEGA".AS
II ---------;7------+----------}---------+---------------_---- I
I 1 SPEQCHOPSIS € I 1 ( 1
I ITFSSELLATIJS I P
} I----------------}----------+---------+--------- +--------- I
1 ITROPISTERNUS I I I I 1
1 ISPP I R I I I �' 1
1 __ ---7----------+-----------------+---------+---------+---------*--------- I
IODONATA 1ARGIA SPP I A 1 A. I A I !
II ---------------+---------+---------4------ t---------- I
} 12,OYERTA VINOSA I R I I I C I
1 I---------------+_--------+----------+---------+--------- 1
} ICAL9PTERYX SPP I C i € 1 c I
iI--------------- ---------+---------}---------+---------- I
I IC0R7'ULFGASTFR ! 1 I I I
I ISPP I I I I R I
II ---------------+---------F---------+----------+--------- I
IGOMPHIAS SPP I C I I 1 C I
I---------------+--------- ----------+----------+---------- !
I 1HAGFNIUS I I I I I
1 1 =3REVI STYLUS I 1 I I P.
-------------------------------------------------------------------------
(C?NTI NVF--0)
-29-

APPENDIX A. SPECIES LIST FOR LITHIUM CORP. STUDY
ABERNATHY, UT ABERNIATHY AND f- EASON CREEKS. R=RARE, C-COV.FiONV A=ABUNDANT
-------------------------------------------------------------------------
1 1 STAT ION I
II --------------------------------------- 1
! I I 03A I i I
! 1 0 L I --------- 1 0 3 1 I
1 I --------- 1 STREAM I --------- 1 04
I 1 STREAM I --------- I S TRI`AM I --_------- I
1 1 --------- I UT I --------- I STREAM 1
I 1 ADERNATHY I ABER.NATHY I .A`3cRNRT;IY I --------- I
I I CR. I CR I CR ID) IBLASON CRI
II ----------+---------r---------+--------- 1
I I I I V I
1 I ----------+---------+---------+--------- I
I ------------------------------ }---------+----------}---------+---------- I
IORDER I SPECIE S
I ----------------+--------------- I I I I 1
ODONATA I LA;NTHU S
I IPARVULUS ! 1 I I P
II---------------+---------+--------- ---------+---------- I
I ILESTE.S SPP I R I P. I I
II ---------------+---------+---------+---------+---------- I
! I OPN74c,Op-PiquS I 1 I I I
1 ISQP I R I I I R I
I ----------------+---------------+---------+----------+---------+----------- I
IMC-GALOPTERA 1GORYDALUS I I I I 1
I I C"P,AI U T U S I C 1 R. 1 R I R. I
II -------------------------------------------------------
I I S I.ALIS SPP I A I I C I c !
I ----_------------+---------------+---------+----------t-------- }- i
IDIPTERA: CHIROM ! AL'L45r-S ,Yjs I I I 1
f I MALLOCHI I C I C. I A I C
II ----------------+---------+----------t---------+--------- I
i t5 RILLiA SPP I I I I �. 1
II ---------------+--------------------+---------F--------- 1
i lCAP;�If"?CLAI�IL�s 1 I i I I
1 ISPP I A I C 1 A I R I
II ---------------+--------------------+----------+--------- I
I ICHIRCIINJ,.US SPp 1 A I C. I .A I C. I
II ---------------+---------+---------}----------+--------- I
! I CONCHAPFLUP IA I I I I 1
I I GRIOUP I A I A. I A I A
I ---------------+---------+---------+----------+--------- I
I 1 CORY`•ONFUR.A SPP I P, I I I R
I -----------------+------- +- -}---------}--------- I
I ICRICOTOPUS/ORT-1 1 I ! !
I IHOCLADIUS SP1 I
I---------------+---------+---------+----- *----- I
ICP.IC0T0PUS/0R7- 1 I I I !
1 1tIOCLADIUS SP3 I A I I A I I
--------------------------------------------------------------------------
(CONTINUED)
—30—

APPENDIX A. SPECIES LIST FOR LITHIUM CORP. STUDY
A3ERNATHY, UT ABERNATHY AND BEASON CREEKS. R=RARE, C-COMMONT A=AAUNDANT
-------------------------------------------------------------------------
i I STATION 1
iI -_------------------------------------- I
I 1 03A I l 1
I 1 01 f --------- 1 03 ! I
1 I ---------- f STREAM I --------- 1 04 1
1 1 STREAM I --------- 1 STREAM I --------- I
---------
--------- UT I --------- I STREAM !
1 IABERNATHYIABERNATHYIA�3ERNATHYI--------- I
I CR 1 CR I Cfi ( D} IDEASON CRI
-------- ----------+-----------f---------- I
1 1 I I I' I
II ---------+---------+---------+--------- I
1 1 , I I I !
I -------------------------------+---------+----------+------------------ I
1 ORDER I S('ECIES I I I I !
I _---_-___.----_-+--------------- I I ! ! 1
IDIPTERA:CHIRON 1CRICOTOPUS/ORT- 1 I ! I I
I P10CLAOIUS SP5 I A I A I A f 1
fI ----------------+---------+--------- ----------+---------- I
I ICRTC0T0PUS/0RT- 1 I I I 1
1 IHCCLA^IUS SP6 I P. I C I I 1
II ---------------+----------+---------+---------}--------- I
ICRYPTOCHIRONO"A- 1 I I I I
IUS FAVUS I C I I I C I
II--------- ------+-_--------+-----_---+---------+--------- !
I IDIAME-SA SPP I C I R. I A I 1
II ----------------+---------+---------+---------+---------- I
I JC)TCROTE�' DIPES I I 1 1 !
1 IspP [ ' I A I A I 1
II -------- ------+-------_-+---------+---------+---------- I
EUKIE�:EERIELLA I I I I I
I 1Sp1 I I I I R 1
II -------------------------------------------------------
I EUKT r-ZF-f-ER I ELLA I 1 I I 1
I ISP11 I r I I A I I
1 I --+----_--_--+---------+---------t----------I
1 IMICROpSECTRA I I I I I
I ISF5A I I R I R I I
II ---------------+---------+---------+---------*--------- I
I IMICROTE`JOIPFS I I I I f
I Ispl 1 R I I I I I
11 ---------------+----------+----------+------------------- I
! INATARSIA 5PP I C I c I A 1 rl
1 1
II---------------+---------.---------------------+--------- 1
f I PARACLAPOPELMA I I I I I
I I SIPP I I I I R I
----------------+---------+---------+----------+---------I
I IPARAPHAENOCLAO- 1 I I I 1
I WIS SPECIES 1 I 1 I I R 1
-------------------------------------------------------------------------
( CONTINUED)
-31-

APPENDIX A. SPECIES LIST FOR LITH.IU`4 CORP. STUDY
ASERNATHY9 �JT ABERNATHY ,AND REASON CREEKS. R=RARE, C=COMr10,N• A=ADUNDANT
-------------------------------------------------------------------------
I STATION I
II ------------------------------------------ I
! I I 03A I I t
! 1 ni 1 --------- 1 03 1 t
I I ----- ---- i STREAM I---------- 1 04 !
I I STREAM ! --------- I STREAM I --------- I
I I --------- I UT I --------- I STREAM I
! lASERNATHYIABERINATHYIABERNATHYI --------- I
I I CR I CR I CR ( 0) IBEASON CRI
! I ---------.-----------+----------+--------- I
I I I I I �
! I--W--------+--------------------+--------- I
I I . I I I I
I -------------------------------+---------+---------t----------+--------- I
IORDE-R. ISPECIES ! ! I 1 !
I ---------------+---------------I ! I f 1
10IPTER.A: CHIRDN I PAR ATE`1OIPES I 1 I k I
! I SPP I I A I
II---------------+---------}----------F---------•--------- I
I IPHAENOPSECTRA I I I I I
I 1FLAVI?ES I I C I p I i
iI-------------------------------------------------------
PH A EN'9P SECTRA 1 f I 1 I
! I SPA
---------------+---------+-------- +---- --+-----
1 1 POLYPEDTLW-1 1 I I !
I IAVIrF-PS
1 ----------------*---------+---------+------
! I PCLYP�:DILU ', 1 I I 1 !
1 iCOPd�/LCTuf;. ! ! I 1 A
1 1 -------+--------- +---------+----------+--------- I
I IP 0 L Y?E Di ILU'A I I I I I
! IFALLAX I I I I R I
II ---------------}----------+---------+---------t---------- I
1 I POLY?Ef�ILIJ`+ 1 k I I !
I IHALTERALE I R I f I I
iI---------------+-------_-_+---------}---------}--------- 1
I IPOLYPEOILOM I I I 1 I
I IILLIN7E^:SE I A I t I A I C
! I ---------------}-------------------+----------}--------- 1
I IP0LYPEOILW1, I 1 1 1 I
1 ISCALAF I I R I c I f !
! I ---------------+---------+--------------------+--------- 1
1 IPROCLACTUS SP^ I A I C I A I f I
II-----------------+---------+----------t----------+---------- I
i IRHEOCRICOTOPUS I I I I I
I Ispi I I I I c I
II-----------------+------------------------------+---------I
I IRHEOTAMYTARSUS I I I I I
I IsPP I I I ! !
-------------------------------------------------------------------------
( CONTINUED]
-32-

APPENDIX A. SPECIES LIST FOR LITHIUM CORP. STUDY
ABERNATHY, UT A.BERNATHY AND 'SEASON CREEKS. R=RA€?E9 C=COMi ONO A=ABUNDANT
-------------------------------------------------------------------------
! 1 STATIO^f 1
II -------------_----------------------
i 1 I 03A I 1 I
I I Di I -- ----- 1 33 1 I
I f ---------- I STREAM I ---------- 1 04 1
STREAM I ---------- ? STREAM I ---------
--------- I UT I --------- I STREAM
I I AI?,ERNAT1AY I A,3ERNATHY I A�3EPNATHY I ----------
I I CR I CR I CR ( D ) IBEASON CRI
? I ---------+---------}----------+--------- I
! I 1 I ! ' I
II ------_----+--------------------+---------
I I I I I I
I -------------------------------+---------+--------- +----------}--------- I
IO2DER ISPECIES ! I I I 1
i---------------+-----_--------- I 1 I I I
IDIPTERA:CHIRON IROBACKIA I 1 I I I
! IDEMFIJEREI I ! I I A
! I -----------------+----------+-----------------------+--------- I
I ISAETHERIA TYLUSI I I I C I
II----------------*----------+---------+---------t--------- I
! I STEMOCH IRO-N- NUS I I I I I
! 1 SPP f ( I I I
! I ---------------+---------+---------+----------+--------- I
ITANYTARSUS sPz 1 A I A I A I C i
! I------------------+----------+----------+---------+----------
I 1THIr='lE?aA��IELL4 I I I I I
II -------_---------+-------------------+----------+--------- I
1 ITRIE;'=L )S iSPP I C I I P I A
---------------+----------------f---------+---------+----------+---------- I
IDTPTI PA: MISC 1ANOPH LFS SPP I ?. I I I f
I1 -----------------*---------+----------+---------+--------- I
I !L# PIDID_' C
I1 ----------------+---------+----------+------------------- I
i iPALI• O!lYTA I I I I I
I I ( C;)"PLEX ) I R I C I C I P I
Ii-----------------+-------------------+---------+---------
I I S T MUL I UM I I I I 1
1 1VFINUSTUM I A I I I !
II ---------------+----_----+--------------------+--------- 1
I I SIMULIU�; I I I I !
IVITTATUM I A I �L I A I C 1
II ----------------+------_--+---------+-----------------
1 ITIPULA SPP I C I I R I A !
I - t---------------r-----_---+---------+---------+--------- 1
IOLIGOCHAETA 1AIJLODRiLUS I I 1 1
1 1LTrlN-3°-IUS I Q. I I I !
---------------------------------------------------------------------------
( CONTINUED)
—33—

APPENDIX A. SPECIES LIST FOR LITHIUM CORP. STUDY
ABERNATHY, UT ABERNATHY AND PEASON CREEKS. R=RARE, C=COMMONg A=A;?UNDANT
-------------------------------------------------------------------------
1 1 STATION I
I ---------------------------------------- I
I I I 03A I I I
I OI 1 --------- 1 03 1 I
1 1---------- I STEAM I --------- 1 G4
! 1 STREAM _ --------- I STREAM I --------- I
I I --------- 1 UT I ---------- I STREAM
I I ADERNATI',Y I A RERNA TF-,Y I AIIE RNATHY I ---------
I I CR I CR I CR ( 0) 1BEASON CRI
4 I ---------+---------*----------+--------- I
1 I I I I� I
iI ---------+---------+---------+--------- I
I I I I I I
I -------------------------------+---------+---------+---------+---------- I
ORDER ISrFCIFS I I I I I
1 ---------------+---------------- I
IOLIGDCHAETA IILYODRILUS I I I 1 1
ITEMPLFTgNI 1 I 1 ! R I
II--------------------------+---------+----------+--------- I
i lHUr FMCISTFRT I C I 1 I C I
1 I--------------- }----------+----------+----- +--------- I
! lLUM�,R'vULTDAE 1 A I I I C I
i1 ----------------- F---------+----------+----------+---------
f I 1AI< SI]p 1 C I I I R 1
! I ----_------_----._--------+--------------------+--------- I
i IOPISTHOPORA SPIT P, I I I C I
lI---------------}---------+---------+---------+---------- I
I ISLAVINA I ' I I I I
I IAPPFNOICULAT A I R I I I I
i --------------- +----------------+---------+----------+---------+--------- I
I CRUSTA.CEA I CA` FI ARUS SPA' I I I R I 1
I ---------------------------------+--------- +---------+---------+--------- I
IMOLLUSCA IPHYSELLA SPP I I I ! I
II ---------------+---------F---------+---------+----------- !
1SI'HAFRIUM SPP I C I I I I
I ---------------+---------------+---------+-----------+--------- }------- -- 1
IQTHER 1PROST+OMA I I I I I
IGRAECc"NS I R. I I I I
II----------------+---------+----------}---------- --------- I
I I SIGARa SPP I C I I R i
-------------------------------------------------------------------------
-34—

48 Hour Cladoceran Screening Toxicity Test Appendix
Aquatic Toxicology Group
N. C. Division of Environmental Management
The Aquatic Toxicology Group performs 48 hour static toxicity tests using cladocerans Daphnia pujgx and/or
Ceriodanhnia dub1a to estimate the toxicity of waste discharge to aquatic life in receiving streams. All lest and
sampling glassware and equipment with are reused are washed with soap and hot water, then rinsed in nitric acid,
acetone, and distilled/deionized water, to remove toxins and contaminants. Effluent samples are collected by DEM
Regional Office or Aquatic Toxicology personnel. All samples are collected chilled and below chlorination unless
otherwise specified. Each sample is collected as 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 within 72
hours after collection.
The effuent samples are prepared for testing by being thoroughly mixed, allowed to reach standard test
temperature, and aerated if dissolved oxygen is below 407 saturation. Total residual chlorine is measured. The
effluent is then diluted with culture water, typically to seven concentrations (with replicates)from 0 to 90%
effluent Each test chamber receives 100 mis total volume and ten test orgaoisms,0-24 hours old. Initial DO and pH
are measured in separate surrogate vessels of dilution and effluent solutions. The Lest is conducted in a 20 degree
centigrade incubator with a 15:8 hour light:dark cycle. Mortality of the test organisms is recorded after 48 hours,
along with final pH, dissolved oxygen,and temperature.
A 48 hour LC50, or concentration of effluent fethal to 509. of the test organisms in 4B hours, is calculated from
the mortality data using Trimmed Spearman-Karber.anolysis. An instream waste concentration (IWC) for the
effluent in the receiving stream is calculated using the wastewater treatment system permitted flow and receiving
stream 7Q10 now. The LC50 and IWC 'are then used to predict instream toxicity.
Guidance Documents:
1955. U. S. E. P. A. Methods for measuring the acute toxicity of effluents to freshwater and marine organisms.
Third Ed. (EPA/600/4-85/013).
1977. Hamilton, [I. A., Russo, R. C.. and Thurston, R. V. Trimmed Spearman-Karber Method for Estlmatinq Median
Lethal Concentrations in Toxicity Bioassays. EnyironmenLQI SdTechnology. Volume 11, Number 7, July
1977.
—35—

96 Hour On-site Toxicity Evaluation Appendix
Aquatic Toxicology Group
N. C. Division or Environmental Management
For each on-site toxicity examination, a pre-test inspection of the facility site is performed in order to:
l)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 and equipment needed for such. Determine discharge schedule.
Sampling is done below chlorination unless otherwise specifled.
4)Determine possible areas for dilution water collection (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 acute and static renewal
CeriosLaphnia a&a reproduction toxicity tests to determine the range of concentrations of effluent to be used for
the flowthrough toxicity test, to Lest 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 Lest procedures and requirements with appropriate facility personnel.
6)Determine appropriate sampling sites and techniques for benthic macroinvertebrate surveys.
All Lest and sampling glassware and equipment are washed prior to use with soap and hot water, then rinsed in
nitric acid, acetone, and distilled/deionized water to remove all toxins and contaminants. 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 (containinq approximately
one liter of dilution water), Len fish to a chamber.This transfer is accomplished five fish at a Lime 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 flowthrough toxicity test 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 7 day �Nrioda h;& static renewal reproduction tonicity test using nev,born organisms is begun Lhe first
day on--site. The organisms are transferred tc `resh 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; _imuiative
reproduction is calculated for each concentration. The test is conducted at 25 degrees centigrade with a 16 light:8
dark hour photoperiod. Test organisms are fed 0.1 ml of a yeast/alfalfa/fermented trout chow mixture with
Selenastrum canricornutum added per organism per day.(See Ceriodaphnia dubia Reproduction Toxicity Test
Appendix,)
Individual chemical/physical parameter meters are calibrated daily according to DEN standards. At 15 minute
intervals throughout the test, Hydrolab systems measure and record dissolved oxygen, pH, temperature, and
specific conductance in the Lest chambers with the highest and lowest concentration of effluent. These systems are
calibrated at test initiation, mid-point, and termination. Data from these systems is recovered daily and stored on
floppy disc and hard copy.Daily residual chlorine measurements will be made of effluent, influent, dilution water,
and receiving stream samples as feasible.
During the on-site evaluation, Biological Monitoring Group personnel collect benthic macro'snverLebrate samples at
the upstream, downstream, and dilution sites (see Benthic MacroinverLebrate Survey appendix,).Where
appropriate, electrofishing is undertaken upstream and downstream of the discharge to obtain resident fish
population data. On a site-specific basis,various other efforts are undertaken, such as moniLorinq_ dissolved
oxygen levels in Lhe receiving stream.
On a daily basis, Lest 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 alternate days, depending on need. 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.
—36—

Two separate 24 hour composite samples of effluent are collected for chemical analysis by means of an automatic
sampler. lnfluent, receivinq stream, and dilution water samples are also taken for chemical Lesling.
Static 48 hour cladoceran toxicity tests are conducted on a 24 hour composite sample of the effluent and a grab
sample of the influent.
A Lour of the facility is conducted. The actual treatment process is reviewed to ascertain the quality of the
operation of the treatment system and to determine the treatment system's appropriateness to Lhe type of waste
being treated. An inventory of any industrial contributors to a municipal waste treatment facility is made. The
manufacturinq process at on industrial facility is reviewed to determine the nature and composition of the waste,
An inventory of all chemicals used at the facility in manufacturing or wastewater treatment is made.Where
feasible, 48 hour cladoceran static toxicity tests may be performed on samples from individual wastewater
streams coming into the wastewater treatment facility to attempt to pinpoirFL a particular source of toxicity.
A photographic record is made of the manufacturing and treatment facility, sampling points, receiving stream, and
sampling procedures.
At 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. The Ceriodaohnia dubia reproduction toxicity test is continued al the lab until the 7th test
day.
—37—

Qeriod hnia dubia Reproduction Toxicity Test Appendix
Aquatic Toxicology Group
N, C. Division of Environmental Management
The cladoceran CerlodaQhnia dubia is used as test organism in a 7 day static renewal toxicity Lest. This test
estimates Lhe effect of an effluent or other water sample on reproduction. A control and 8 concentrations of
effluent ranging from 0.017 to 1007.. are used. There are 10 organisms per concentration, each organism in a one
ounce polystyrene Lest chamber with 15 mis of solution. The test is conducted at 25 degrees centigrade with a 16
light/ 8 dark hour pholoperiod.
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 and contaminants. Effluent samples are collected by
DEM Regional Office or Aquatic Toxicology personnel. All samples are collected chilled and below chlorination
unless otherwise specified. Each sample is collected as 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 within
72 hours after collection,
The effluent samples are prepared for testing by being thoroughly mixed, adjusted to standard lest temperature,
and aerated if dissolved oxygen is below 5 mg/l, Total residual chlorine is measured.
The test is initiated with organisms less than 24 hours old and within 4 hours of each other. The test is begun when
the neonates are introduced into the test chambers. Temperatures must be within I degree centigrade ror transfer.
The organisms are transferred daily to new test chambers containing freshly mixed solutions. Dissolved oxygen,
pH, and temperature are measured twice for each batch of test solutions. The initial value is Laken before the
organism is introduced and the final value after the organism has been transferred out the next day. The organisms
are red daily.Each organism receives 0.1 ml of rermented trout chow-yeast-alfalfa food with Selenaslrum
capricornutum added. t.
As reproduction begins, only the original Lest organism, now an adult, is transferred to the new chamber. A drop
of concentrated nitric acid is added to the old chamber. This kills the young so they can be easily counted under a
dissecting microscope.A mean number of young produced per adult is calculated for each concenLration. Mortality
of greater Lhan 20: in control Lest organisms invalidates a test.
Guidance Document:1935. U. S. E. P. A. Methods for estimating the chronic Loxicity of effluents and receiving
waters to freshwater organisms. (EPA-600/4-85-014)
-38-

5
Benthic Macroinvertebrate Sampling Procedure Appendix
Biological NoniLoring Group
N. C. Division of Environmental f1anagement
Benthic macroinvertebrates, found on the bottom of streams, rivers, and lakes, are commonly used as
biological indicators of water quality. The Biological Monitoring Group uses a standardized qualitative
collection method designed to sample all habitats within a wadable stream and provide a reliable
estimate of both the number of different kinds of organisms (taxa)present and their relative abundance.
This data is then used to assign water quality ratings to the stream and river. This methodology is
applicable for most between-site and/or between date comparisons.
The sampling methodology requires that freshwater streams or rivers be wadable. High water
conditions severely impair sampling efficiency by making critical habitats inaccessible. Ten samples are
collected and processed at each site: two kick net samples from riffle and/or snag habitats, three
sweep net samples from bank, macrophyte, and root habitats; three fine-mesh samples from, rocks,
logs, and sand; one leaf pack sample collected in the current; and a visual inspection of large rocks and
logs. A collection card is filled out at each sampling station with relevant data on station location, field
parameters, instream habitat, and water chemistry.
Data output for the standard qualitative technique consists of a list of all Laxa collected with a rough
estimate of abundance (Rare if 1-2 individuals are collected, Common for 3-9 individuals, or Abundant
for more than 9 individuals). The total number of Laxa collected or total Laxa richness (ST) and Laxa
richness for the pollution intolerant groups Ephemeroptera. Plecoptera, and Trichoptera (SEPT) are
calculated for each sample. These values ore used to assign a biological classification to each station
(Excellent, Good, Good/Fair, fair, and Poor). Bioclassification criteria for several ecoregions have beep.
developed, including mountain, piedmont, inner coastal, and outer coastal. The "bioclassification" rating
primarily reflects the influence of chemical pollutants. The effects of sediments are poorly assessed by
Laxa richness analysis. ,
An abbreviated version of this qualitative collection technique, the "EPT" survey, can be. used to quickly
determine gross between-site differences in water quality. Collections focus on the pollution intolerant
groups within the benLhic community: Ephemeroptera, Plecoptera, and Trichoptera. Only four samples
are processed: 1 kick, 1 sweep, 1 leaf--pack, and I visual. Field notes record extremely abundant Laxa.
Data summary is usually limited to EPT Laxa richness (SEPT) and EPT abundance (NEPT). Abundance
values are calculated using 1 for Rare species, 3 for Common species, and 10 for Abundant species.
These values are then summarized for all EPT Laxa.
—39—

List of Definitions
Aquatic .toxicology Group
N. C. Division of Env€ronmonLal Managoment
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 on 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 Guality Section) which performs
aquatic toxicity tests for the Division of EnOronmental rlanagement. The Group is located at the Cary
laboratory facilities. All test organisms (including Daphnia up lex, Ceriodaphnia ss., and fathead minnows) are
cultured at these facilities by Aquatic Toxicology personnel.
Benthos/Benthic ►nacroinvertebraLes - a wide assemblage of invertebrate animals (insects, crustaceans, molluscs,
etc.)which live in streams, are on important food source for fish populations, and are used as long terra water
quality indicators.
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 dubia -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 adapted for aquatic toxicity 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" toxicity testing for mortality, time
to sexual maturity, and reproductive rate. Ceriodaphnia dubia 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 (e.g. reduction in growth,
egg production, predator avoidance, feeding rate, etc.). Tests for chronic toxicity are frequently performed
during the entire life cycle of the organism,
Chronic value (ChV) - A numeric value representing the geometric mean of the numeric values of concentrations
analyzed as the No Oberser4d Effect Concentration (N.O.E.C.) and the lowest Oberserved Effect Concentration
(L.O.E.C.) by chronic toxicity testing. The chronic value is an estimate of Lhe toxicant concentration that will
be the actual no effect concentration based on the chronic effect tested.ChV=Antilog((Log 10L.O.E.C.+
Log 10€ ,O.E.C.)/2)
Cladoceran -Commonly known as water fleas, the Order Cladocera belongs to the Class Crustacea which includes
shrimps and crabs. Cladocerans are capable of asexual reproduction and therefore create genetically similar
offspring easily cultured in the laboratory environment,making them ideal as test organisms. The cladocerans
are generally considered to be a freshwater species sensitive to the effects of toxicants.
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 Lime at the some
location. This is frequently accomplished with automatic sampling devices.
Daphnia up iex -a small cladoceran crustacean. It is found throughout most of North America and obtains a maximum
size of approximately 3.5 mm. This organism has been adopted for aquatic toxicity 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 46 hour static toxicity testing for mortality. D.aqlex is widely accepted
in the field of aquatic toxicology for testing in moderately soft waters.
Design flow (DO -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 (rngd).
Dilution(water) -the water used in aquatic toxicity 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).
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1 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 or
pre-Lreated waste to some other facility.
FaLhe-.d minnow (Pimephelas rop molas) -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 cultures for use in toxicity testing_The fish can
produce eggs year round in the laboratory environment under correct conditions, to provide Lest organisms as
needed.
Flow-Lhrough - the flow-through toxicity test utilizes a mechanical dilutor which either continuously or occasionally
replaces the effluent/toxicant and dilution water in the test chambers with fresh solutions throughout the test
in an attempt to sample the variability of effluent toxicity throughout the test period.
Hydrolabl� -a multiparameter instrument which measures and records temperature, pH, dissolved oxygen,and specific
conductance of water. '
Insiream waste concentration(IWC) -the percent concentration of an effluent/Loxicont which is present in a stream
under worst case conditions (defined as 7Q10 tow flow). The IWC is derived from the formula: [PF / (7010 +
PF)) x 100 = IWC (%), where PF is the permitted flow (in cfs)of the facility in question and 7Q10 is the 10
year, 7 day, low flow (in cfs) of the receiving stream.
LC50 - that concentration or percentage of a waste/chemical/substance which is lethal to 50% of test organisms over
a stated period of time.
Lowest Observed Effect Concentration (L.O.E.C.) -The lowest concentration of toxicant to which organisms are exposed
in a life-cycle or partial life-cycle test, which causes a statistically significant adverse effect on the
observed parameters (usually survival, growth, reproduction ,and/or egg hatchability),
NPDES - National Pollutant Discharge Elimination System. A system devised by the Federal Government and adopted by
North Carolina for the permitting, monitoring, and pollution abatement of dischargers to surface v�aLers.
Neonate -roughly translated to newly born. In reference to cladoceran, the neonate refers Lo the life stage in the first
and early second instar, generally the first 24 hours of its life.
No Observed Effect Concentration (N.O.E.C.) -The highest concentration of toxicant to which organisms are exposed in
a life-cycle or partial life-cycle test, which causes no statistically significant adverse effect on the observed
parameters (usually survival, growth, reproduction ,and/or egg hatchability).
Permitted flow (PO -the volume of water and waste that is allowed by the NPDES permit to pass through a facility or
waste treatment plant. Permitted flow is usually expressed in millions ofgallons per day (mgd).
Screening toxicity test - a testing system established to determine general levels of acute toxicity of
compounds/discharges using short-term toxicity LesLs with sensitive species.
7Q10 - the measurement of a sLream's lowest average daily flow over a 7 day period during a 10 year spar, generally
stated as flow in cubic feet per second (cfs).
Sodium lauryl sulfate (SLS) -a chemical accepted by EPA as 3 toxicant for quality assurance testing of the health of
aquatic organisms.
Static - refers to ar. aquatic toxicity Lest in which toxicant/effluent concentrations are set up at the beginning of the
test and not changed or replaced 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, 0, e. genus, order, species).
Taxi richness -number of taxa.
Toxicity - the adverse effect of a chemical/substance on an organism, Toxicity is usually defined as a fatal or
non-fatal response over a given period of time.
Toxicity Test -a test used to determine the effects of a chemical or substance on an organism,
1JT or Unnamed tributary -a term given to streams which have no accepted name.
"Use or this term or system does not constitute an endorsement
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