sign in sign up
<<
Home , Mercury in fish

I ' U.8. T1SH &WIUMJFE SERVICE

PC-88-4-118 FLORIDA

I AND CONCENTRATIONS IN OF THE

ST. VINCENT NATIONAL WILDLIFE REFUGE PUBLICATION No. PCFO-EC 94-08

U.S. AND WILDLIFE SERVICE ECOLOGICAL SERVICES PANAMA CITY FIELD OFFICE 1612 JUNE AVENUE PANAMA CITY, FLORIDA 32405

Michael S. Brim Environmental Contaminants Specialist

Diane H. Bateman Assistant Environmental Contaminants Specialist

Robert B. Jarvis I Biological Aid

Gail A. Carmody Project Leader

1994

U.S. FISH AND WILDLIFE SERVICE / SOUTHEAST REGION / ATLANTA, GEORGIA

I . memorandum DATE: April 12, 19;94 REPLY TO Refuge Manager, FWS, St. Vincent NWR

SUBJECT: Mercury Report

TO: -Michael- S. Brim, Environmental. Contaminants Specialist,' ES,' FWS, Panama -.City, Florida ' .

I. have reviewed: the report entitled Mercury and' Selenium Concentrations in Fishes of the St. Vincent National Wildlife Refuge^ I concur with the repor't and particularly the "Site De's-cription" and "Conclusions and Recommendations11 sections. Thanks for the opportunity.

Qce_

osin

OPTIONAL ^O,RM^^ NO.. 10 (REV. t-80) GSAFP;I$IR C

o U.S. GPO: l

ST. VINCENT NATIONAL WILDLIFE REFUGE

• ^ . . . a •

PUBLICATION NO. PCFO-EC 94-08

U.S. FISH AND WILDLIFE SERVICE ECOLOGICAL SERVICES PANAMA CITY FIELD OFFICE ' 1612 JUNE AVENUE PANAMA CITY, FLORIDA 32405 (904) 769-0552

MICHAEL S. BRIM ENVIRONMENTAL CONTAMINANTS SPECIALIST

DIANE H. BATEMAN ASSISTANT ENVIRONMENTAL CONTAMINANTS SPECIALIST

ROBERT B. JARVIS BIOLOGICAL AIDE

GAIL A. CARMODY, PROJECT LEADERr

1994 TITLE: Mercury and Selenium Concentrations u* Fishes of the St. Vincent National Wildlife Refuge

ABSTRACT: From February 16 to May 6, 1988, sixteen lafgemouth (Micrdpterus salmoides), fifteen bluegill (Lepomis macrochirus), and thirteen brown bullhead (Ameiurus nebulosus), were collected from three fresh ponds at St. Vincent National Wildlife Refuge, Franklin County, Florida. -Composite-whole-body samples of each were analyzed for mercury and selenium. Mercury concentrations likely to be present in the edible portion (fillet) of these species were estimated from the whole-body value. Largemouth bass ranged in length from 197 to 460 mm (7.7-18.1 in.). Bluegill were from 145 to 240 mm (5.7-9.4 in.) in length, and brown bullhead .were 302 to 363 mm (11.9-14.3 in.) long. Mercury concentrations in all species were below 0.25 mg/kg wet weight, except for bluegill (0.27) in Pond. Selenium may, in some cases, mitigate the adverse biochemical and/or physiological effects of mercury. Selenium has also been found to be toxic to wildlife in large amounts. Selenium concentrations in all composite samples were < = 0.2 mg/kg, wet weight. Neither mercury nor selenium were found to occur at levels believed to be toxic to individual fish or consumers. However, * the mercury concentrations found in the fish may constitute some risk to predators that regularly consume them.

KEY WORDS: mercury, selenium, largemouth bass, bluegill, brown bullhead, St. Vincent National Wildlife Refuge CONTENTS '

ABSTRACT i CONTENTS' ...:. ii FIGURES . . .. , . .'..; lit TABLES . . ;:...... v ...... iii APPENDICES ...... iii ACKNOWLEDGMENTS , iv INTRODUCTION .....' .' 1 SITE,DESCRIPTION .".', . . .'. 2 SAMPLING STATIONS , .' . ...,. . .' 4 MATERIALS AND METHODS . , 4

RESULTS AND DISCUSSION f ...... 6 CONCLUSIONS AND-RECOMMENDATIONS 12 LITERATURE CITED . .". . 13 APPENDICES 19 FIGURES

Figure 1. Location of St. Vincent NWR 3

Figure 2. Fish collection stations at St. Vincent NWR 5

Figure 3. Mean mercury in fish composite samples by average length 8

Figure 4. Meahliiefcury in fish composite samples from three ponds at St. Vincent-NWR 8

Figure 5. Mean selenium fish composite samples by average length 10

Figure 6. Mean selenium in fish composite samples from three ponds at St. Vincent NWR , 10

TABLES

Table 1-. Composite fish samples, St. Vincent NWR, 1988 : 7

Table 2. Mercury concentrations in composite samples of fish from the St. Vincent NWR,1988 9

APPENDICES

Appendix A. The Nature of Mercury 15

Appendix B. The Nature of Selenium 20

Appendix C. Interactions of Mercury and Selenium in Biota T 23

Appendix D. St. Vincent NWR Sampling Station Locations 25

Appendix E. Standard Operating Procedures for <• Collection of Fish Tissue Samples 26

Appendix F. Laboratory QA/QC Procedures 28

Appendix G. Study Data 30

in ACKNOWLEDGMENTS

Fish .samples were collected with the assistance of Johri Odenkirk,-' Biologist, Panama City Field. Office and Kennard Watson, Service volunteer. Logistical support was provided'by Jerry Hoflpma'n, Refuge Manager, St. Vincent National Wildlife Refuge, and his staff, Terry.. .-(*;. . • • •, .. : ' . . .• Phillips, Robert?Gay'and Tom Gay: ..• ,, ' .- . , - './• . . :. *\.&r -V ;• '-'--•' * • : : r~a • , ;

We W&I& .assisted in report preparation;;and review by Dr. Charles Facemir-e, Region 4 Senior Enviroiirnental Contaminants Specialist; Frank Finchum, Kathy Hqffmaster and Danelle Kinion, Office Auto'matioh Clerks; Hildreth Cooper, Biologist; aiid Gail A.. Carmody, 'Project Leader,

Panama. City :Fleld Office. . • •

IV INTRODUCTION

In September 1982, the Florida Game and. Fish Commission conducted a fish survey in the Chipola of northwest Florida to determine if the fishery was contaminated. This action was taken when it was found that a battery salvage .plant-located in Jackson County had released contaminated effluent into__the_Chipola River. Elevated levels of mercury were found a in largemouth bass (Micropterus salmoides) collected from the Dead- area of the River. To obtain background measurements for comparison, fish were collected from the Santa Fe River, which was thought to be relatively pristine. Results were surprising. Santa Fe River bass also contained elevated mercury levels (Bigler et al., 1985). These results led to the formation of an informal interagency task force composed of personnel from the Florida Game and Fresh Water Fish Commission, the Florida Department of Environmental (now the Department of Environmental Protection), and the Florida Department of Health and Rehabilitative Services (HRS). Subsequently, a systematic statewide mercury investigation was initiated that involved the sampling of about 20 Florida lakes or streams each year.. In 1988, this . on-going investigation revealed elevated concentrations of mercury in largemouth bass and other species collected in the Everglades waterways of south Florida.

As a result of the State's mercury investigation, fish consumption health advisories were formulated by HRS for largemouth bass and other species. The advisories recommend that when the average concentration of mercury in the edible portion (i.e., fillets) is between 0.5 and 1.5 ppm wet'weight, healthy should limit their consumption to no more than one meal (=4 . oz.

The principle objectives of the NWR studies were to determine if fish had levels of that might be injurious to individuals or populations of fish and wildlife species under'refuge management, and sufficient to trigger issuance of human health consumption advisories for the refuges. .

SITE DESCRIPTION

St. Vincent National Wildlife Refuge is a 12,358-acre undeveloped barrier island located just south of the mouth of the Apalachicola River in Franklin County, Florida (Figure 1). The island is dissected by dune ridges. Many of the sand roads on St. Vincent follow these ridges, extending from east to west the length of the island. The interdune habitat areas vary from fresh water lakes and sloughs on the eastern end to dry upland pine forests on the western end of the island. The. climate is mild and subtropical with an average annual rainfall of 57 inches. Four miles wide at the east end and nine miles long, this triangular island is larger and wider than most of the northern Gulf Coast barrier islands-.

o • t> The Refuge preserves highly varied plant and animal communities. Ten separate habitat types have been identified: wetlands, consisting of tidal marsh and fresh water lakes and streams; dunes dominated by live oak/mixed hardwood overstory, scrub oaks, or live oak/scrub oak mix; relatively pure stands of cabbage palm; and four different slash pine communities, each with its own unique understory species. ^ ^ - .-.--.. •- • •-. : •XO«ti "•'•/••'• • ' '-S^iStH*''

V?*-!i$XS<:,;$&*.";••;• .':;..i-^Sil =»Sff» ^;p;^-p^ bSfS3§i2uS4£/£Ir7^r •^•f^JSp*'-

. VINCENT N.W.R. -%?'?-'.-VC•.• •«.-• - < -v-^;?-;- "•• v-- .•;*^y- --aft ' ' -"'*

Figure 1. Location of St. Vincent National Wildlife Refuge Initially, the Refuge was established for waterfowl, but its mission has been broadened to include the protection of habitat for endangered species and to provide a variety of recreational activities. It provides sanctuary for a number of endangered and threatened species. Bald nest in pines near the fresh water lakes and marshes. Loggerhead sea turtles come ashore to nest on the pristine beaches. Indigo snakes inhabit gopher tortoise burrows in the dunes. The Refuge is a resting area for wood storks and peregrine falcons.

In 1990, St. Vincent National Wildlife Refuge became, onejof several southeastern coastal islands where endangered red wolves are being bred. After the wild pups have been weaned, .they are taken to reintroduction sites such as Alligator River National Wildlife Refuge in North Carolina and Great Smoky Mountains National Park.

SAMPLING STATIONS

Fish were collected from three fresh water ponds (Figure 2). The Refuge ponds varied in size: Pond 2 = 31 acres, Pond 4 = 30 acres, and Oyster Pond = 116 acres. .Average depths were 3-4 feet with maximum depths of 6-8 feet. Extensive stands of emergent and submergent aquatic vegetation were found throughout the shallow littoral zone of the ponds. Coontail (Ceratophyllum demersim), bladderwort (Utricularia spp), widgeon grass (Ruppia maritima), musk-grass (Chora spp) and cattails (Typha spp) were the dominant vegetation types found around the ponds. Pond bottoms varied from oyster shell composition to sand, mud, or heavy organic'. Most of the pond were brownish-red, tannin-stained waters which were slightly acidic (pH=6.3-6.9). (Parauka 1987). Appendix D contains latitudinal and longitudinal coordinates for each station. • t>

MATERIALS AND METHODS

Fish were collected with gill nets or by hook and line.. Collected specimens were immediately

placed on ice in clean thermal containers. Upon ^-return-ing'-to Panama City:,*samples'were prepared in accordance with standard operating procedures (Appendix E) and placed in a storage freezer maintained at -23°C(-10°F). Samples were shipped to the analytical laboratory after St. Vincent Sound • •• ' -fe.'i" ' • ''•',>

. •«': .!t, JtUii? '" B.

.St. Vincent Island f ''^ '•jr-;'.-,;. y.

• \'- •'•'"•• '•"$'•'. North V M •-•/, -X i:-^ \Pond 5 (&. \ H -Vj.. . dirtjoad, "

vt!.; ;-e /

Figure 2. A); St. Vinpent National Wildlife Refuge B) Fisn Collection Stations: Pond 2, Pond 4, Oyster Pond approximately 30_ days of freezer storage. Laboratory protocols are found in Appendix F. Appendix G contains the study data,

RESULT4 S AND DISCUSSION

Analyses of mercury and selenium were performed on composite samples of whole fish. For this reason, the results of this refuge survey are not directly comparable with the other refuge studies for fish/mercury hi Florida. In the other studies, analyses were performed using muscle tissue (fillets) of individual fish. Currently there is no model that allows acceptable estimates to be made of the mercury that would exist in fillet, based on whole-body analyses. The fact that'the samples were composites further complicates any attempt to estimate amounts of mercury that may exist in the fillets of St. Vincent fish. However, a set of data obtained from the Florida Game and Fresh Water Fish Commission enabled crude estimates of mercury concentrations to be made. The data consisted of mercury values-from whole fish and fish fillets obtained from groups of fish of the same length (Lange 1994). These data seem to indicate that the whole body value represents about 65% of the concentration amount that would exist in the fillet alone.

* The three species of fish collected were largemouth bass (Micropterus salmoides), bluegill (Lepomis macrochirus), and brown bullhead (Ameiurus nebulosus). Table 1 provides information relative to the composite samples for this study. The whole-body, composite-sample mercury concentrations (Table 2) ranged from 0.12 to 0.27 ppm, wet weight (ww).

If whole-body values do indeed represent about 65 % of the mercury concentration that occurs in bass fillets, then the amounts occurring in the edible portion of fishes from the Refuge would p still be well below the lower Florida consumption advisory level of 0,5'ppm ww. As reported in Figure 3, largemouth bass and brown bullheads generally acquired more mercury as the fish increased in length. This trend did not hold for bluegill. Smaller bluegill, all from Oyster Pond, contained more mercury than did bluegill from ponds 2 and 4. Table 1. Composite fish samples, St: Vincent National Wildlife Refuge, 1988.

Collection . Sample Sample Total Total Composite Date No. ID " Lengths Weights Weight (mm) . (gm) (gm)

5/6/88 ;,' ._. 1 LMB-P2 353 555 390 '750 .: ' 340 543 •. .315 400

{ 4 "" 2/16/88' „- 2 LM3-P4 "460 1325 434 1065 ••-. - 442 1375 ' . 349 545 f

' '408 925 .... '•5335 ' '

2/18/88' 3, LMB-OP" :' ,201 105. • .>'; '>' •'- 223. 145 - ' .;. 222 155 ',r' 218 135 , .: ; .- .' .' . 197 95 ' ...... v.635 . ;•

2/28/88 ' .""-'. 4 ' *"* 145 ' ,..- -,^^ !•.•':-.<.-.'-.&*•:.."• 165 op^r;"- ""r ••••'184' • •— 212 • 20*0 . 215 ; 190 825

; T : '5/5/88 -.-- J ''*•',. :- -5,;f: • • 7" .' ^~\•' ' i •••J'v-4*BG~P4: " ' ''".•;"•,,--' ( . ' ~"2i5" 'i's./»•-. - "' , ' ; --23.8; .t,);.- "215 280 220 . 260 240 322 210 240 • .; 1340

:• 5/6/88 . •,'. 6 BG-OP . 185 170 160 '100 145 70. 160 100 170 106 546 ;' -'"•,

. 342- 623 360 652 '••'•" -. 327 548

• -•> '" • • 363 657 2950 ' .- , i!

5/5/88 - * 8 CF-E4 . 305 490 355 650 335 570--- 1710. ' t_r.

5/6/88 « "9 Cp-OP ' 350 586'- •X ' 4 ">, 355 -' ' 640 ^ ^ ** 360 x ". 570 *' ' 678 , • ' - '- 340 610 •*- ~ ,. 30§4 : v

LMB=largemouth bass B=bluegill , CF= P4=pond 4 " — -0P=Oyster Pond=== t ~ * ,a~* ^ ~ I 0.5 I 0.4 I £ "5 ~ 0.3 3

Q. Q. > 0.2 o CU 0.1

164 201 212 220 331 338 350 353 418 Average Total Length in Millimeters Largemouth Bass Bluegill Brown Bullhead

Figure 3. Mean mercury in fish composite samples (n=5, *n=3). Horizontal scale is average length for each sample. 0.5 I

0.4

15 "0.3 3

Q. Q. X 0.2

U OJ 0.1

-. Pond 2 Pond 4 Oyster Pond Largemouth Bass 1 Bluegill Brown Bullhead Figure 4. Mean mercury in fish composite samples {n=5, *n=3) from three-ponds at St. Vincent NWR. Because selenium may decrease the toxic effects of mercury (Eisler, 1985, 1987), concentrations of selenium were measured in an attempt to understand possible relationships between it and mercury in largemouth bass. In the most recent nationwide monitoring of selenium in fresh water fishes, selenium ranged from 0.05 to 2.9 ppm (whole fish, ww) and averaged about 0.6 ppm (Eisler 1985). In another study (Eisler 1987) selenium concentrations in muscle tissue of largemouth bass ranged from 0.05 to 1.7 ppm. Selenium values in St. Vincent Refuge fish ranged from 0.1 to 0.2 ppm ww (Figure 5). Selenium concentrations of bass in this study appear . • « . .3 " " to be within the normal range for the species. However, the range is too small to infer any role that selenium may have played in mitigating the toxic effects of mercury.

Table 2. Mercury concentrations in composite samples of fish from St. Vincent National Wildlife Refuge, February - May, 1988.

Sample Concentration ID Location n (mg/kg wet wt)

Largemouth bass LMB-P2 Pond 2 5 0.23 LMB-P4 Pond 4 5 0.24 LMB-OP Oyster Pond 5 0.18

Bluegill BG-P2 Pond 2 5 0.12 BG-P4 Pond 4 5 0.13 BG-OP Oyster Pond 5 0.27

" Brown Bullhead CF-P2 Pond 2 5 0.12 CF-P4 Pond 4 3 0.12 CF-OP 0 Oyster Pond 5 0.14

The data available from this study do not allow any statistical comparison between the sites sampled. However, examination of Figures 4 and 6 appear to indicate that there is no great difference in the mercury and selenium available in the environments of Pond 2, Pond 4 and Oyster Pond. _-. 0,25

-. 0.2 .c ' O)

0.15 I E a. a E 0.1 D c _0) 0) 0.05

164 201 212 220 331 338 350 353 418 Average Total Length In Millimeters Largemouth Bass Bluegill ICKXxJ Brown Bullhead Figure 5. Mean selenium in fish composite samples (n=5, *n=3). Horizontal scale is average length for each sample. 0.25

0.2

J? 0) <$ 0.15 3 E Q. Q.

E .0,1 _3

_Q) 0) V) 0.05

Pond 2 Pond 4 Oyster Pond |v.v.v.| Largemouth Bass Bluegill £9%l Brown Bullhead Figure 6. Mean selenium in fish composite samples (n=5, *n=3) from three ponds at St. Vincent NWR. Until additional samples of bass and other, fish :at the Refuge can be analyzed for fillets alone, the uncertainty regarding the risk related to consumption of Refuge fish,remains. Large largemouth bass could contain mercury in fillets exceeding Florida advisory levels. A recent bass management plan (Florida Game and Fresh .Water Fish Commission, 1992) requires that: bass caught hi waters north and west of the:Suwannee River be at leastJJZ inches (305 mm) in length. Thus, people particular lakes or ponds on the Refuge should be aware that the- . a ' ~ ' ' ~ only fish they are allowed to take could exceed the State's lower consumption advisory.

There are many reasons why mercury in bass from some geographic locations may be higher than that observed in others. ' D'ltri (1990) has pointed out that the major determinant of the amount of mercury which may be concentrated in a fish is generally the rate of the reaction which converts inorganic (metallic) mercury to organic . Methylmercury, the predominant form found in fish fillets (Luten et al., 1980), is readily b.ioavailable and highly toxic (Eisler 1987). The rate of memylation is dependent upon many.factors including water pH, hardness, alkalinity and conductivity (D'ltri, 1990; Wiener et al., 1990). .

•r

Mercury levels in the whole-body composite samples of bass from this Refuge were well below those reported by Armstrong (1979) to cause either chronic (e.g., loss of appetite, inability to catch , rolling from side to side) or.acute .(i.e., mortality) to the fish themselves. However, concentrations in whole-body samples of all of. the fish were above the level (0.1 ppm) which Eisler (1987) believed to be protective of sensitive species of fish- birds. Reproductive impairment has been,noted in some avian species which regularly.ingested 0.05 to 0.1 ppm of mercury in their diet (Eisler 1987)... . . •

Although total mercury concentrations can sometimes be greater.in whole fish than in fillets, this is usually due to high levels of inorganic mercury in the liver (Luten et al., 1980)." of inorganic mercury occurs at a much slower rate than methylmercury due to its relative inability to penetrate the gills and gastrointestinal tract (Olsen et al., 1973). Thus,

11 adverse effects from inorganic mercury concentrations in whole fish, as consumed by a bird or other predator, -are likely to be minimal when compared to those caused by the methylmercury content of the food item. .. a

Mink (Mustela vison) are reported to be one of the carnivorous species most sensitive to contaminants, including mercury, transported through the aquatic (Wren, 1986). Mercury concentrations in'Refuge bass were much less than 5.0 ppm, the dietary level reported to be lethal to mink by Aulerich et al. (1974). If other carnivorous mammals (otter, raccoon, etc.) are less sensitive than mink, there should be no significant risk to these animals.

CONCLUSIONS AND RECOMMENDATIONS •

The data obtained for mercury and selenium in fishes at St. Vincent National Wildlife Refuge indicate that minimal mercury contamination is present, and that selenium values fall within the normal range for that . The nature of the samples (i.e. composite) and the type of analysis (whole-body) do not allow direct comparison with State of Florida human consumption advisories based on analysis of fillets taken from individual fish. While mammal species are probably not at risk from consumption of fishes on the Refuge, some avian species could experience adverse physiological/biochemical effects. Based on the above71he following recommendation is made:

1. Conduct an evaluation of the fishes of St. Vincent National Wildlife Refuge with analyses of edible fillets rather than whole fish, to assure that any risk to either recreational fishermen or wildlife is minimal.

12 LITERATURE CITED

Armstrong, F.A.J. 1979. Effects of mercury compounds on fish. Pages 657-670 in Nriagu, ed. The biogeochemistry of mercury in the • environment. Elsevier/North Holland Biomedical Press, New York.

Aulerich, R.J., Ringer, R.K. and Iwamoto, S. 1974. Effects of dietary mercury on mink. Archives of Environmental Contamination and Toxicology 2, 43-51.

Bigler, W.J., Ware, P., Savage, T., King, S. and Hartwig, C. 1985. Heavy in fish and from the Chipola and Santa Fe of north Florida. Florida Academy of Science 9pp. .

D'ltri, P.M. 1990. The methylation and cycling of selected metals and metalloids in aquatic sediments. Pages 163-214 in R. Baudo, J.P. Giesy and H. Muntau, eds. Sediments: chemistry and toxicity of in-place pollutants. Lewis Publishers, Chelsea, Michigan.

Eisler, Ronald. 1985. Selenium hazards to fish, wildlife, and invertebrates: a synoptic review. U.S. -Department of the Interior, Fish and Wildlife Service. Biological Report 85(1.5), Contaminant Hazard Reviews Report No.5.

Eisler, Ronald. 1987. Mercury hazards to fish, wildlife, and invertebrates: a synoptic review. * U.S. Fish and Wildlife Service Biological Report 85(1.10). 90pp.

Florida Department of Health and Rehabilitative Services. 1989. Health advisories for Florida waters. Public Information Office, Tallahassee.

Florida Game and Fresh Water Fish Commission. 1992. New bass fishing laws to take effect July 1, 1992. (News release dated March 4, 1992).

Lambou, V.W., Barkay, T., Braman, R.S., Delfino, J.J., Jansen, J.J., Nimmo, D., Parks^ J.W., Porcella, D.B., Rudd, J., Shultz, D., Stober, J., Watras, C., Wiener, J.G., Gill, G., Huckabee, J. and Rood, B. 1991. Mercury technical committee interim report to the Florida Governor's Mercury in Fish and Wildlife Task Force. Environmental Monitoring and Wet Environments Research Program, Florida State University, Tallahassee. 60pp.

Lange, Ted. 1994. Personal communication. Largemouth bass whole-body and fillet mercury data. Florida Game and Fresh Water Fish Commission. Eustis, Florida.

Luten, J.B., Ruiter, A., Ritskes, T.M., Rauchbaar, A.B. and Riekwel-Booy, G. 1980. Mercury and selenium in marine and fresh water fish. Journal of 45, 416-9.

13 I

Olson, K.R., Bergman, H.L. and Fromm, P.O. 1973. Uptake of methyl mercuric chloride by I : a study of uptake pathways into the whole, animal and uptake by erythrocytes in vitro.. Journal of the Board of 30, 1293-9.

Parauka, Frank. 1987. Fishery management plan, St. Vincent National Wildlife Refuge. U.S. Fish and Wildlife Service. Panama City Field Office, Florida.

Sokal, R.R. and Rohlf, FJ. 1969. Biometry:- the principles and practice of statistics in biological research. W.H. Freeman and Company, San Francisco. 776pp.

Wiener, J.G., Martini, R.E., Sheffy, T.B. and Glass, G.E. 1990. Factors influencing mercury concentrations in walleyes in northern Wisconsin lakes. Transactions of the American B Fisheries Society 119, 862-70.

Wren, C. 1986. A review of metal accumulation and toxicity in wild mammals: I.Mercury. D Environmental Research 40, 210-44. •

14 APPENDIX A

THE NATURE OF MERCURY

^ ' 4 . Mercury (Hg) and its compounds have no known normal metabolic function. The presence of mercury in cells of living organisms represents contamination from either natural or anthropogenic sources, or both. Mercury contamination at the cellular level should be regarded . a as undesirable and potentially hazardous (Eisler 1987). •• •°~

Some forms of mercury with relatively low toxicity can be transformed into forms with very high toxicity through methylation by various biotic and abiotic processes. Methyl mercury can be bioconcentrated in organisms and biomagnified through food chains, returning mercury directly to .man and other upper consumers in concentrated form. Mercury has mutagenic, teratogenie"and carcinogenic -properties, and has caused embryocidal, cytochemical and histopathological effects. High body burdens of mercury normally encountered in some species of fish and wildlife from remote locations emphasize the complexity of natural mercury cycles and human impact on these cycles. Some scientists believe that the anthropogenic release of mercury into the environment should be curtailed because the difference between tolerable natural background levels of mercury and harmful effects in the environment is exceptionally small (Eisler 1987). - - • •

Mercury from natural sources can enter the biosphere as, a gas from terrestrial and oceanic volcanic activity, in solution or in particulate form. Cinnabar (HgS) is a common mineral in hot springs deposits and a major natural source, of mercury. The global cycle of mercury involves degassing of the element from the. earth's crust, evaporation from natural bodies of water, atmospheric transport (mainly in,the form of mercury .yap or), and wet or dry deposition-of mercury back onto land and water. Oceanic effluxes of mercury are tied to equatorial and phytoplankton activity and may significantly affect the global. cycling of this metal. If volatilization of. mercury is proportional to primary production in the world's , oceanic

15 phytoplankton activity represents about 36 percent of the yearly mercury flow to the (Eisler 1987).

Human activities that contribute significantly to the global input of mercury include the combustion of fossil fuels, and reprocessing of gold, copper, and , operation of chjor- alkali plants, and disposal of batteries and fluorescent lamps. The production of electrical "apparatus, industrial control instruments (switches, thermometers, and barometers, etc.), laboratory appliances, anti-fouling and mildew-proofing , chemical formulations to control fungal of seeds, bulbs, and vegetables, dental amalgams, pulp and 'paper, Pharmaceuticals, and metallurgy and mining, is contributing, or has contributed, mercury to the environment (Eisler 1987). . . I Mercury burdens in sediments and other non-biological materials are estimated to have increased ~ up to five times prehuman levels; primarily as a result of man's activities. The estimated half- time residence value for mercury is comparatively short in the atmosphere, between 6 and 90 days, but is much longer in terrestrial „ oceanic waters, and oceanic sediments where it is '• estimated to remain 1,000, 2,000, and more than one million years, respectively (Eisler 1987).

An elevated concentration of mercury (usually as methyl mercury) in any biological sample is often associated with proximity to human use of mercury. The elimination of mercury point- source discharges has usually been successful in improving environmental quality. However, elevated levels of mercury in biota may persist in contaminated areas long after the source of - t> has been discontinued. It is noteworthy that some groups of organisms with consistently elevated mercury residues may have acquired these concentrations as a-result of natural processes, rather than from anthropogenic activities. These groups include older specimens of long-lived predatory fishes, marine mammals (especially seals and sea lions), and

organisms living near natural-mercury:-'oreMnnabar deposits, .. s-'.'V->-^-v->--:-v ; - • -^^•^•

16 I Certain species of macrophytes strongly influence mercury cycling. For example, Spartina alterniflora, a dominant marsh plant in Georgia estuaries, accounted for almost half the total mercury budget in that (Eisler 1987). Mangrove vegetation plays a similarly important role in mercury cycling in the Florida everglades (Eisler 1987). These findings suggest that more research is needed on the role of higher plants in the -. In. aquatic , removal of the source of anthropogenic mercury results in a slow decrease in the mercury content of sediments and biota. The rate of loss depends, in part, on the initial degree of contamination, the chemical form of the mercury and the half-life of that form, physical and chemical conditions of the system, and the hydro-dynamics of the particular . " .

Methyl mercury is produced by methylation of inorganic mercury present in both fresh water and saltwater sediments, and accumulates in aquatic food chains in which the top level predators usually contain the highest concentrations (Eisler 1987). Most organomercury compounds other than methyl mercury decompose rapidly in the environment, and behave much like inorganic mercury compounds (Eisler 1987). In organisms near the top of the food chain, such as carnivorous fishes, almost all mercury accumulated is in the methylated form, primarily as a result of the consumption of prey containing methyl mercury. A strong relationship appears, to exist between elevated mercury in Florida largemouth bass and low pH waters from swamp or peat drainage. A negative correlation exists in Florida for highly eutrophic (enriched) waters^ where depressed mercury levels are typically found. " Methylation also occurs within the biological organisms themselves because intestinal convert mercury into methyl mercury through enzymatic processes. However, this methylation process, as a mercury uptake source, is not as important as intake of methyl mercury via the animal's diet.

There is no known effective antidote to counteract the effects of methyl mercury on the vertebrate central nervous system (Eisler 1987).- Mercury binds strongly with sulfhydryl

17 groups and has many potential target sites during embryogenesis. Phenyl mercury and methyl mercury compounds are among the strongest inhibitors of cell division (Eisler 1987). Organomercury compounds, especially methyl mercury, cross placental barriers and can enter mammals by way of the respiratory tract, gastrointestinal tract, skin or mucous membranes (Eisler 1987). Compared with, inorganic mercury compounds, organomercurials axe more completely absorbed, or more soluble in organic solvents and lipids, pass more readily across

. a biological membranes, and are slower to be excreted (Eisler 1987)7 ~

Mercury, at comparatively low -concentrations, adversely affects the reproduction, growth, behavior, , chemistry, osmoregulation, and oxygen exchange of marine and fresh water organisms (Eisler 1987). In general, the accumulation of mercury by aquatic biota is rapid, and depuration is slow.- Organomercury compounds, especially methyl mercury, have been found to be significantly more effective than inorganic mercury compounds in producing adverse effects and accumulations. Adverse affects of mercury to aquatic organisms have been documented at water concentrations of 0.88 to 5.0 ug/1. Enzyme disruption occurred in brook trout (Salvelinus fontinalis) embryos exposed for 17 days in solutions containing 0.88 ug/1 of methyl mercury (Eisler 1987). Increased incidence of frustule abnormalities and burst thecae

were documented in two species of marine exposed to 1.0 ug/1 concentrations of Hg++ for 24 hours (Eisler 1987). Arrested development of larvae occurred in a 40-hour test

when the larvae were exposed to 3.0_ug/l concentrations of Hg++ (Eisler 1987). Decreased rate of intestinal transport of glucose, fructose, glycine, and tryptophan occurred in the murrel7

Channapunctatus, when exposed to 3.0 ug/1 concentrations of Hg++ for 30 days (Eisler 1987). The blood chemistry of (Morone saxatilis) was altered^when these fish were exposed

to 5.0 ug/1 concentrations of Hg++ for 60 days (Dawson 1982). Decreased respiration in striped bass was .observed 30 days post exposure after immersion for 30 to 120 days in 5.0 ug/1

concentrations of Hg++ (Eisler 1987).

18 The environmental cycle of mercury is delicately balanced and small changes in input rates, and/or the chemical forms of mercury, may result in increased methylation rates in sensitive systems. For example, the acidification of natural bodies of fresh water is statistically associated with elevated concentrations of methyl mercury in the edible tissues of predatory fishes. In chemically sensitive waterways such as poorly, buffered lakes, the combined effects of acid precipitation and increased emissions of mercury to the atmosphere (with subsequent deposition) . a , a pose a serious threat to the biota if optimal biomethylation conditions are met.

LITERATURE CITED Dawson, M.A. 1982. Effects of long-term mercury exposure on hematology of striped bass, Morone saxatilis. U.S.. National Marine Fisheries Service, Fisheries Bulletin 80, 389-92.

Eisler, Ronald. 1987. Mercury hazards to fish, wildlife, and invertebrates: a synoptic review. U.S. Fish and Wildlife Service Biological .Report 85(1.10). 90pp.

I

19 APPENDIX B THE NATURE OF SELENIUM

All investigators appear to agree on four points. First, that insufficient selenium in the diet may have harmful, and sometimes fatal, consequences. Second,,that exposure to grossly elevated levels of selenium in the diet or water, is inevitably fatal over time to terrestrial and aquatic organisms. Third, that there is a comparatively narrow concentration range separating effects of selenium deficiency from those of selenosis. Fourth, that additional fundamental and basic research is required on selenium metabolism, physiology, recycling, interactions with other compounds or formulations, and chemical speciation in order to elucidate its nutritive role, as well as its toxic effects. Accordingly, the proposed selenium criteria-for prevention of selenium deficiency and for protection of aquatic life, livestock, crops, and human health, should be viewed as guidelines, pending acquisition of additional, more definitive data......

Selenium chemistry is complex. In nature, selenium - exists as six stable isotopes, three allotropic forms, and in five valence states. .

Selenium , a non-metallic element, occurs naturally in the environment in trace amounts and rarely exceeds 2 ppm dry weight in soils. Selenium is an essential micronutrient for normal animal nutrition, but concentrations exceeding ;those required may produce toxic effects "ranging from physical malformations during embryonic development to sterility and . TwcTmajor sources of selenium are agricultural irrigation return-flows that originate from high selenium soils, and drainage water from areas used for storage and disposal of ash produced by -fired power plants.. , . . • .

Because selenium in aquatic systems is readily taken up by organisms, concentrations sometime reach levels toxic to fish and wildlife. Three things can happen to dissolved selenium when it enters an ecosystem: 1) it can be absorbed or ingested by organisms, 2) it can bind or complex

20 with particulate matter, or 3) it can remain free in solution (Lemly and Smith 1987). Through deposition of biologically incorporated selenium and settling of particulate matter, selenium accumulates in the top layer of and detritus. Biological, chemical, and physical processes move selenium out of, as well as into, the sediments. The sediments are only a temporary repository for selenium. Aquatic systems are dynamic and. selenium can be cycled back into the biota and remain at elevated levels for years after waterborne inputs of selenium are stopped. •a- ~~~

Selenium may be removed from solution and held in sediments through the natural processes of chemical and microbial reduction of the selenate form (Se VI) to the selenite form (Se IV) followed by adsorption onto clay and the organic carbon phase of particulates, reaction with species, and co-precipitation or settling. Immobilization processes effectively remove selenium from the soluble pool, especially in slow-moving or still-water habitats and wetlands (Lemly and Smith 1987). " .

Selenium in sediments is particularly important to long-term habitat quality because mechanisms in aquatic systems can mobilize selenium into food chains, and thereby cause long-term dietary exposure of fish and wildlife when it is made available for biological uptake by oxidation and methylation processes.

The aquatic systems that accumulate selenium most efficiently -are shallow, standing or slow- moving waters that have low flushing rates. Several of these habitat types often occur together in one aquatic system. Rivers may have fast-flowing waters, slow moving pools and standing- backwater areas, all within a few hundred meters. The degree of fish exposure to selenium varies among habitats according to intensity of use, type of use, and relative contributions of the various processes that regulate selenium cycling.

21 Selenium is chemically similar to sulphur and because it is an essential micronutrient, extensive bioaccumulation may result. of selenium (the accumulation of progressively higher concentrations by successive trophic levels of a food chain) usually ranges from 2,to 6 times between the producers (algae and plants) and the lower consumers (invertebrates and ) (Eisler 1985)._ Top level consumers, such as , may receive toxic selenium levels in the diet even though the concentration in water is low. The risk of toxicity through the detrital food pathway will continue despite a loss of selenium 'from the , as long as contaminated sediments are present.

Toxic effects of selenium fall into two categories: 1) mortality of juveniles and adults, and 2) reproductive effects (Lemly and Smith 1987). Complete reproductive failure can occur with' little or no tissue pathology or mortality in the population. Field and laboratory data suggests that selenium at concentrations greater than 2-5-parts per billion (ppb) in water can be bioconcentrated in food chains and cause toxicity and reproductive failure in fish. Selenium may interact with several metals that can alter the expression of biological effects. Other factors such as temperature, nutrition , differences in species sensitivity, differences in the relative toxicity of the various chemical forms of selenium and other environmental stresses may affect the actual concentration of selenium that produces toxicosis.

LITERATURE CITED Eisler, R. 1985. Selenium hazards to fish, wildlife, and invertebrates: a synoptic review. U.S. Department of the Interior, Fish and Wildlife Service. Biological Report 85(1.5), Contaminant Hazard Reviews Report No.5.

t> Lemly, A.D. and GJ. Smith. 1987. Aquatic cyucling of selenium; implications for fish and wildlife. Fish and Wildlife Service Leaflet 12, U.S. Fish and Wildlife Service.

22 APPENDIX C INTERACTIONS OF MERCURY AND SELENIUM IN BIOTA

The protective action of selenium against the adverse or lethal effects induced by various metals and metalloids'is well documented-for a wide variety of plant and animaLspecies; however, not all tests were conclusive. Studies with some species of fresh water teleost fishes demonstrated

- a negligible antagonism of selenium against mercury (Eisler 1'985).

Reasons to account for the antagonism of selenium and mercury (as well as other metals) include dietary source and chemical form of selenium, influence of sulfur, biological translocation of selenium or mercury to less critical body parts, and chemical'linkage of ' selenium to mercury on a linear basis. The exact mode of interaction is probably complex and has not yet been resolved. ..In regard to diet, selenium of animal origin and in the form of selenate is less effective than selenium from plant and inorganic sources in preventing methylmercury in experimental animals (Eisler 1985). Disruption -of sulfur metabolism by selenium, the sulfur being replaced by seleno-amino acids and other cell constituents containing selenium in living organisms, is one probable cause of selenium poisoning. It is conceivable that selenium-mercury compounds formed within the organism would be sufficiently nonreactive biologically to interfere with-sulfur kinetics, presumably -SH groups (Eisler 1985, 1987). Differential redistribution of selenium or mercury to less critical body parts may partly account for observed antagonisms. Pretreatment of marine minnows with selenium protects against and causes a marked redistribution of mercury among organs, presumably to non-criticaj'body parts, and this transfer may partly account for the observed selenium-mercury antagonisms in that species (Eisler 1985). Some investigators have reported that selenium results in increased mercury accumulations. Increased retention of mercury and other metals may lead to a higher-level of biomagnification in the food chain and higher body burden in the individual, which might counteract the positive effect of decreased

23 intoxication (Eisler 1987). Extensive research is under way on the chemical linkage of selenium and mercury (Eisler 1985).

e LITERATURE CITED Eisler, Ronald. 1985. Selenium hazards to fish; wildlife, and invertebrates: a synoptic review. U.S. Department of the Interior, Fish and Wildlife Service. Biological Report 85(1.5), Contaminant Hazard Reviews Report No.5.

Eisler, Ronald. 1987. Mercury hazards to fish, wildlife, and invertebrates: a synoptic review. U.S. Fish and Wildlife Service Biological Report 85(1.10). 90pp.

24 APPENDIX D

j. V ST. VINCENT NATIONAL WILDLIFE REFUGE

SAMPLING STATION LOCATIONS

Station- •;':/; • , - • No, ... Latitude Longitude Location Name.—

1 ...*".- 29°38'30" 85°06'42" . Pond 2

•2 - "' 29038'1-0" 85°07'06" ' Pond 4

3'"-:'.'" 29038'20n 85°08'30" Oyster Pond

25 I APPENDIX E

STANDARD OPERATING PROCEDURES COLLECTION OF FISH-TISSUE.SAMPLES PCFO-EC-SOP-001

Fish collected for chemical contaminant evaluations may be taken by electrofishing gear, monofilament gill nets, otter trawl, haul or beach seines, fish traps, trotlines, or rod and reel. "However, any collecting gear should be free of chemical treatments and/or metals that copld contaminate samples. This is particularly important when the entire fish (whole-body analysis) will be used.

For species of special concern such as Gulf or large broodstock striped bass, we utilize only incidental mortalities, and these should be fresh specimens.

The following is for sample dissection:

1. Wash hands thoroughly and rinse completely. Wear vinyl or latex gloves. Final rinse with distilled water. " - - •

2. Fish should be clean. It may be rinsed of debris or mud in the waters of the collection site. . * » 3. The dissection surface (work area) should be a chemically inert substance such as a stainless steel acetone-rinsed pan, or counter. Avoid letting the dissected sample touch this surface, if possible.

4. Use previously cleaned, and acetone-rinsed, then distilled water-rinsed stainless steel dissection tools (knives, scalpels, etc.). Scales for total fish weights and sample weights should also be clean or covered with pre-cleaned aluminum foil. Measuring devices for fish lengths, etc., should be clean, or should not come in contact with the specimen.

5. Do not let dissected samples remain exposed fo the air^. Exposure.can dry samples and reduce the natural percentage of moisture. Prepare each dissected sample for shipping or freezing as it is dissected.

6. Samples should be placed in the smallest, pre-cleaned glass jar that will adequately hold the sample. The jars should be pre-labeled with a permanent, waterproof marking pen on the outside of the jar. Jars should also have a teflon liner inside the lid. As an alternative, acetone-rinsed, heavy-duty aluminum foil

26 may be used to wrap the sample. After double-wrapping, place the sample (with sample identification label) inside an air-tight zip-lock bag.

7. Sample identification labels should be prepared with permanent, waterproof ink or other writing instruments that will not bleed out or wash out, and should ' provide the following information:

a. species name and common name, b. type of tissue (if not whole-body), c. collection location, .' '- d. latitude and longitude, e. county and state, f. weight of sample in grams, g. date of collection-; h. sample collector's name, i. total weight of fish specimen (grams), j. total length and fork length of specimen (mm), and k. method of collection. •

8. Samples should be frozen as soon as possible. If samples contain large amounts of liquids that may expand, the lids may be set on the jars, without securing, until the sample has expanded and frozen. The lids should then be secured tightly. •F 9. Photographs of the specimens are desirable, as well as a written description of any external or internal lesions, tumors, etc.

27 0-: ;iAPPENDIX.

•A

FOR MERCURY ANALYSIS

28 U. S. FISH AND WILDLIFE SERVICE . . _ Q , - , PATUXENT ANALYTICAL CONTROL FACILITY • '[j D1" ' QUALITY ASSURANCE- REPORT

RE: 5651 REGION: 4 REGIONAL ID: 88-4-118 THE ANALYSES ON THE ABOVE MENTIONED SAMPLES WERE PERFORMED AT: HAZLETON LABORATORIES AMERICA, INC. 3301 KINSMAN BLVD. __ MADISON, WISCONSIN 53704

AFTER A THOROUGH REVIEW OF THE REPORTS ISSUED BY THE LABORATORY, I REPORT THE FOLLOWING OBSERVATIONS AND CONCLUSIONS: THE ACCURACY, AS MEASURED BY SPIKE RECOVERY AND REFERENCE- MATERIAL ANALYSIS, WAS ACCEPTABLE" FOR ALL ANALYTES. THE PRECISION, AS MEASURED BY DUPLICATE SAMPLE ANALYSIS, WAS ACCEPTABLE. WE HAVE NOT RECEIVED SUFFICIENT DATA FROM THIS LABORATORY TO ESTIMATE CONFIDENCE INTERVALS.

...... QUAMTY ASSURANCE OFFICER DATE

I Environmental Trace Substances Research Center

Route 3 Columbia, Missouri 65203 UNIVERSITY OF MISSOURI . Telephone (314) 882-2151

NITRIC REFLUX DIGESTION FOR MERCURY Approximately 0.5 g. of sample was weighed into a freshly cleaned 50 ml. round bottom flask with 24/40 ground glass neck. For waters, 10 ml. of sample were measured into the flask. Five ml. of concentrated sub-boiled HNCu were .added .and the flask was placed under a 12 inch water-cooled condenser with water running through the condenser. The heat was turned up to allow the HNO-, to reflux no more than 1/3 the height of the columns. Samples were allowed to reflux for two hours. Then the heat was turned off and the samples 'allowed to ' cool. The condensers were rinsed with 1% v/v HC1 and the flasks removed. The samples were diluted with 1% v/v HC1 in a 50 ml. volumetric flask and then transferred to clean, labeled, 2 oz. flint glass.bottles.

COLUMBIA KANSAS CITY ROLLA ST. LOUIS IP Environmental Trace Substances Research Center III Roule 3 • • • Columbia, Missouri 65203 UNIVERSITY OF MISSOURI Telephone (314) 882-2151

NITRIC - PERCHOLORIC DIGESTION - (ICP) . a Approximately 0.5 g. of sample was weighed into a freshly cleaned 100 ml. quartz Kjeldahl flask. (Sediment samples and samples containing a high percent of silica were digested in 100 ml. teflon beakers.) For water samples, 50 ml. of sample were measured into a teflon beaker. Slowly 15 ml. of concentrated sub-boiled HNCL and 2.5 ml. of concentrated sub-boiled HC10, were added. Foaming may .occur with some samples. If the foaming started to become excessive, the container was cooled in a beaker of cold water. After the initial reaction had subsided, the sample was placed on low heat until the evolution of dark red fumes' had ceased. Gradually, the heat was increased until the HNO^ began refluxing, samples were allowed to reflux overnight. (This decreased the chance for charring during the reaction with HC10,.) After the refluxing, the heat was gradually increased until the HNCL had been driven off, and thlTreaction with HC10. had occured. When dense white fumes from the HC10, were evident, the samples were removed from the heat and

allowed to cool. Two ml. of concentrated p sub-boiled HC1 were added. The flasks were replaced on the heat and warmed until the containers were hot to the touch or started to boil. They were removed from the heat, and 5-10 ml. of deionized water were added. . Samples were allowed to cool. They were then diluted using deionized. water in-a 50 ml. volumetric-flask and transferred to "clean, labeled, 2 oz. polyethylene bottles.

COLUMBIA KANSAS CITY ROLLA ST. LOUIS

an eo«ai ofjponun.1, r s'-i-'.-O" Environmental Trace Substances Research Center Route 3 Columbia, Missouri 65203 UNIVERSITY OF MISSOURI . Telephone (314) 882-2151

MERCURY - COLD VAPOR ATOMIC ABSORPTION Equipment used for Cold Vapor Atomic Absorption include: Perkin-Elmer Model 403 AA; Perkin-Elmer Model 056 recorder; Technicon Sampler I; Technicon Pump II; a glass cell . o • ; * •••"" \ with quartz windows and capillary tube for entry and exit of the mercury vapor; and a liquid-gas separator. The samples were placed in 4 ml. sample cups at least 3/4 full. • The samples were mixed with hydroxylamine for preliminary reduction, then stannous -•• chloride for reduction to the mercury vapor. The vapor was separated from the liquid and passed through the cell mounted in the light path of the burner compartment.. The peaks were recorded and the peak heights measured. The standardization was done with at least 5 standards in the range of 0 to 10 ppb. The correlation coefficient was usually 0.9999 or better and must have been at least 0.999- to have been acceptable. A standard was run *• every 8-10 samples to check for drift in the standardization. This was usually less than

5%. . Standards were preserved with 10% v/v HN03, 1% v/v HC1 and 0.05% w/v K2Cr207. The solution concentrations were calculated and the data entered into the AA calculation program which corrected for blank, dilution, sample weight, sample volume and entered the data into the LIMS system for report generation"

COLUMBIA KANSAS CITY ROLLA ST. LOUIS Environmental Trace Substances Research Center Rouie 3 n Columbia, Missouri 65203 UNIVERSITY OF MISSOURI Telephone (314) 882-2151

ARSENIC AND SELENIUM BY HYDRIDE

The Varian VGA-76 hydride generation accessory was mounted on either a Perkin-Elmer- • a Model 603 AA or Model 3030" (B) AA. ElectrodelesTDischarge lamps (EDL) were used. The instrument and EDL settings were taken from the instrument manuals. The burner mount for a Perkin-Elmer Model 10 Hydride generator was modified slightly to hold the Varian quartz cell. The cell was aligned in the light path of the burner chamber and a very lean flame was used for heating the cell. The two stock solutions were 50% V/v sub-boiled HC1 and

0.6% NaBH^ in 0.5% NaOH for Selenium and concentrated sub-boiled HCL and 'l% NaBH4 in 0.5% NaOH for . Samples were diluted with 10% v/v sub-boiled HC1. Standards were prepared by dilution of Fisher 1000 ppm stock with 10% v/v sub-boiled HC1 in the range of 0 to 20 PPB. The instrument was standardized to read directly in PPB using SI = 5.00 and S2 = 20.00. After standardization, the standardization was checked by reading other standards such as 2.00, 10.00 and 15.00 PPB and an instrumental quality control sample with.a known value. If the standards and quality control were acceptable, the detection limit was determined by reading the zero standard 10 times, antf"twTclTThT standard deviation of the mean was used as the detection limit. Samples were analyzed by taking an integrated reading for 3 seconds after the plateau was reached for the sample. This occured approximately 45 seconds after the sample tube was placed in the sample. Standardization was checked every 8-15 samples and approximately 10% of the samples were checked by the method of additions to monitor matrix effects. Matrix effects were usually not significant with the VGA-76. The data was corrected for drift.of the standard curve and entered into the AA calculation pro~grf!h. This program corrected for blank, dilution, sample weight, sample volume and recorded the data in the LIMS database for report generation. •

COLUMBIA KANSAS CITY ROLLA ST LOUIS APPENDIX G

STUDYDATA

30 HAZLETOfN LABORATORES AMERICA, INC. 3301 KINSMAN BLVD., P.O. BOX 7545 REPORT OF ANALYSIS MADISON, Wl 53707 USA Patuxent Analytical Control Facility Catalog No. 5651 U.S. Fish and Wildlife Service Purchase Order No. 85800-88-30140 Patuxent Wildlife Research Center Batch No. 88-4-118 Laurel, MD 20708 Contract No. 14-16-0009-87-007 Date Entered: 07/07/89 Attn: Mr. John Moore Date Printed:_ 11/21/89 Analyte: Mercury .

Sample mq/kq Laboratory Identification Matrix Wet Dry LOP Number LMB-P2 Fish 0.234 ' 0.788 0.025 90700892 LMB-P4 Fish 0.245 0.833 0.025 90700893 LMB-OP Fish 0.181 0.686 0.025 90700894 BG-P2 Fish 0.124 0.458 0.025 90700895 BG-P4 Fish 0.133 0.530 0.025 90700896 BG-OP Fish 0.270 1.06 0.025 90700897 CF-P2 Fish 0.124 0.451 0.025 90700898 CF-P4 Fish 0.120 0.465 0.025 90700899 CF-OP Fish 0.143 0.619 0.025 90700900 SD-P2 Sediment <0.025 <0.043 0.025 90700905 SD-P4 Sediment <0.025 <0.039 0.025 90700906 SD-OP Sediment 0.037 0.091 0.025 * 90700907

I

PHONE (608) 241-4471 FACSIMILE (608) 241-7227 TELEX TLX 703956 HAZRAL MDS UD a COHNlroQ Laboratory Services Company HAZLETOIM LABORATORES AMERICA, INC. 3301 KINSMAN BLVD., P.O. BOX 7545 REPORT OF ANALYSIS MADISON, Wl 53707 USA Patuxent Analytical Control Facility Catalog No. 5651 U.S. Fish and Wildlife Service Purchase Order No. 85800-88-30140 Patuxent Wildlife Research Center Batch No. 88-4-118 Laurel, MD 2070S Contract No. 14-16-0009-87-007 Date Entered: 07/07/89 Attn: Mr. John Moore Date Printed: 11/21/89 Analyte: Selenium

Sample mq/kq Laboratory Identification Matrix Wet Dry LOP Number LMB-P2 Fish 0.2 0.67 0.1 •- 90700892 LMB-P4 Fish 0.1 0.34 0.1 90700893 LMB-OP Fish 0.1 0.38 0.1 90700894 B6-P2 Fish 0.2 0.74 0.1 90700895 BG-P4 Fish 0.1 0.44 0.1 90700896 BG-OP Fish 0.1 0.39 0.1 90700897 CF-P2 Fish 0.1 0.36 0.1- 90700898 CF-P4 Fish 0.1 0.39 0.1 90700899 CF-OP Fish 0.1 0.43 0.1 90700900 SD-P2 Sediment 0.2 0.34 0.1 90700905 SD-P4 Sediment 0.2 0.31 0.1 90700906 SD-OP Sediment 0.3 0.74 0.1 90700907

I

PHON'E (508) 241-4471 FACSIMILE (608) 241-7227 TELEX TLX 703956 HAZRAL MDS UD a COflNINQ Laboratory Services Company