EFFECTS AND FATE OF COPPER IN PONDS, Thesis for the Degree of Ph. ,D. ‘ MICHIGAN STATE UNIVERSITY ALAN WILLIAM McIMTOSH 1 9 7 2 1: TI} :3. A R Y Michigan State University {.2 This is to certify that the thesis entitled EFFECTS AND FATE OF COPPER ‘IN PONDS presented by Alan W. McIntosh has been accepted towards fulfillment of the requirements for __Ph-T)- Jemem Fisheries 8c Wildlife 7' Zléc %%.z , quor professor it ... Date W72 44' * ' fl 7") '3) . «I I3";‘,'. ' Q.~ - :3, 0-7639 f u : 5-, 800K BINDERY INC. LIBRARY BINDEHS smumgr. meme” ,__.__. _._..~..._\_.‘ _, _- III 3 II II III IIIIIIIIII I II 1239 0094 948986 09‘- MAGIC 2 . .NGRf__;* '3’1‘1993 ABSTRACT EFFECTS AND FATE OF COPPER IN PONDS BY Alan William McIntosh The effects and fate of copper in two circular ponds 13.7 meters in diameter were studied after application of copper sulfate. The roles of water, sediment, plants, fish and snails in c0pper dispersal were assessed by determining copper concentrations present in the various components before, during, and up to 130 days after c0pper sulfate dosage. Effects of copper on zooplankton, benthos, and green sunfish (Lepomis gyanellus) populations in the ponds were analyzed; approximate lethal limits for organisms were de- termined in preliminary laboratory bioassays. Much of the copper added to the ponds as l and 3 ppm copper sulfate left solution quickly, probably as carbonate precipitates. In one pond, however, nearly 30% of the c0pper applied remained in the dissolved state after 3 weeks. Organic compounds, such as humic acids, may have complexed the copper and kept it in solution. Sediment was the final repository for copper. Passive accumulation of precipitates and direct adsorption of ionic or complexed copper were the probable mechanisms involved in seiiment accum tagger was rel Aquatic IT rapidly to vex 2300 PP!“ coppe "for. of 3 ppm take of copper limited due I: iesage in one I“ to uptake Green 51.] Copper 19ml 5 SOOI‘. returned tratEd Copper basis after t nor snails c: howeVEI' due A dOSag rotifers , Wh Both Cladoce IIEIS of thi ZOOPlanktOn 1n cladoCere Alan William McIntosh sediment accumulation. Once in the sediment, apparently little c0pper was released back into the water. Aquatic macrOphytes and filamentous algae absorbed copper rapidly to very high concentrations; concentrations of above 2000 ppm copper on a dry-weight basis were reached after addi- tion of 3 ppm COpper sulfate to one pond. Despite active up- take of copper, the role of plants in dispersal of copper was limited due to small total mass. MacrOphytes developing after dosage in one pond had elevated copper concentrations, probably due to uptake from the sediment. Green sunfish and snails both concentrated copper. Copper levels in fish were slightly elevated after dosage but soon returned to background levels. Snails actively concen- trated copper to levels of about 400 ppm copper on a dry-weight basis after dosage with 1 ppm copper sulfate. Neither fish nor snails contributed significantly to copper dispersal, however, due to their small total mass. A dosage of 3 ppm copper sulfate killed cladocera and rotifers, while copepods and ostracods appeared unaffected. Both cladoceran and rotifer populations recovered within 4 weeks of this dosage. At 1 ppm copper sulfate, effects on zooplankton were less evident. Although decreases were noted in cladoceran numbers, species differences existed at this dosage level. No direct toxicity of copper to benthic organisms was noted. In one pond, however, release of H28 and low oxygen :erels followir ieerease in chi :‘Jbers did noI Direct to: icwever, all b gala in one p were greater i in a control p ease in search dosage may ha\ Copper 51 algae at l ppr SICmphytes. in c - IMI aCIIIQVed . AdditiOn Alan William McIntosh levels following decomposition of gh2£a_sp. may have caused a decrease in chironomid numbers, while aquatic oligochaete numbers did not change. Direct toxicity of copper to fish was not observed. However, all but four fish died following decomposition of ghggg in one pond. Relative weight increases of green sunfish were greater in a pond treated with 3 ppm COpper sulfate than in a control pond. An inhibition of reproduction and increased ease in searching out prey in the treated pond following dosage may have caused the greater increases. Copper sulfate proved effective in controlling filamentous algae at 1 ppm and moderately effective at 3 ppm for control of macrophytes. Satisfactory control of planktonic algae, includ- ing Ceratium hirundinella and Trachelomonas sp., was not achieved. Additions of 3 ppm c0pper sulfate to two ponds caused some toxicity to fish-food organisms, decreases in aesthetic quality, and sudden changes in water chemistry. At both 1 and 3 ppm c0pper sulfate, removal of algae and macrophytes was temporary. BF flu , L ir EFFECTS AND FATE OF COPPER IN PONDS BY Alan William McIntosh A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Fisheries and Wildlife 1972 Iwish to icctoral progra treviewing ti tie study; Dr. screwing the Thanks go Clarence Goodn plankton and t I am grat lonPenner f0] SI‘3Cial thank: COOyeratiOn d I dedica such of a bic This fee Fellowship (1 Office Of tht grateful. /\ ACKNOWLEDGMENTS I wish to thank the following for their help during my doctoral program: David Jude for his help in the field and in reviewing the thesis; Joe Ervin for his assistance during the study; Dr. Walter Conley for his review and suggestions concerning the thesis. Thanks go to Drs. Donald McGregor, Harry Yeatman, and Clarence Goodnight for their assistance in identifying zoo- plankton and benthic organisms. I am grateful to Drs. Howard Johnson, Dean Haynes, and ( Don Penner for serving as members of my guidance committee. Special thanks go to Dr. Niles Kevern for his counsel and cooperation during this study. I dedicate this thesis to my wife Barbara, who, while not much of a biologist, is an outstanding wife. This research was supported through a Predoctoral Research Fellowship (No. 5-Fl-WP-26, 436-02) from the Water Quality Office of the Environmental Protection Agency to whom I am grateful. ii LIST OF TABLE 115? OF FIGUI :tcaoaucrror: IXTRODUCTIOE; Physics Chemicé Samplir COPPEr Dynami W CDW'UU) CompOn EFFEC TABLE OF CONTENTS Page LIST OF TABLES O O O O O O I O I I O O O O O O O O O O O V LIST OF FIGURES O I O O O O I O O ‘0 O O O O O O O O O O Vii INTRODUCTION 0 O O O O C O O O O O O O O O O O O C O O U 1 PART I FATE OF COPPER IN PONDS INTRODUCTION. . . . . . . . . . . . . . . . . . . . . . 3 MATERIALS AND METHODS . . . . . . . . . . . . . . . . . 4 Physical Description of Ponds. . . . . . . . . . . 4 Chemical Description of Ponds. . . . . . . . . . . 5 Sampling and Sample Preparation. . . . . . . . . . 14 Copper Analysis. . . . . . . . . . . . . . . . . . 15 RESULTS AND DISCUSSION. 0 O I O O O O O O O O O O O O 0 17 Background Levels. . . . . . . . . . . . . . . . . l7 Dynamics of Copper Added to Ponds. . . . . . . . . 17 water I O O I O O O C O O O O O O O O O O O 0 1? Sediments . . . ... . . . . . . . . . . . . . 31 Plant Tissue. . . . . . . . . . . . . . . . . 38 FiSh. O O I I O O O O O I I O O O O O O I O O 49 snails. O O C O O O O O O O I O O O O O O O O 55 Components of Copper Distribution. . . . . . . . . 55 SUMMRY O O I O O O O O O O O O O O O O O O C O O O I O 63 PART II EFFECTS OF COPPER ON FISH AND FISH-FOOD ORGANISMS INTRODUCTION. . . . . . . . . . . . . . . . . . . . . . 65 iii 3221301? comer: SERIALS AN D I". Laborator§ Pond Stud: FZS'JLTS AND 01' Laborator Bioa Grov Pond Stud Zoo; Ben‘ Fisl HHQRY , . FATE-MAL S Al IGSULTS Ann 31mm 31‘: .. , LTLNCES ‘h a. PE» “DIX. TABLE OF CONTENTS--Continued Page MATERIALS AND METHODS O O C O O I O O O O C O O O C O O 6 6 Laboratory Studies . . . . . . . . . . . . . . . . 66 Pond Studies . . . . . . . . . . . . . . . . . . . 67 RESULTS AND DISCUSSION. . . . . . . . . . . . . . . . . 69 Laboratory Study ... . . . . . . . . . . . . . . . 69 Bioassay. . . . . . . . . . . . . . . . . . . 69 Growth Study. . . . . . . . . . . . . . . . . 71 Pond Studies . . . . . . . . . . . . . . . . . . . 72 ZOOplankton . . . . . . . . . . . . . . . . . 72 Benthos . . . . . . . . . . . . . . . . . . . 93 Fish. . . . . . . . . . . . . . . . . . . . . 99 Direct Toxicity. . . . . . . . . . . . . 99 Indirect Toxicity. . . . . . . . . . . . 99 Fish Growth. . . . . . . . . . . . . . . 100 Fish Reproduction. . . . . . . . . . . . 104 SUMMARY . . . . . . . . . . . . . . . . . . . . . . . . 107 PART III EFFECTIVENESS OF COPPER SULFATE AS AN AQUATIC HERBICIDE INTRODUCTION 0 O O O O O O O O O O O O O O O O O O C O O 109 MATERIALS AND METHODS . . . . . . . . . . . . . . . . . 111 RESULTS AND DISCUSSION. . . . . . . . . . . . . . . . . 112 SUMMRY O O O O C O O O .7 O O O O O O C C O C O O O O O l 2 2 REFERENCES CITED. 0 O O O O O O O I O I O O O O O O O O 123 APPENDIX. C O O O O I O O O O O O O O O O O O O O O O O 129 iv l “‘ . “mud B~6. E7. C‘l. COpper Copper Backgr Ratios dried centra sulfat Organi Bioass Weight Study. Change pond ( Changé FOUd ( pond ( Chang( Pond ( Chang' litEr TABLE LIST OF TABLES PART I Copper sulfate dosage rates and schedules. . . . Copper levels in natural systems . . . . . . . . Background c0pper levels from present study. . . Ratios of approximate peak copper values in dried plant tissue to theoretical initial con- centrations in water after dosage with copper su1fate. O O O O O O O O O O O O O O O O O I O 0 PART II Organisms common in outdoor study ponds. . . . . Bioassay results . . . . . . . . . . . . . . . . Weight changes in green sunfish during growth study. . . . . . . . . . . . . . . . . . . . . . Changes in numbers of benthic organisms/m2 in NW pond over time 0 O O O I I I I O I O O O O I O 0 Changes in numbers of benthic organisms/m2 pond Over time 0 O O O O O O O O O O O O O O O 0 Changes in numbers of benthic organisms/m2 pond over time . . . . . . . . . . . . . . . . . Changes in numbers pond over time . . . . . . . . . . . . . . . . . PART III Changes in numbers of NW pond phytoplankton/ liter over time 0 O O O I C O O O O O O C O C O O Page 18 19 38 68 70 71 94 95 96 97 113 :15? OF TABLES Changes liter c Changes liter < Change: liter < Per ce Reprod Green LIST OF TABLES--Continued TABLE C-2. Changes in liter‘over Changes in liter over Changes in liter over numbers time. . numbers time. numbers time. . of NE pond phytoplankton/ of SE pond phytoplankton/ APPENDIX of SW pond phytoplankton/ Per cent recovery of copper from spiked samples. Reproducibility of sample readings . Green sunfish stomach analyses . vi Page 114 115 116 130 131 132 IT “'9' H ..ch $10. A~11. $12, $13. Change in SW Change in NW Change in NE Change in SE Change Centre Change CEDtre Change centre Chang Centre COPpe: OVer . Coppe OVer COPPe Coppe addit COPPe ()‘7631: FIGURE A-lo o A-llo A-12. A-13. LIST OF FIGURES PART I Changes in water chemistry parameters over time in SW pond O O O O O O O O O I O O O O O O O O 0 Changes in water chemistry parameters over time in NW pond. O O O O O Q ~ 0 O O O C O O O O O O 0 Changes in water chemistry parameters over time in NE pond O O O C O O O C O O O O O O O C O O 0 Changes in water chemistry parameters over time in SE pond O O O O O O I O O O I O O O I O O O 0 Changes in dissolved and suspended copper centrations over time in SW pond. . . . . . . . Changes in dissolved and suspended c0pper centrations over time in NW pond. . . . . . . . Changes in dissolved and suspended copper centrations over time in NE pond. . . . . . . . Changes in dissolved and suspended copper con- centrations over time in SE pond. . . . . . . . Copper accumulation in NW and SW pond sediments over time 0 O O O O O O O I O I O '0 O O O O O O COpper accumulation in NE and SE pond sediments over time 0 O I O I O O I O O O O O O O O O O 0 Copper accumulated by plant tissues in SW pond ove r time 0 C O O O I O O I O O O I O O O C O 0 Copper accumulated by plants growing after addition of copper sulfate to SW pond . . . . . Copper accumulated by plant tissues in NW pond over time . . . . . . . . . . . . . . . . . . . vii Page 11 13 21 25 28 30 33 36 40 43 45 LBTOF FIGURES .ofly' " :N'LRL A44.Copper a over tin - . Copper a NW ponds - . Copper 5 SE ponds - . Copper 5 over tin - . Per cent ‘1’ ' __4 KO 0 11. $3. $4. $5. E~6. $7. various age unti Per cent various age unti Changes time, . . Changes Species Changes time. . Changes species Changes time, . Changes Species Changes LIST OF FIGURES--Continued FIGURE A-l4. A-15. A-16. A-17. A-180 A-lgo Page Copper accumulated by plant tissues in NE pond over time I I I I I I I I I I I I I I I I I I I 4 8 COpper accumulated by green sunfish in SW and NW ponds over time. . . . . . . . . . . . . . . 51 Copper accumulated by green sunfish in NE and SE ponds over time. . . . . . . . . . . . . . . 54 Copper accumulated by snails in NE and SE ponds over time . . . . . . . . . . . . . . . . . . . 57 Per cent of total applied copper present in various components of SW pond from time of dos- age until end of experiment . . . . . . . . . . 60 Per cent of total applied copper present in various components of NW pond from time of dos- age until end of experiment . . . . . . . . . . 62 PART II Changes in SW pond zooplankton numbers over time I I I I I I I I I I I I I I I I I I I I I I 74 Changes in numbers of SW pond cladoceran species over time . . . . . . . . . . . . . . . 77 Changes in NW pond zooplankton numbers over time I I I I I I I I I I I I I I I I I I I I I I 79 Changes in numbers of NW pond cladoceran species over time . . . . . . . . . . . . . . . 82 Changes in NE pond zooplankton numbers over time I I I I I I I I I I I I I I I I I I I I I I 85 Changes in numbers of NE pond cladoceran species over time . . . . . . . . . . . . . . . 86 Changes in SE pond zooplankton numbers over time I I I I I I I I I I I I I I I I I I I I I I 88 viii 1:510? FIGURES IIHVV 0 ‘ .sdva FL Cl. Changes species Mean pe: fish in . Mean pe. fish in Changes NE: and LIST OF FIGURES--Continued FIGURE Page B-8. Changes in numbers of SE pond cladoceran Species over time . . . . . . . . . . . . . . . 9l B-9. Mean per cent weight increases for green sun- fish in NW and SW ponds over time . . . . . . . 102 8-10. Mean per cent weight increases for green sun- fish in SE and NE ponds over time . . . . . . . 106 PART III C-l. Changes in P04 concentrations over time in NW, NE ' and SE ponds I I I I I I I I I I I I I I I I 119 ix The effe Various doses Pieter, sedime lyzed for co; Liportant ro] Effects and green sur :cnducted on (A) fCOPPer 110) Of 200;)15mkt< 0f COpper 0n 'i'ere ana1y2e< Effecti‘ in the Prese; lacrophyteS tem‘lned a ft In Part effeCts of C m in Part Stu . dy ls ana INTRODUCTION The effects and fate of copper in ponds were studied. Various doses of copper sulfate were added to two ponds. Water, sediments, plant tissues, fish, and snails were ana- lyzed for c0pper content to determine which components played important roles in copper dispersal. Effects of copper on populations of zooplankton, benthos, and green sunfish were studied. Laboratory bioassays were conducted on pond organisms to determine approximate levels of copper toxicity. In the pond study, changes in populations of zooplankton and benthic organisms were determined. Effects of copper on green sunfish survival, growth, and reproduction were analyzed. Effectiveness of copper sulfate in algae and weed control in the present study was noted. Changes in populations of macrophytes and filamentous and planktonic algae were de- termined after copper sulfate applications. In Part I, fate of copper is discussed; in Part II, effects of copper on fish and their food organisms are noted; and in Part III, effectiveness of c0pper sulfate in the present study is analyzed. PART I FATE OF COPPER IN PONDS Kopp and mr natural activities in industrial ef L) f mdesirabl Relative LI 0 : Copper in {1969), and C hehavior in c ville (1970). inveStivatea IESpectiVEly the impact 0 Persa1_ Man alkaline Wa t tance of Wat dispers a1 ir INTRODUCTION Kopp and Kroner (1967) note that low levels of copper occur naturally in water. Sources of copper from human activities include leakage from copper and brass tubing, industrial effluents, and use of copper compounds for control of undesirable aquatic organisms or plants. Relatively few studies have been conducted on the fate of copper in aquatic systems. Bartley (1969), Nelson gt 31. (1969), and Chancellor gt 31. (1958) have studied copper behavior in canals and irrigation ditches. Deubert and Demoran- ville (1970), Riemer and Toth (1968,1970), and Riley (1939) investigated c0pper dynamics in bogs, ponds, and lakes, respectively. Little work, however, has been done to estimate the impact of the various components of ponds on copper dis— persal. Many early workers felt that copper, when added to alkaline waters, precipitated rapidly as a basic, insoluble carbonate. This paper describes an attempt to determine the impor- tance of water, sediment, plant tissue, and fish in copper dispersal in ponds. Fhvsical Des: The two study are loc Plant, now us Fisheries anc‘ Ear ponds, 1c and 2 m deep. In June CO‘Jered with mentous a1 gas aquatic inSec In earl) ene Sheet, El shallow grave aeratEd glas c MATERIALS AND METHODS Physical Description of Ponds The two ponds (old trickling filters) in the present study are located at the old East Lansing Sewage Treatment Plant, now used by Michigan State University's Department of Fisheries and Wildlife for limnological research. The circu- lar ponds, located side by side, are about 13.7 m in diameter and 2 m deep. In June 1970, the ponds were drained, and the bottom was covered with 10 to 15 cm of black soil. The ponds were re- filled from an adjacent pond and left undisturbed until Spring 1971. During summer 1970 pOpulations of macrophytes and fila- mentous algae had developed; large numbers of zooplankton and aquatic insects were also evident. In early May 1971 each pond was divided with a polyethyl- ene sheet, effectively sealing off two systems. In early April green sunfish (Lepomis cyanellus) were collected from a shallow gravel pit near Williamston, Michigan, and held in aerated glass tanks. In June 100 fish were weighed, marked by fin-clipping, and released into each pond. Copper sulfate solutions were prepared in distilled water from commercial grade copper sulfate pentahydrate (CuSO .5 H20) and broadcast 4 by hand over the ponds according to the schedule in Table A—1. Table A-1 - ( # —————— Pond Scuth half 01 (SW) -tre Earth half 01 (NW) -cox Earth half 01 (NE) -cox Sauth half 01 (5E) ~tre Control later in the iiSperSal Of 3360 throughc an: ' “roxlmatell mi . Mammal Desc Watel. ct alkalinitv. r det . mined tt Table A-1. Copper sulfate dosage rates and schedules. Pond Dosage Dosage Date South half of west pond (SW)-treatment July 12 3 ppm copper sulfate North half of west pond (NW)-control Sept. 26 3 ppm copper sulfate North half of east pond (NE)-control Oct. 18 1 ppm copper sulfate South half of east pond (SE)-treatment Aug. 16 1 ppm c0pper sulfate Sept. 16 1 ppm c0pper sulfate Oct. 7 3 ppm c0pper sulfate Nov. 4 5 ppm copper sulfate Control ponds (northwest and northeast) were treated later in the study to obtain further data about effects and dispersal of copper. The abbreviations SW, NW, NE, and SE are used throughout the three parts of this paper to refer to the various ponds. Note that 1 ppm copper sulfate represents approximately 0.25 ppm copper. Chemical Description of Ponds Water chemistry parameters, including temperature, pH, alkalinity, hardness, and dissolved oxygen (D.O.) were determined throughout the study period (Figures A-l through A-4); phosphorus as P0 was measured in three of the systems 4 late in the study (Figure C-l). Methods outlined in Standard Methods for the Examination of Water and Waste Water (1965) Figure A-l. Changes in water chemistry parameters over time in SW pond. Each point represents the average of two samples taken from the top (solid line) or bottom (broken line) of the pond. Shaded area denotes addition of 3 ppm copper sulfate. 90- 60! at 20+- Ir?” 9.0 1 3.0 130 ‘ leg .. 140 i Lag 6~11 Phenolphthalein Alkalinity (As ppm CaCOB) Methyl Orange Alkalinity (As ppm CaCOB) Dissolved Oxygen (ppm) pH 180 TI, Hardness (As m CaCO ) 160 “ ..... pp 3 140 120 l l 1 4 1 J~__ 6-11 10-9 Figure A-l Figure A-2. Changes in water chemistry parameters over time in NW pond. Each point represents the average of two samples taken from the top (solid line) or bottom (broken line) of the pond. Shaded area denotes addition of 3 ppm copper sulfate. 150_ 120} 9.0- 6.0~ 10* 10.0" 3.0 7,0 130 160 140 12:) 80 Phenophthalein Alkalinity (As ppm CaCO3) 60 40 20 160 Methyl Orange Alkalinity (As ppm CaC03) 120 80 Dissolved Oxygen (pew pH Saw Hardness ‘"” (As ppm CaCOB) Figure A-2 Figure A—3. 10 Changes in water chemistry parameters over time in NE pond. Each point represents the average of two samples taken from the top (solid line) or bottom (broken line) of the pond. Shaded area denotes addition of 1 ppm copper sulfate. 1w~ 1m“; awx 40‘ 12.0 )— 9.0 +- 6.0 " 10.0 t‘ to‘ 3.0 80 60 40 20 160 120 80 40 12.0 9.0 600 3.0 10.0 9.0 8.0 150 130 110 90 11. - .yfi; Phenolphthalein as ADmlhflty Eff? (As ppm CaCO3) ..... .. .a ...... ...... ..... ...... ..... ...... ..... ...... ...... ..... ...... ..... ..... ...... ..... ...... ..... ..... ...... ..... ...... ..... ...... ...... ..... ...... ...... ..... ...... ..... ...... ..... ..... ,sfl Methvl Orange gee Alkalinity ..... ..... gflfig (As ppm CaCUJ) 32.55:} DiSSOlved Oxygen I... P........goooo.ooooo ...... ...... . ...... ..... P ,,,,,, . ..... ..... ...... ...... ..... ...... ..... ...... ..... ..... ..... ...... ...... ..... ...... ..... ...... ...... ..... ...... ..... ...... ...... ” ggfi; Hardness iflfii (As ppm CaCO3) -‘.u l I'Z'I‘I'I-I 1 L 6-11 7-1 7-21 8-10 8-30 9-19 10-9 10-29 11-18 Figure A-3 Figure A-4. 12 Changes in water chemistry parameters over time in SE pond. Each point represents the average of two samples taken from the top (solid line) or bottom (broken line) of the pond. Shaded areas denote additions of 1, 1, 3, and 5 ppm copper sulfate. 80—- 40 20 0- 1mm l'. 1m' 20.0 _ Unfi in- 10.0 > 9-01)‘ 8n, 7.0 r 180 ‘ 1 160 140 120 13 80 60 40 20 160 120 80 40 20.0 §§§§ LE? ?E¥ 2 i oxygen mu :2: igflpmw . . ..... 180 160 140 120 Figure A-4 were used fl analysis we with a Kemm Sampling ar Water near the c. account fo rents. Wa “as collec through a while tha‘ Suspended 14 were used for chemical tests. Replicate water samples for analysis were collected from the top and bottom of each pond with a Kemmerer water sampler. Sampling and Sample Preparation Vla_te_£ Water samples were taken at three stations established near the center, middle, and edge of each semicircular pond to account for differences in copper concentrations due to cur— rents. Water was removed with a core sampler, and a subsample was collected. Preparation followed procedures in Methods for Chemical Analysis of Water and Wastes (1971). C0pper passing through a 0.45-u membrane filter represented dissolved copper, while that fraction remaining on the filter was considered suspended c0pper. Sediments Two sediment samples were collected with a core sampler each sampling period, frozen in dry ice, and later dried at 100 C for 96 hours. Samples were ground, and 5 m1 of 8 N HCl per gram of sediment were added. This slurry was heated near boiling for 20 hours, cooled, filtered, and diluted to volume with deionized distilled water. Plant Tissue Replicate samples of plant tissues were collected, rinsed in distilled water, and, after blotting, dried at 100 C for 96hours. and boiling mixture reh filtered an Fish v and digeste M hours. Snails dried. The CODDer w COPpe tiOn Spect. de+ Q sulfate 8 t inSUre ace by a diffe Durin with Coppe 3 15 96 hours. Tissues were digested by adding 10 ml of 8 N HNO3 and boiling to dryness; 5 ml of 6 N HCl were added and the mixture reheated to dissolve the residue. The solution was filtered and diluted to volume. Fish Fish were trapped, rinsed in distilled water, weighed, and digested in 5 ml of concentrated HNO per gram of fish for 3 24 hours. The solution was filtered and diluted to volume. Snails Snails were collected, rinsed in distilled water, and dried. They were weighed, digested in 10 ml of concentrated HNO to dryness, and then redissolved in 5 m1 of 6 N HCl. 3 Filtering and sample dilution followed. Copper Analysis Copper was determined with a Jarrell-Ash Atomic Absorp— tion Spectrophotometer Model 800. Sample concentrations were determined by comparing their readings to those of copper sulfate standards made up several times during the tests. To insure accuracy, three standards were compared to some made up by a different investigator; the discrepancy was less than 10%. During the analyses samples from control ponds were spiked with copper to determine per cent recovery (Table D-l, Appendix). Recoveries from plant tissues ranged from 89.9% to 114.4% with azean of 1 from 94.7% Reproc samples on gairs of re significani Coppe: Little dowr COPPGI in ‘ he“? Core iCCumulati< 16 a mean of 101.2%, while recoveries from sediments ranged from 94.7% to 120.0% with a mean of 109.9%. Reproducibility was determined by reading the same samples on different occasions (Table D-2, Appendix). The pairs of readings, when tested with a paired t-test, were not significantly different at the 5% level. Copper added to ponds accumulated on sediment surfaces. Little downward movement of c0pper was expected. Expressing copper in terms of sediment weight was deemed meaningless, as a heavy core sample, while presenting the same surface area for accumulation as a smaller one, would have a lower c0pper con- centration based on weight. Copper concentration based on surface area was determined as follows: (1) a background value in mg copper/kg of sedi- ment was calculated for untreated sediment samples; (2) this concentration was subtracted from total copper determined for each treated sediment sample; and (3) the remaining copper was assumed to be that accumulated on the sediment surface during the study. Backzround Table systems; t occurring levels in generally Because ex vary widel o. RESULTS AND DISCUSS ION Background Levels Table A-2 lists natural copper levels found in other systems; these levels are assumed to represent only naturally occurring copper with no introduced copper present. Copper levels in components from the present study (Table A-3) generally agree with levels found in past work (Table A-2). Because experimental techniques and recovery efficiencies vary widely among workers, values listed represent general estimates. Dynamics of Copper Added to Ponds m SW Pond Dissolved c0pper dispersal in SW pond exhibited two phases: a rapid decline following initial peaks and a subsequent gradual decrease over time (Figure A-5). The large peaks noted at stations SW1 and SW2 shortly after dosage were probably due to slow dispersal of copper sulfate. The likely cause of the rapid decline was a precipi- tation, probably of malachite (Cu2(OH)2C03). Stiff (1971) noted that in waters with alkalinity and pH similar to those in the present study, malachite, rather than cupric hydroxide l7 Noo.o HTHMB UCOQ .Hm 00 C0 AEQQV HQQQOU 000H0>4 GONZO“ MHOUUmflUWQ>CH . UGQCOQEOU uhflflhflvumxm .Huu..H-...UHWFN F~fi. mafiflvkrflvifi Hallou nNInA‘ «WNQIWPNL. 18 pououaflm: “momma Hmuaflm NH .02 Queues: smsounu poumuawmm “momma uwuaflw vv .oz Imusmmmum onwm whom :Im.o nmsounu commuaflmu .mwsum wuoumuonma m ca poms swam Eonm mam>mqo unaw>ma moon 36:3h “mamas unmfloz whom “mammn unmflm3 amen “Hmuawm :Imw nmsounu “Houaflm mmmamwxmam oaMHQEoE unaumsz nwsounu omumuaflm. mm.a .Hw MW .mwflumm usouu 3oncwmm .vm.a .HmlyWI.msflume gmemcaoo on.m :mmfiSOHz oxmq .Hm MW .mmosq noummusoue om.o :mmw30Hz oxen .Hm.ww .mmosq Hwnwnm pouuomm mm.o :mmHQOHz oxen .Hm um .mmosq mma3mam bmm: m.mm muoumz wmmumb 3oz such .umemflm .mm dunno m.>H Hmsmo GOwummfiHHH moapumm msmowaow soummoemuom m.mv mumum3 ammumw 3oz suoe .HmEmHm msmwfluo coummoemuom m.a~ Amado coflummHHHH weapumm mflmcopmcmo mmpoam m.am muoum3 mmmumh 3oz nuoe .HmEmHm .mm mmUon .mpmmHe ezflu .m .D Honoux .mmox nmmo.o mnmumz pcom Impmm .HmEmHm Hoo.o «Adamo coflummHHHH .Hm um .comamz mo.o dMounts mop wuumncmuo mHHH> IGMHOEmQ .uumnsmo mvo.o Amado nowummflHHH .II II mmauumm moo.o Hmum3 pcom .Hm um .couusm mma¢3 Afimm% Hommoo mmmuo>¢ monsom muoummwumw>cH ucwcomfiou .mfioumMm Humane: :H mao>ma Hmmmou .Nua mfinma 'I“ III-I '- 'l.‘ MO HQQESZ Occmm .E:C\ x. “ n a my H E l9 .mflmon unmflos meow umfimon usmwoz uo3lmpon oHoszm umflmon unmflos mun: umwmon unmfloz muam “Houaflm ocoHQEoE so umoa coauooumu “Houaflm osounfioa numv nmsouau UoHouHHmH o Hm.~meu om.mn mm.mm .dm mmxsm mqu¢Zm mm vw.a oo.o mmwm.o nuancSm soonw mmmHm om om.mm mm.v Hm.mm mommfiuo :ouomofiouom mm Hm.om mm.m Hm.mm Hflaaouusz oopoam ma mm.Hm vm.mH oe.qm .dm guacamoemo om oe.om mo.m mm.ma .dm mumno smsmmHB Bzmqm Hm om.mn mo.H mm.hm mmezmzHomm mm aoaa.o Imoo.o Nomo.o Houoa mm momo.o oo.o hmmo.o Npopsommsm mm mo.o oo.o meao.o spo>H0mmHQ mmsdz moamfiom mo HoQEsz omcom Afimmv Hommoo omouo>¢ unocomeou .hpsum ucomonm Scum mHo>oH Hommoo pcsoumxoom .mIG oHQoB iFigure A-S. 20 Changes in dissolved (solid line) and suspended (broken line) copper concentrations over time in SW pond. Shaded area denotes addition of 3 ppm copper sulfate. MG COPPER/LI TEN 1.0 0.6 A .° ijWTTT'II I o N T 0.25 6~25 MG COPPER/LITER 10° - 21 ..... ..... ..... ..... ..... ..... ..... ..... ..... ..... ..... ..... ..... ..... ..... ..... E; '33- O ...... .llcoloooo.ouooo‘ucoco ‘. ..... ..... ..... ..... ocean-0.0.... l 0,2.— 6-25 . .. r . .. . . .. . . .x r. .. . . .. .. . - a. -. . . .. .. ., . .. -. . . . .. .. . .. .. .. . . .. .. .. .. . ,. .. . .. .. . .. .. .. . . .. .. .. .. . . .. .. . . . . '.' .. -. .. . . -. .. . . -. . . . .. . . .. .. - . ,. .. . .. o . . .. - V . .. . . . .. . . . .. . . ... . . n... . . -\\huufluu%.u .. .oooIo-I.0e0nvu.u. noon-non . n o ..I oIoI-nIIIounuuocounts-03.0.0.0.I I I I I I I I I o I I a or... I.I I.IIoIc II: Icon-I. eI- n nIIIoI- ~II I p.- o .I. M: . . .. . 'A -.I ounuIIIcII IIIInIIIusn - n I u I. III-a I 10-4 m. 0 1 9-14 ..I00. ......IIIII 00... 8-25 TIME Figure A-8 only 20% (1970) f. 14% afte sulfate. three No after do and inso A r studies that COp Organic 31 only 20% and 36% of the total copper applied. Sutton SE.E$° (1970) found 36% of applied copper remaining after 7 days and 14% after 21 days following dosage of small pools with copper sulfate. Smith (1939), however, found that copper levels in three Nova Scotian lakes with acidic water remained high long after dosage; he attributed gradual copper losses to drainage and insoluble complex formation with organic matter. A rapid loss of dissolved copper has occurred in most studies done in alkaline water. The present study indicates that c0pper may remain in solution longer than expected; organic complexes are a possible mechanism for this phenomenon. Sediments SW Pond Upon dosage, sediment accumulated copper rapidly at first and gradually later (Figure A-9). The mechanism involved may have been the following: there was an initial precipitation of malachite and insoluble organic complexes; direct adsorp- tion of ionic copper also occurred. An equilibrium was slowly reached between sediment copper and complexed forms in water, resulting in a gradual accumulation. Colloidal suspensions of copper with various organic components gradually precipitated and added to copper concentrations. NW Pond Copper accumulation in NW pond sediments occurred more quickly than in those of SW pond, where a delay of several days 32 10000 _ 1000 r— H O O I Figure A-9. Copper accumulation in NW and SW pond sediments over time. Shaded areas denote addition of 3 ppm copper sulfate. H O I 10000 1000 (.16 COPPER/UNIT SURFACE Am or BEDININ'I' Mo. 10000 #— 1000 ~— 100 - 10000 H O O O 06 COPPER/UNIT SUREACE ARIA OP SEDINBNT 100 10 H O I 33 Northwest Pond .... s ... .... II. .... u'I.-'u A... l .... -.-. ...A 7-21 Figure A-9 Southwest Pond l .1_- 10-29 .-__-..- ..,I 11-18 existed Otherwis similar. Pos slightly so: pond occurred Sim As ual than 0f accum tates in imPOI‘tan percenta 34 existed before elevated levels of copper appeared (Figure A-9). Otherwise, the accumulation pattern in the two ponds was similar. NE Pond Post-treatment analysis of NE pond sediment showed only slightly elevated copper concentrations (Figure A-lO). As NE pond was treated with only 0.25 ppm copper and dosage occurred late in the season, these results were not surprising. SE Pond As copper accumulation in SE pond sediment was more grad- ual than in other ponds (Figure A-lO), a different mechanism of accumulation may have been operative. The role of precipi- tates in initial sediment accumulation may have been less important if the bacteria present accumulated significant percentages of the applied copper. Involved in sediment copper accumulation were not only precipitates, such as malachite, and insoluble organic com- plexes, but also ionic or complexed copper adsorbed directly from water. Tobia and Hanna (1958) stated that copper reten- tion in soils was correlated mainly with amount of organic matter in the soil. Lucas (1948) noted that organic soils held copper tenaciously in the zone of placement. Riemer and Toth (1970) concluded that copper was rapidly adsorbed by pond sediments and that amount of organic matter and nature Figure A-lO. 35 Copper accumulation in NE and SE pond sedi- ments over time. Shaded areas denote additions of 1 ppm copper sulfate in NE pond and of 1, SE pond. 1. 3, and 5 ppm c0pper sulfate in 10000 F 1000 fit H O O I H 0 V1 10000 .. UG COPPER/UNIT SURFACE AREA 0!" SEDIMENT H O O O 7 ' moL 10000 1000 100 H 0 10000 UG COPPER/UNIT SURFACE AREA OF SEDIHENT p... O O O 100 10 136 Southeast Pond I Northeast Pond 1 :ééiiL 1 1 9-30 10-20 11-9 11—29 TIME Figure A-10 J 12-19 of clay mi fixed. As se matter (ll reached th reducing c CuS at the significan retention Accum shown by O averaging after trea (1969) fou canal afte 99 days du neXt YEar, AVail mate rePOs COPPEr Com PIEXeS fro Results fr baCteria, initial bl. 37 of clay minerals present determined the amount of copper fixed. As sediment in the current study was high in organic matter (11%), copper probably remained on the bottom once it reached the sediments. Hutchinson (1957) noted that under reducing conditions, copper remained largely immobilized as CuS at the mud surface. Even during oxidizing conditions, a significant copper release should not occur since copper retention is high in organic soils. Accumulation of introduced copper in sediments has been shown by other workers. Nichols EE.El° (1946) found levels averaging 420 mg copper/kg dry mud for Lake Monona, Wisconsin, after treatment with a total of 170,000 kg of c0pper. Bartley (1969) found levels of up to 352 mg copper/kg dry mud in a canal after providing a constant flow of 0.17 ppm copper for 99 days during one year and 0.11 ppm copper for 70 days the next year. Available evidence indicates that sediment is the ulti- mate repository of c0pper added to a pond. Precipitation of copper compounds followed by gradual adsorption of copper com- plexes from water probably occurred in the present study. Results from SE pond indicated that large populations of bacteria, and perhaps algae (Toth and Riemer, 1968), may delay initial buildup of copper in sediment. m Plant levels (Fi the theore for the th Table A-4. Plant tiss after dose 38 Plant Tissue SW Pond Plant tissue rapidly accumulated copper to very high levels (Figure A—ll). The ratio of copper in dried tissue to the theoretical initial concentration in water was above 3,000 for the three species present (Table A-4). Table A—4. Ratios of approximate peak copper values in dried plant tissue to theoretical initial concentrations in water after dosage with copper sulfate. Plant Pond SW NW NE ghaga sp. -- 1360 2649 Potamogeton crispus 4616 1716 3426 Elodea Nuttallii 3018 3329 1832 Oedogonium sp. 4358 2846 -— The rate of accumulation for Elodea Nuttallii was lower than for either Oedogonium sp. or Potamogeton crispus. The declining copper concentrations in Elodea (Figure A-ll) are indicative of tissue destruction in the plant after c0pper application. Loss of leaf tissue and destruction of root attachment was nearly complete; copper decrease in these plants was thus probably due to leakage. Copper concentrations in macrophytes which developed in SW pond after treatment were slightly above background levels Figure A-ll. 39 Copper accumulated by plant tissues in SW pond over time. Shaded area denotes addition of 3 ppm copper sulfate. MG COPPER/KG PW? TISSUE (DRY NT) 5000 1000 100 L tifldskfidaog,fi-V H 'J‘.;' at! 7~I MG COPPER/KG PLANT TISSUE (DRY WT) 5000 1000¢ 100 ..... ..... ..... ..... 40 Figure A-ll o Oedogonium sp. a Potamogeton crispus I Elodea Nuttallii (Figure A- ring. CO] ably not : Bartley (I contained weight am the hydro: Coppe pond (Fig and Were 1 The 5 c lc resPOns ib; COppE P WE 900d, Whex two Specie actiVitieE dOSage takl notEd tha be relate. growth ch detemline The COpPEr 1e Stipend. 41 (Figure A-12); copper uptake from sediment was possibly occur— ring. Copper levels in water were near background and prob- ably not responsible for the elevated concentrations noted. Bartley (1969) found that pond weeds in an irrigation canal contained copper levels of from 36 to 143 mg copper/kg dry weight and stated that accumulation of residual c0pper from the hydrosoil occurred. NW Pond Copper was taken up rapidly by the plants present in NW pond (Figure A-l3). Levels were generally lower (Table A-4) and were reached more slowly than in plants present in SW pond. The 5 C lower water temperatures in NW pond may have been responsible. Copper levels in Elodea were higher than those in either Potamogeton or Oedogonium; this pattern contrasted with SW pond, where Elodea levels were lower than those in the other two species. Thus uptake may have been related to growth activities, with those species growing most rapidly during dosage taking up the most c0pper. Sutton and Blackburn (1971) noted that susceptibility or resistance to copper sulfate may be related to the manner in which c0pper is taken up by plants; growth characteristics of each species at dosage time may determine the degree to which each is affected. The only plant surviving treatment, Elodea, retained copper levels deSpite sustaining damage similar to Elodea in SW pond. Figure A—12. 42 Copper accumulated by plants growing after addition of copper sulfate to SW pond. m COPPER/KG PM"? TISSUE (DRY HT) 5 H u C 90 50 10 MG COPPER/HG PLANT TISSUE (DRY HT) 110 130 90 50 10 43 a Potamogpton crispus _' . aggrilla verticillata . Elodea Nuttallii I h.- I P I I r' I 0 E O h o 2 I I . D l— O I l— L L l 9'15 10'5 10-25 11-14 TIME Figure A-12 Figure A-l3. 44 Copper accumulated by plant tissues in NW pond over time. Shaded area denotes addi- tion of 3 ppm copper sulfate. MG COPPER/KG PLANT TISSUE (DRY HT) 5000 - 1000 I 100 h. 50 MG COPPER/KG PLANT TISSUE (DRY NT) 45 5000 [— A Chara sp. a Potamogeton crispus o Oedogonium sp. I Elodea Nuttallii 1000 — § I 100 —’ ..... a o I 501__iP o z o 0 o o O 8 . D I o O A 69 o g 3 a I ' 0 .3 CDO I 0 I ' '9 I I f" ii _ 1 J 1 '1" 7-8 7-28 8—17 9-6 9-26 TIME Figure A-13 Macru (Figure A; applied , Uptake ra however . ment reta suffered Slowly fr Concentra the mac rc Sutt Phl’te m ism, aCm This Pher Upta thOSe fr( 673 ppm ( after trv 161th 0v. original 46 NE Pond Macrophytes in NE pond showed a rapid uptake of copper (Figure A-14). Since only one-third as much copper was applied, levels were lower than in either SW or NW ponds. Uptake ratios (Table A-4) were as high as those in NW pond, however. Levels were attained more slowly than in SW pond. No important differences in species uptake were noted. QEEEE and Elodea samples taken at the end of the experi- ment retained high copper concentrations. Since these species suffered little injury, copper was apparently released very slowly from undamaged tissues. A natural decrease in copper concentrations on a per gram plant tissue basis occurred as the macrophytes grew. Sutton and Blackburn (1971) noted that the aquatic macro- phyte Hydrilla verticillata possessed an active uptake mechan- ism, accumulating copper against a concentration gradient. This phenomenon occurred in the current study. Uptake rates and levels in this study were similar to those from past work. Sutton and Blackburn (1971) treated Hydrilla with 0.25 ppm copper (as copper sulfate) and found 673 ppm copper in the tissue on a dry weight basis 4 days after treatment. Plants treated with 32 ppm copper accumu- lated over 30,000 ppm. The ratio of copper in plants to original copper in ambient solutions declined at concentrations greater than 2 ppm; injury to plant tissues probably caused Figure A—l4. 47 Copper accumulated by plant tissues in NE pond over time. Shaded area denotes addi- tion of 1 ppm copper sulfate. MG COPPER/KG PW? TISSUE (DRY HT) 5000 1000 *- 201 no COPPER/KG PLANT rrssuz (on! NT) 5000 1000 100 20 _ 48 .... ..... -... ..... ....... ..... . Elodea Nuttallii ea ..... .... ----- ..... ..... ‘ Chara .po 1:55:35 ..... ....... ..... ..... a Potamogeton crispus :mw ..... ..... ..... . Spirogyra sp. g; - a ..... ..... ..... ..... ...... aaaaa ...... ..... .... ..... ----- e ...... ..... ..., ....... ..... ...... ..... ..... ....... .... s. . ..... ..... ..... '''''''' ID 9 Figure A-l4 11-6 11-26 I I}... 12-15 this decli in the fir Suttc with 1 ppm Eggs, 132 dynamics i and three water cont to 4820 or in this st trating Cc Diffe grosvth cor warm Wate: SpeCies di PhYSiolog: AS illLISt] macrophl/te NW and \Sw In bc showed 1m again afte backg r Ouné 49 this decline. The authors noted that copper uptake was rapid in the first 2 days and gradual thereafter. Sutton 23 a1. (1970) placed Hydrilla verticillata and Najas gpadalupensis cuttings in outdoor pools and treated them with 1 ppm copper. Hydrilla accumulated 2996 ppm copper and Najas, 1325 ppm, after 3 days. Bartley (1969), studying copper dynamics in an irrigation ditch. noted that Elodea canadensis and three species of Potamogeton concentrated copper from water containing about 0.1 ppm copper by factors of from 340 to 4820 on a wet weight basis. As literature values and those in this study indicate, aquatic plants are capable of concen- trating copper to very high levels. Differences in uptake between species seemed related to growth conditions of plants, with an actively growing crop in warm water having the greatest uptake. When determining species differences in copper toxicity to plants, however, physiological reactions must be considered in greater detail.‘ As illustrated in SW pond, elevated copper levels in new macrophytes seemed due to c0pper uptake from sediment. Fish NW and SW Ponds In both ponds slight increases in fish copper concentra- tions after treatment occurred (Figure A-15). Fish in SW pond showed levels of about 3 ppm copper 1 week after treatment and again after 47 days. By the 79th day, levels decreased to background values of about 0.5 ppm copper. Figure A-lS. 50 Copper accumulated by green sunfish in SW and NW ponds over time. Shaded areas denote addition of 3 ppm copper sulfate. WHOLE FISH (WET WT) MG COPPER/KG soL 4.0!. 2.0}- 1.0,» 51 l . J .m I IIIII .m 0 0 II I II. P P t t s s w. w h h t t w m o S N l. I I IIIIIII III _ I I I II I m I III I . .IL IIII b w I I IL II _ _ l a I I _ _ III II I I I 1* p _ _ r H o o O I I I 3 2 1 ~83 Bfllv :mHh WAGE: 0&\mflmm00 0! 10-25 11-14 10-5 8-12 7-20 TIME Figure A-lS Fish 10 days a occurred Fist after dos dosage ir trations Fisl anaverac i“ a tOte were ant: 52 Fish in NW pond showed copper increases to about 2 ppm 10 days after treatment; decrease to background levels occurred by the Slst day. NE Pond Fish in NE pond showed no increased copper concentrations after dosage (Figure A-l6). Since the level of copper sulfate dosage in NE pond was low, the lack of elevated copper concen- trations in NE pond fish was not surprising. SE Pond Fish collected from SE pond at the end of the study had an average concentration of about 15.5 ppm copper (Figure A-16). As a total of 2.5 ppm c0pper was added to the pond, high values were anticipated. Fish accumulated copper from water but only to low concen- trations. Kariya 23 El- (1967) found an average of 1.34 ppm copper in control goldfish (whole body), while Specimens placed in 1 ppm copper solutions had an average of 6.43 ppm copper. Fish tested in 0.5 ppm copper accumulated copper to levels averaging 5.17 ppm. Copper accumulated in the mucus of gills of dead fish may have contributed to the levels noted in this study. Copper concentrations in green sunfish decreased quickly after copper sulfate treatment. Only in SE pond where dosages were multiple were levels above background at the end of the 53 Figure A-l6. Copper accumulated by green sunfish in NE and SE ponds over time. Shaded areas denote addition of 1 ppm copper sulfate in NE pond and 1, l, 3, and 5 ppm copper sulfate in SE pond. MG COPPER/KG WHOLE FISH (WET WT) 604>L 55.0 — 40.0" 35.0 v— 25.0 .— p U! I O I F o T N b 1 7-10 Southeast Pond Northeast Pond 8-2 8-22 54 ...... . . , ..... ooooo .u. ..... 10-15 TIME 11-4 Figure A-16 s.- all ..o- ... III. u... I... u... n..- no.- I-Io .nou non. .II. - v”q~--- u“ ......_..._ . . ---'- .. .— ‘I . ._ ..- . -... .—-—.--.—.a——c——-—I—- ___L_ --..____.1_._. 11-24 12-14 55 experiment. When copper is applied infrequently and at low levels to ponds, its accumulation in fish is probably not significant, as concentrations, although elevated temporarily, quickly return to normal. Snails NE and SE Ponds Snails accumulated high levels of copper (Figure A—l7). Higher values in SE pond than in NE pond may be attributed to higher water temperatures in SE pond during dosage. A second dose of 1 ppm copper sulfate applied to SE pond increased c0pper levels to about 850 ppm copper on a dry weight basis, approximately twice levels noted after the first dosage. Components of Copper Distribution Estimates were made of the fraction of applied copper found in each major component of NW and SW ponds. Plant values were calculated by estimating total weight of plant masses and total copper accumulated in the masses. Water concentrations included c0pper in dissolved and suspended states. Total sedi— ment concentrations were calculated by multiplying average copper values.derived for individual core samples (see methods section for this technique) by a factor derived by dividing total sediment area by core area. SW Pond Only water and sediment copper are graphed, as plant tis- sues contained less than 1% of total applied copper at any 56 Figure A-l7. Copper accumulated by snails in NE and SE ponds over time. Shaded areas denote addi- tion of 1 ppm copper sulfate in NE pond and two doses of 1 ppm copper sulfate in SE pond. HG COPPER/KG SNAIL TISSUE (DRY WT) 57 Southeast Pond Northeast Pond O 100_. 3 O o _______J_i_____1___.l_,1 ' 1 1 1 e-1o a—ao 9-19 1o-s 10-25 11-14 12-4 12-24 TIME Figure A—l7 58 time (Figure A-18). A clear relation existed between water and sediment COpper in SW pond: a rapid initial decrease in copper in solution with a concomitant increase in sediment copper values occurred. Copper in sediment at the end of the experiment was greater than 90% of the total applied. NW Pond Plant tissue, mostly Chara, played a significant role in NW pond c0pper dispersal (Figure A-19). Percentage of copper in plant tissues reached about 10% of the total applied on the 14th day. Copper was released shortly thereafter during decomposition. Sediment accumulation followed a pattern similar to that in SW pond. Copper values in water were influenced by release of copper from dying Chara tissues. 59 Figure A-l8. Per cent of total applied copper present in various components of SW pond from time of dosage until end of experiment. PER CENT OF TOTAL COPPER APPLIED 100 60 Om". SW Water o—n SW Sediment 0‘00.... '. l L 1 L L 1 1 L 60 70 80 90 100 110 120 130 DAYS POST TREATMENT Figure A-18 61 Figure A—l9. Per cent of total applied copper present in various components of NW pond from time of dosage until end of experiment. PER CENT OF TOTAL COPPER APPLIED 100 80 60 40 20 20 62 r—O NW Water g"... NW Sediment o—o NW Plant Tissue 0"... L J l - 30 40 50 60 DAYS AFTER TREATMENT Figure A—l9 SUMMARY Copper dispersal was characterized by an immediate active uptake by plant tissue and by a rapid precipitation, probably of malachite. Sediment accumulated copper rapidly at first, probably from precipitates, adsorption of ionic or complexed forms, and decaying plant tissue. As a gradual decline to background c0pper levels in water occurred, organic compounds, perhaps humic acid-c0pper complexes, may have precipitated or been adsorbed by the sediment. Adsorp- tion would explain the gradual increase in copper levels in sediment. Fish and snails, while relatively unimportant in total copper dispersal due to small total mass, accumulated copper. In fish, levels were low, and copper was eliminated from fish rapidly. 63 PART II EFFECTS OF COPPER ON FISH AND FISH-FOOD ORGANISMS 64 INTRODUCTION Toxicity of copper to aquatic organisms is well estab- lished. Extensive work has been done in laboratory studies to determine not only acute toxicity limits but also subtle effects of copper on such processes as growth and reproduc- tion. Volume 3 of the Water Quality Criteria Data Books, Effects of Chemicals on Aquatic Life (1971), summarizes past copper toxicology work. Field studies have been less frequent. Several workers (Riley, 1939 and Hutchinson, 1957) have speculated about possible toxic effects of natural c0pper levels. Others (Smith, 1939 and Eipper, 1959) have added copper, usually as c0pper sulfate, to aquatic systems and noted effects on organ- isms. Few efforts have been made to relate laboratory bio- assays to subsequent field work. In the present study, tolerance limits were established for common pond organisms in bioassays. _Then pond systems containing these or related organisms were dosed with copper sulfate, and effects on fish and their food organisms were studied. 65 MATERIALS AND METHODS Laboratory Studies A series of static bioassays were performed to determine c0pper tolerance of organisms found in outdoor ponds. Among organisms tested were a cladoceran, Daphnia pulex; a copepod, CycloEs sp.; two dipteran larva, Chaoborus sp. and Chironomus tentans; and green sunfish, Lepomis cyanellus. COpper toxicity is known to fluctuate widely with alka- linity changes. To maintain similar alkalinity between indoor and outdoor studies, all tests were conducted in water pumped from the outdoor ponds. Conditions listed by Standard Methods for the Examination of Water and Wastewater (1965) were fol- lowed for the bioassays. Replicate tests were conducted. Invertebrate bioassays lasted 96 hours, while sunfish were tested for 30 days. Water in fish tests was changed every other day to prevent possible effects of fish wastes on copper toxicity. A growth study was conducted on green sunfish. Total weight of fish in each test container was determined at the beginning of the test. Ten per cent of this weight in live cladocera was fed the fish every other day. At the end of the test final weights of surviving fish were determined to assess effect of copper on green sunfish growth. 66 67 Pond Studies Zooplankton was sampled at four stations established in each pond. In SW, NW, and NE ponds, with their extensive macrophyte growth, two stations were established in weedy areas and two in clear areas to account for differences in population densities. In SE pond, since few macrophytes existed in the beginning of the test, one station was estab- lished near the outside, one near the center, and two in the middle of the pond. Vertical tows with a Wisconsin plankton net collected the samples. Organisms were rinsed into a sample bottle and preserved with formalin. Enumeration of zooplankton was done in two ways. Total sample counts were performed on large forms, like cladocera, ostracods, and copepods, while smaller forms, like rotifers, were enumerated using a Sedgwick-Rafter counting cell. Complete counts of larger forms were necessitated by their low density. Benthos was sampled with an Ekman dredge. Three samples were taken each sampling date from the inside, middle, and outside of each pond. After collection, mud was rinsed from each sample; sugar flotation (Anderson, 1959) was used to sort the Specimens which were then preserved in formalin. Freshwater Invertebrates of the United States (Pennak, 1953) was used to aid identification of zooplankton and benthos. Confirmation of certain forms was given by Dr. Donald McGregor (ostracods), Dr. Harry Yeatman (cladocera and copepods), and Dr. Clarence Goodnight (aquatic oligochaetes). Table B¢l lists organisms common in the ponds. Fish were captured in baited traps, blotted dry, and weighed. Fish were released after each weighing. Several specimens, however, were sacrificed for stomach and copper analyses. After c0pper sulfate dosage all dead fish were collected and recorded. At the end of the tests the ponds were drained, and surviving fish were weighed. Table B-1. Organisms common in outdoor study ponds. CLADOCERA: Scapholeberis kingi Diaphanosoma leuchtenbergianum Egydorus sphaericus Simocephalus vetulus Pleuroxus denticulatus Ceriodaphnia reticulata Daphnia pulex (rare) COPEPODA: Diaptomus pallidus Mesocyclops edax Macrocyclops albidus Eucyclops agifls Cyclops vernalis Cyclops varicans rubellus OSTRACODA: 9%pridopsis vidua P ysocypria pustulosa (rare) ROTIFERA: Keratella sp. Platyias sp. Lepadella sp. Monostyla sp. Lecane sp. Philodina sp. OLIGOCHAETA: Limnodrilus hoffmeisteri Limnodrilus profundicola Potomothrix vejdovsKyi CHIRONOMIDAE: Chironomus tentans RESULTS AND DISCUSSION Laboratory Study Bioassay The cladoceran, Daphnia pulex, was almost 1000 times more susceptible to copper than the copepod, Cyclops sp. (Table B-2). Chaoborus sp. and Chironomus tentans were resistant to high levels of copper, while green sunfish displayed a reSponse intermediate between the extreme sensitivity of cladocera and the relative insensitivity of other organisms tested. Other workers have confirmed the toxicity of copper to cladocera in bioassays. Fisher (1956) noted that 2 ppm copper killed Daphnia pulex within 72 hours, while Cyclops sp. was not affected by 5 ppm c0pper. Rose (1954) noted that Cyclops sp. and Diaptomus sp. survived 5 ppm copper sulfate indefinite- ly, while Daphnia sp. was dead after 20 hours in 0.5 ppm copper sulfate. Riley (1939) noted that Chaoborus survived 200 ppm copper for 124 hours in a bioassay. He speculated that an inpene- trable cuticle was responsible for their tolerance. Fish vary in their responses to copper. O'Hara (1970) noted that concentrations of copper ranging from 0.01 to 10 ppm have been reported as 96-hr TLm values for fish (a 96-hr 69 70 Table B-2. Bioassay results. Organism Length of TLm (ppm copper) Test Replicate 1 Replicate 2 Cyclops sp. 96 hour 27.25 24.50 Chaoborus sp. 96 hour >225.0 >225.0 Daphnia pulex 96 hour 0.028 0.028 Chironomus tentans 96 hour 35.25 26.0 Lepomis gyanellus 30 day 1.68 2.0 TLm value is the toxicant level at which half the test organ- isms survive for 96 hours). TLm values of 1.68 and 2.0 ppm copper found for green sunfish in the present study are similar to past values for centrarchids. Cope (1966) gave a 96-hr TLm value of 2.8 ppm copper for bluegills. Pickering and Henderson (1965) found a 96-hr TLm of 0.66 ppm copper for bluegills tested in soft water. Cairns and Scheier (1968) reported a 1.25 ppm copper 96-hr TLm value for bluegills. Since chemical parameters often differ in bioassays and species differences may occur between such organisms as blue- gills and green sunfish, limited inferences can be drawn from past bioassay results. Apparently, low levels of copper used in the present pond study should be toxic only to cladocera among the organisms tested. 71 Growth Study Green sunfish exhibited differences in feeding habits. Fish in the control and two lowest c0pper concentrations fed actively on cladocera, while those at 1 and 2 ppm copper ingested few cladocera and often regurgitated any specimens eaten. Effects of the decreased feeding activity failed to appear in growth differences. Fish in all copper concentra- tions and the control lost weight (Table B-3); none of the growth differences between concentrations in either replicate were significant at the 5% level when tested with Tukey's t-test. Table B-3. Weight changes in green sunfish during growth study. Copper treatment Number of fish (ppm copper) surviving test Mean % wt change i} 1 S.E. (out of 10) Rep. A 0.00 10 -16.83 1.615 0.125 10 -22.47 1.270 0.25 9 -20.31 1.930 0.50 6 -22.68 2.610 1.00 7 -22.82 2.710 2.00 5 -24.20 2.034 Rep. B 0000 10 ‘20085 10090 0.125 10 -20.98 1.858 0.25 10 -22.34 0.948 0.50 10 -25.32 1.466 1.00 7 -25.00 1.867 2.00 2 -19.40 0.300 1‘. 72 Mount (1968) and McKim and Benoit (1971) noted a reduc- tion in fathead minnow and brook trout growth respectively at low copper concentrations in laboratory bioassays. Comparison of fish growth studies is difficult because of different ex- perimental techniques. Pond Studies Zooplankton Zooplankton samples from weeded and clear areas in NW, SW, and NE ponds are graphed separately (Figures B-l, B-3, and B-5). Organisms from all four sampling stations are . lumped in SE pond (Figure B-7). SW Pond COpper sulfate treatment caused a precipitous decline in cladocera in SW pond (Figure B-l), while levels remained constant in the control pond (NW). Rotifers also disappeared from SW pond samples. Ostracods and copepods showed no appar- ent effects from the copper; the decrease noted in copepod numbers near dosage time occurred in a pre-treatment sample. Cladocera and rotifer populations in SW pond recovered rapidly from copper sulfate dosage; 4 weeks after treatment, both populations began to increase. A possible mechanism for rapid repopulation by cladocera was the constant release of individuals from ephippia. Those released when the copper concentration declined to a non-lethal level repopulated the pond. 73 Figure B-l. Changes in SW pond zooplankton numbers over time. Each point represents the average of two samples from either weeded (solid line) or clear (broken line) areas of the pond. Shaded area denotes addition of 3 ppm copper sulfate. ORGANISMS/LITER 100 60 74 Rotifers Ostracods TIME Figure B-l 75 Dissolved copper concentration during cladoceran reap- pearance was above toxicity limits delineated in laboratory bioassays (see Figure A-l). Possibly dissolved copper was organically bound and rendered less toxic. Stiff (1971) noted that humic substances reduced acute copper toxicity to fish by complexation. A similar mechanism may have operated * in SW pond. All four groups of zooplankters studied experienced population increases after dosage. Peak numbers were noted first for rotifers, next for cladocera, then copepods, and finally ostracods. This pattern was not observed in other ponds. Decay of plant tissue may have given rise to bacterial and protozoan populations which acted as food sources for rotifers; increases in all three groups may have then supported population increases among cladocera and c0pepods. Decline after treatment occurred among all four major species of cladocera (Figure B-2). Each species recovered and underwent a late-season population rise. This coincidence of peaks was not noted in the control pond and may have been a delayed response to interruption by copper of normal popula- tion patterns earlier in the season. NW Pond Both cladocera and rotifers were immediately affected by the c0pper with populations declining to near zero levels (Figure B-3). Rotifer and cladocera populations in SW pond were moderately high at this time. 76 Changes in numbers of SW pond cladoceran species over time. Each point represents the average of two samples from either weeded (solid line) or clear (broken line) areas of the pond. Shaded area denotes addition of 3 ppm copper sulfate. ORGANISMS/LITER 30 _ - 9. 10- 3 3 3.0 ~ 10 ”‘ as". .....O 3.. . f‘ 30 f‘ : .1 :‘2: 1° - 5 3’? o,,....‘ um i 60 20 5-31 6-20 7-10 77 Pleuroxus denticulatus Chydorus sphaericus fl... ooolfl'.‘ ooooloooooooo . P m m. 0...... Simocephalus vetulus 0“. 0". ‘ Ccriodaphnia reticulata 10-18 Figure B-2 78 Figure B-3. Changes in NW pond zooplankton numbers over time. Each point represents the average of two samples from either weeded (solid line) or clear (broken line) areas of the pond. Shaded area denotes addition of 3 ppm copper sulfate. 1500 500 30— 10 -’ '79 Ostra00d3 00....-.000 ...: CopePOd‘ “ :: Ill : '0 a g g H . . d 5 "’ 3 so —- E 3 L5. : -. a o z 20 5 . ..-.. 5 a °‘° o-W" 0‘ 0. on... . ..... 125 -- Clad“era 7s — ' P 25 . . .0. 0.0 O. ’0 . ..‘... . ...... ‘ 00.. .0. O... .000 O... o .. "a“... 1 n 1 ° ”u- 1 -m* 7-20 8-9 8'29 9-18 6-10 5-30 Figure 3’3 :OCW 10-28 80 Possible indirect effects of copper were indicated in NW pond when copepods disappeared 4 weeks after treatment; a drastic reduction in oxygen and release of H S following 2 decomposition of Chara may have been responsible. Figure B-4 shows individual numbers of cladoceran species in NW pond. The decline noted in NW pond was due largely to decreases in Chydorus Sphaericus numbers. NE Pond No definite effects of copper appeared in NE pond zoo- plankton numbers (Figure B-5). A decrease from 30 to 15 cladocera per liter of water occurred after treatment. The two major species present during dosage were Chydorus sphaericus and Pleuroxus denticulatus. Numbers of each declined slightly but increased to a smaller peak after 3 weeks (Figure B-6). The suppression may or may not have been due to copper. SE Pond Sharp declines in both rotifer and cladocera populations were noted in SE pond after an initial 1 ppm copper sulfate dosage, although neither population was reduced to zero levels (Figure B-7). Cladocera recovered to pre-treatment population numbers before again declining to near zero after a second treatment of 1 ppm. Populations in the control pond (NE) underwent no such sharp fluctuations during this period. Copepods did not appear to be affected by the copper applica- tions. Figure B-4. 81 Changes in numbers of NW pond cladoceran species over time. Each point represents the average of two samples from either weeded (solid line) or clear (broken line) areas of the pond. Shaded area denotes addition of 3 ppm copper sulfate. ORGANISMS/LITER 10_. 82 Ceriodaphnia reticulata Scapholeberis kingi Simocgphalus vetulus 30 20— Chydorus sphaericus .IOI.‘......|IOIOI.:L ’00.....‘.o Pleuroxus dentIculatus 7-20 8-9 8-29 9-18 10-8 10-28 Figure B-4 Figure B-S. 83 Changes in NE pond zooplankton numbers over time. Each point represents the average of two samples from either weeded (solid line) or clear (broken line) areas of the pond. Shaded area denotes addition of 1 ppm copper sulfate. 600 200 60 20 0| ‘0 I 50 84 ORGARISNS/Lxran I N O 3o __ 10 __ Ostracods IO.0.00.00000000000.‘........ .:.:.. COpepoda A --;_---_‘ Cladocera ififi 9-6 9'26 10-16 Figure B-6. 85 Changes in numbers of NE pond cladoceran species over time. Each point represents the average of two samples from either weeded (solid line) or clear (broken line) areas of the pond. Shaded area denotes addition of 1 ppm copper sulfate. 86 Chydorus sphaericus SO nnnn I 30 ..... ORGANISMS/LITER I.- .. Il.‘ .OOOOOOIOOOIOOOOOIOIICCC '.'.'. o .......... ...... . ooooo ...... ...... g Pleuroxus v ent culatus 6-15 7-5 8-17 9-6 9-26 10-16 11-5 11-25 Figure B-6 87 Figure B-7. Changes in SE pond zooplankton numbers over time. Each point represents the average of four samples from the pond. Shaded areas denote additions of l, l, 3, and 5 ppm copper sulfate. 88 1000 _— 60k 20f o 5 140%— 100_— 600 r- 200 - 1505- ”fiyfiigaflzaio 7-8 8-17 9-6 9-26 10-16 11-5 11-25 6-10 TIME Figure B-7 89 Differences in reaction to copper existed among various species of cladocera in SE pond (Figure B-8). The two major species present were Diaphanosoma leuchtenbergianum and Scapholeberis kingi. Diaphanosoma disappeared rapidly after the initial dosage, while Scapholeberis appeared unaffected by the first dosage but decreased after the second applica- tion. The decline exhibited by Scapholeberis was more gradual, occurring over a 2-week period, and may not have been attribut- able to copper. Field data concerning the effects of copper on zooplankton vary. Smith (1939) noted that in four Nova Scotian lakes, cladocera were susceptible to 3 ppm c0pper sulfate, while cer- tain copepods were present in reduced numbers for some days following treatment. In his study about 1 year elapsed before planktonic crustacea became numerous in net hauls. Water in these lakes was acidic and maintained high copper levels for a long period. Rose (1954) treated the upper 3 feet of Storm Lake, Iowa, with 1 ppm c0pper sulfate several times throughout the summer; after the second treatment, all Daphnia were dead, but Cyclops were found in concentrations of 45 per liter of water. Crance (1963) found that in ponds treated with about 0.4 ppm copper sulfate, c0pepods were the dominant zooplankters but that some cladocera were present. Little data exists con- cerning effects of copper on ostracods or rotifers. 90 Figure B-8. Changes in numbers of SE pond cladoceran species over time. Each point represents thepaverage of four samples from the pond. Shaded areas denote additions of 1, l, 3, and 5 ppm copper sulfate. 91 _ [WWI . aw." MP... .959? titan-Ono. trio-Dona... nnnnnnnn a... 3.4.»... u. .Awn... ..... ..... ....... 2 I 100 *"' 60 20 _ o 4 1 anaHQ\umez¢u¢o 100 _- 60 20 TIME Figure B-8 92 Results of past field work and the present laboratory bioassays seemed only partially confirmed in the present field study. SW and NW ponds showed a marked decline in cladoceran and rotifer populations, but a fairly rapid repopulation occurred in SW pond. Results in NE and SE ponds indicated that some species differences may occur at low copper levels. Both ponds were originally subjected to 1 ppm copper sulfate; in SE pond, Diaphanosoma leuchtenbergianum was apparently quite suscept- ible to the dosage, while Scapholeberis kingi was less - affected. In NE pond, Chydorus sphaericus and Pleuroxus denticulatus showed only slight numerical decreases. Other natural factors, including age distribution and condition of the specimens, may have been involved in these population fluc- tuations. At copper sulfate levels used for algal control (0.25 to 1.0 ppm), it is difficult to predict effects on cladocera, as species differences may be involved. The higher doses recom- mended for macrophyte control appeared toxic to all cladoceran species, as illustrated in SW pond. Suppression was short- lived, however, as cladocera reappeared within 4 weeks. Rotifers were susceptible to both dosage levels used (1.0 and 3.0 ppm), while c0pepods and ostracods were not directly affected at any copper concentration. Indirect toxicity to all groups may occur when large masses of plant tissues decom- pose, as in NW pond. 93 Benthos Aquatic oligochaetes and chironomid larva were dominant members of the benthos. Snails, mayflies, odonates, and other dipteran larva were present in smaller numbers (Tables B-4 through B-7). No changes directly attributable to copper were noted among any organisms in SW, SE, or NE ponds. Population de- creases did occur in NW pond, however (Table B-4). Oxygen stress and high levels of H S following Chara decay may have 2 been responsible. Chironomid larva numbers declined from about 3000/m2 to approximately ZOO/m2, while aquatic oligo- chaete numbers changed little. Snails, mayflies, and other less numerous members of the benthos were not found during the final four sampling periods in NW pond. Only haliplid larva occurred in these samples, all of which were taken after decomposition of Chara began. Previous work on effects of copper on benthic organisms has been inconclusive. Walker (1961) noted that copper sul- fate significantly reduced bottom fauna during a l—year period following treatment. Smith (1939) found in Nova Scotian lakes that treatment with 3 ppm c0pper sulfate decimated benthic organisms; insect fauna reestablished themselves fairly rapidly, while molluscs did not. Sohacki (1965) noted significant reductions in all benthic species, including chironomids, may- flies, odonates, and molluscs, when he dosed a pond with 94 .mmsm pflsocouflnom ampfiasaamnflqo um>HmH pwawnmoupmmm m>umH mappmom um>uma vasomomoumumuo um>uma .mm mononomnuo um>uma pflamflammm “.mm mnszan¢ .mm MHHmHmEmnmmm «.mm mwcmmmxmmm “.mm mflsmmos «.mm omNnmm “Hum magma mwmm “Hum magma momH mvH Hum mom OHIHH mva mud mw haloa mama and mauoa mva mnH mam OHIOH llllllllllllllflllldlllfl llllllllllllllllllllllllllllllllllllllllllllllllllllllllllll .... lllll mvH.Mmm.ovH.m¢H :vH mam mvm mmmm vm m «me va OMH mom mum 0mm.MmN.¢vH mm com mom mmlm mmvu<>m Hmm mv hmH malm mmv.wmm.mvfl.m mucmmwumwu Hones: zoom .mEHu Hm>o coon 32 ca E\mEmwcmmHo Dazuswn mo muwnasc sfl momsmno .vum manna N am>uma paomauzo 95 m am>HMa vasomOQOpmumo O um>HMa .mmsm pacomomoumnmo m .mm msuonomno U “mcaasaambaqo um>uma nauosammm am>uma paamaammm “.mm muscnma < .mm macmmus “.mm mmNnmm “aim magma momN “aim manna momH Gama mmm mnma oalaa awn mom mmaa mmuoa ova.amv.0va.o paom 3m ca Hones: comm N .mmamEmm mounp mo mmmum>m mucmmwumou E\mEmacmmHo oazusmn mo madman: ca mmmcmnu .mlm manna .mm muosnmam am>uma paomaumom .dm mmxsm. uTm magma 6mm. Nanm magma omm. mmh maIaa mvm oana va maIm MIm mmm va mNIm mva maIm dva mm amp aMIn vmv nmIm Nmmpmmnoomaao msomcmaawomaz mwaammmz mmaamnm oaumow< flmmaeoconano mama .wummaSm Hmmmoo Ema m mam .m .a .a mo mcoapammm muocmm mmsaa III was .mmamfimm mounu mo mmmnw>m mucommummu Hones: comm .mEau Hm>o moon mm ca NE\mEmacmmHo oaaucmn mo mHmQEs: ca mmmcmno .mlm magma 97 «.mm mass m o m mam .mm macmmus a.mm mmmnmm aaIm magma mmm~ «aIm magma momfl .m>nma pacomomoumumum “.mm macannuoosma am>uma maamaammo am>uma mampmo mva maa mama mm maIaa Mva.0va.mnm mm moma mIaa Unmudva :va oaoa mmva mam maIm ava.0mm.4va 0mm mama maa mIm mam mmoa vva mmIm mmm mom aaaa maIm mum movm mv amIn mm mmma mmIm «mm am aom hmIm Nmmummzoomaao msomcmaamomaz mmaammmz mmaamcm oaumswm Hmmaeoconanu mumo .mummamm uwmmoo 5mm a mo coauapmm mmuocmp mcaa III map 30amm .mmamamm mmunu mo mmmuw>m mucmmmumwu H0985: 30mm .maau um>o moon mz ca E\mEmacmmHo oasucmn mo mnwnfis: ca momsmnu .hIm magma m 98 2 ppm copper sulfate. Mackenthun and Cooley (1952), however, performed bioassays using copper mixed with sediment and found that such benthic organisms as Tendipes plumosus and Pisidium idahoense could tolerate up to 9,000 ppm c0pper based on a mg copper/kg dry sediment basis. This method of expressing copper toxicity is questionable, however, as much of the copper would be bound to the sediment and unavailable to the organisms. Laboratory bioassays have shown that certain benthic organisms are intolerant of copper. Warnick and Bell (1969) showed a 48-hr TLm of 0.32 ppm copper for the mayfly, Ephemerella subvaria. Arthur and Leonard (1970) found an aver- age 96-hr TLm of 0.039 ppm copper for Physa integra and 0.02 ppm copper for Gammarus pseudolimnaeus. Although insufficient numbers of mayflies or snails existed in the present study to permit conclusions, these organisms may have been eliminated by dosages of 3 ppm copper sulfate. That their loss affected fish growth is doubtful, however. No evidence of direct c0pper toxicity to either chironomid larva or aquatic oligochaetes existed in any pond. A more serious potential danger to benthic organisms, as illustrated in NW pond, is massive disintegration of plant tissue with its concomitant oxygen stress and release of de- composition products. Synergistic effects of low oxygen and H S may have caused the decline in chironomid populations in 2 NW pond. 99 Fish Direct Toxicity No green sunfish deaths directly attributable to copper were noted in this study. As dosage rates were below labora— tory TLm values, the absence of mortality was not surprising. Evidence of direct copper toxicity at low concentrations in ponds is scarce. Eipper (1959) noted that small golden shiners, Notemigonus crysoleucas, were killed in a pond by 1 ppm copper sulfate. Smith (1939) found that 3 ppm copper sulfate killed both white perch, Morone americana, and yellow perch, Perca flavescens. Indirect Toxicity Indirect toxicity did occur in NW pond. Only four fish survived until the end of the experiment in this pond. Although oxygen stress occurred in the NE, SW, and NW ponds, it caused no mortality in either NE or SW systems. In NW pond, however, after large masses of EEEEE were killed by copper, resulting decomposition caused almost complete fish mortality. Oxygen levels were below 0.5 ppm for 10 days; c0pper concentrations rose to about 0.5 ppm; and the odor of H25 was present. Although the fish were characteristically gulping air during the first days of stress, they soon ceased this activity. COpper, H S, or low oxygen probably acted 2 singly or additively to cause mortality. 100 Other herbicides have caused fish deaths indirectly. Surber and Everhart (1950) treated a series of ponds with 1.2 ppm nigrosine (a black aniline dye) and noted that some kill of smallmouth bass occurred because of suffocation caused by decaying Chara. Fish Growth NW and SW Ponds The effects of c0pper application on fish growth in NW and SW ponds are shown in Figure B-9. Each point represents the mean of per cent weight increases calculated for all fish captured at a sampling period. Fish were released, and a new sample was captured for each weighing. Sample sizes ranged from 7 to 42 in SW pond and 5 to 33 in NW pond. Using logs of the data, 95% confidence limits were placed around each mean. Scheffe's test was used to compare means. Growth in the treated pond (SW) was greater than in the control pond (NW) after the third week. Three areas of the fish growth curves will be discussed. First, no significant differences in growth occurred between the two ponds in the initial 3 weeks following treatment. Stomach analyses showed that while cladocera disappeared from the diet in SW pond after copper sulfate treatment, other prey organisms were fed upon (Table D-3, Appendix). Large numbers of copepods and chironomids along with lesser numbers of other 101 - Figure B-9. Mean per cent weight increases for green sunfish in NW (solid line) and SW (broken line) ponds over time. First shaded area denotes addition of 3 ppm copper sulfate to SW pond, while second shaded area denotes addition of 3 ppm copper sul- fate to NW pond. MEAN PER CENT NEIGET INCREASE 1100 , 900 b 7001- SOOr' 300 l 100- W 95% Confidence Limits on NW Means .+-o 95% Confidence Limits on SW Means *- ..... a ..... ----- ..... a ..... ..... ..... ..... ..... ----- ..... IOOI av.- ..... ..... ..... s ..... .... ..... ..... ..... IIIII ----- ..- ..... ..... ..... ..... ----- ..... ..... ..... .wg. ooooo one. ..... ..... ..... ----- ..... ooooo '''''''''' 102 Denotes Significantly T Different Means (5% Level) 11" U -- n ...... --.. ..... ... ..... ...- ooooo .... ..... ...- ..... .c u ..... ......... ..... noun ..... ..... uuuuuu IIII ......... ..... Figure B—9 I 11-21 103 prey were found in fish stomachs. Apparently neither loss of cladocera nor possible copper stress adversely affected growth rates of fish in SW pond immediately after treatment. Three weeks after treatment with c0pper, fish growth rates increased in SW pond more rapidly than in NW pond. The three sampling periods following the initial 3—week phase showed significantly higher mean growth rates in SW than in NW pond. The substantial increase may have involved two factors: (1) since spawning occurred in NW pond during this period while none took place in SW pond, energy used by fish for reproduction in NW pond may have suppressed their growth rates; and (2) loss of macrophyte cover may have caused prey to become more susceptible to fish in SW pond. Tompkins and Bridges (1958) found above-average fish productivity in Lake Quannopowitt, Massachusetts, despite c0pper treatments over long periods of time. Gilderhus (1966) found that bluegill growth rates were higher in ponds treated with low levels of sodium arsenite than in control ponds; he noted that succession of plankton blooms was more rapid in lightly-treated ponds than in control situations. A rapid decrease in growth was noted after copper sulfate dosage in NW pond. Oxygen stress and adverse reactions to H28 and copper were probably responsible for the decline. NE and SE Ponds Small sample sizes of fish from NE pond and multiple copper sulfate dosages in SE pond made analysis of growth 104 difficult. As all but the last two samples in NE pond served as controls, effects of c0pper on fish growth in SE pond ap— peared insignificant; growth rates of fish in both ponds remained fairly similar throughout the test (Figure B-lO). Fish Reproduction In August, fish fry were seen in NW pond, but none ap- peared in SW pond. The 3 ppm copper sulfate dosage in SW pond may have interferred with spawning activities. Lack of fry in SW pond may not have resulted from physiological effects of copper but from destruction of plant tissue. The oxygen depletion caused by decaying tissue may have been lethal to any eggs or fry present; physical disruption of the nests by dying plants may also have occurred. Figure B-lO. 105 Mean per cent weight increases for green sun- fish in SE (solid line) and NE (broken line) ponds over time. First three shaded areas denote additions of l, l, and 3 ppm copper sulfate to SE pond, while the last shaded area denotes addition of 1 ppm copper culfate to NE pond. MEAN PER CENT WEIGHT INCREASE 900 .— 700 - 500 )— 300 100 106 O-‘III 95‘ Confidence Limits on SE Means H4 95% Confidence Limits on NB Means -.-:;:§:§:l:;:;: . . 5.; ... . I C s': ‘u:-:c‘ 1:3:1: . I C . non- ......... "Pi-1'5, . .. ' '.3'3:1:-. . . .'.'- . I. Exec-r ' .- .-:-:-: a... ..... ...- O I O I . g 1.;.;.;.;.;.'.;.;.-.;.;.;.:.; -: --:-:-:-:-:-$'-' 3 ' - ' Figure B-lO PI. SUMMARY Effects of copper on fish and their food organisms de- pended on dosage level. At 3 ppm c0pper sulfate, cladocera and rotifers among zooplankters were temporarily eliminated but population numbers increased within 4 weeks. At 1 ppm copper sulfate, less spectacular decreases were noted and species differences existed among cladocera in their responses to c0pper. Copepods and ostracods were not directly affected at any copper sulfate level. Effects on benthic organisms were difficult to detect. Decreases occurred in chironomid larva and less numerous popu- lations in NW pond after dosage and were probably due to in- direct effects, such as low oxygen and release of copper and H28 from dying Ehggg. Fish deaths were limited to apparent indirect toxicity in NW pond, while green sunfish in the treated SW pond had higher relative growth rates than fish in the control pond. A decrease in reproductive products in SW pond may have in- creased energy available for growth. 107 PART III EFFECTIVENESS OF COPPER SULFATE AS AN AQUATIC HERBICIDE 108 INTRODUCTION The effectiveness of copper sulfate as an aquatic herbi- cide has been widely documented. Mulligan (1967) stated that c0pper sulfate was the most widely used aquatic herbicide. Phelps Dodge Refining Corporation, manufacturer of copper sul- fate, recommends 0.25 ppm copper sulfate for planktonic algae control, while 0.5 ppm is recommended for filamentous algae control. Sutton and Blackburn (1971) stated that a concentration of 1.0 ppm copper or less controlled most species of algae. They noted, however, that some species, such as Pithophora sp., were resistant to copper sulfate and required repeated doses for control. Fitzgerald (1966) found a wide variation in susceptibility to copper sulfate among algal groups; certain species, like Microcystis aeruginosa and Anabaena circinalis, were killed by 0.1 ppm copper sulfate, while Dictyosphaerium pulchellum survived 8.0 ppm for 72 hours. Smith (1939) found that 3.0 ppm copper sulfate in Nova Scotian lakes completely destroyed algal populations; often forms were eliminated for up to 3 years.’ The unusual duration of control in this study was probably due to the maintenance of high copper concentrations in the acidic water. 109 110 Copper sulfate is increasingly used for aquatic macrophyte control. Phelps Dodge recommends doses of from 5.0 ppm to 10.0 ppm c0pper sulfate in water with alkalinity of less than 50 ppm, and 40.0 ppm copper sulfate in moderate to hard waters for macrOphyte control. Sutton and Blackburn (1971) noted that concentrations of copper sulfate at 112 to 560 kg/ha were necessary to control submerged species, while emergent species seemed resistant to copper treatment. Ware (1966) found that copper sulfate at 50 lbs/surface acre gave 75% control of Elodea densa; at 100 lbs/surface acre, control was complete. Huckins (1960) applied copper sulfate at 500 lbs/acre and controlled Potamogeton crispus and Elodea canadensis in New Jersey lakes. Chancellor e3 El- (1958) found that a constant concentration of about 0.25 ppm copper in a stream eliminated Elodea canadensis, Potamogeton spp. and Lemna minor after 8 weeks. Smith (1939) found that Nymphaea, Juncus, Pontederia, Scirpus, Eriocaulon, and Potamogeton were not affected by copper sulfate concentrations of 3 ppm. In Part III the effectiveness of copper sulfate treat- ments in algal and macrophyte control is investigated. Both negative and positive aspects of copper sulfate control are documented. MATERIALS AND METHODS Phytoplankton samples were collected at the four stations used for zooplankton sampling in each pond (see methods section of Part II). A water column collected in a core sampler was subsampled, fixed with formalin, and passed through a membrane filter. The filter was cleared with immersion oil, and phyto— plankters were enumerated (McNabb, 1960). Direct observations were made on the condition of filamentous algae and macro- phytes throughout the study. Water chemistry parameters are shown in Figures A—l through A—4. The schedule and rates of c0pper sulfate dosage are shown in Table A-1. 111 RESULTS AND DISCUSSION The dominant species in NW and SW ponds was Ceratium hirundinella (Tables C-1 and C-2), which underwent a 90% reduc- tion in NW pond but increased in SW pond after dosage with copper sulfate. In NE pond (Table C-3) the dominant species, Trachelomonas sp., suffered no effects at 1 ppm copper sulfate. In SE pond (Table C-4), both Ceratium and Trachelomonas were present, but little effect on either population was apparent. Consistent control of dominant species in the ponds, Ceratium and Trachelomonas, was not attained. Control of filamentous algae in all ponds was complete. In SW and NW ponds, Oedogonium sp. was controlled at 3 ppm copper sulfate. In NE pond, 1 ppm copper sulfate gave good control of Spirogyra sp. and Oedogonium. Chara sp. was com- pletely eliminated in NW pond, but damage was slight in NE pond at 1 ppm c0pper sulfate. A blue-green periphyton growth on the walls of SE pond was not visably affected by l, 3, or 5 ppm copper sulfate. Otto (1970) noted that blue-green algae mats were more re- sistant to copper sulfate treatments than were filamentous green algae. He hypothesized that the gelatinous sheaths of algal filaments may have reduced activity of copper sulfate. 112 113 .mmmnm pmpmmz Scum mmamEmmN .mmmnm Hmmao Bonn mmamEmm_ wma mmv 5mm vvm mnm ava hr mmloa am pm mom mmm momh mwmm om mva mow mmm haloa maoa mhoa omam omma II ov mam ow oaloa om II om mm mmmv homa omIm we we mo II wmmvv ovvma mem wv mm II we hvomomooomam malm mmha haaN vlm momhm II mmlm .faom hmoma hmmaw malm mm II hmmoa mmmma aaIm om II ov vmmma II vlm ma ova 0v II ammmv mmlh mm II mmmma mmmmm amlh om am mm mmvmn hmmvm main II ma hmaam mmvam main II mmomm hmahv malh mm mm omhmw ahmmm aalh II mm amh omaa vbho Nvmv mmlm am II II No mma mvh NNIm mm mm am II II voa mmmm malw II ma mm mm II vb mam Nmm calm z o z o z o z o z o 3 o N: .o maummo msomnm mamamsm EDaHmEmoo EdanmumOaU mommnmcm Edaumnmw mumo IomOaU .mummasm Hmmmoo Ema m mo coauampm mmuocmp mcaa III may 30amm .mmmum sz pmpmmz Scum cam ADV Hmmao Scum pmmmum>m mum3 mumo 50mm :0 mmamfimm 039 .mfiau Hm>o umuaa\souxsmamoumam pcom 32 mo mumnfid: ca mmmcmnv .aIU magma 114 .wmmnm mmpmm3 scum mmamEmmN .mmmum Hmmao Eoum mmamammH mm mm mamv amhm mhma 5mm aana mmmmb mm mhva mow ana mm anm own amma mbmm mmlm II mom maIm mo vow va mwm mmm mmm aaIm mm mmm mmm ma va om om om mma mm mth ma mm mm mm mm mmm omb amlh ha mm new voam malh ma ova ma mbmm vmma maln mm om mwmm omm vth mm hva ma mmma mmm maIn ma mna ha ema mob «mm aalb mm awn II mmIm mam moa am am mvm aoma ammm mmIm mm om mm mm om ea comm amm cam maIm am mm ma calm 3 U 3 U 3 o 3 U 3 U 3 0 N3 mu maumNo msomsm mcmammm Emanmamou Edaumuman mammnmc< Esaumumu mumo Iomoaw .mummasm Hmmmoo Ema m mo coauampm mmpocmp mcaa III ms» 30amm .mmmum A3V mmpmm3 Eoum mam ADV Hmmao Sony pmmmum>m mum3 mump nomm so mmamfimm 039 .mEau Hm>o Hmuaa\c0uxcmamou>£m pcom 3m mo mnmneas ca mmmcmnu .mIU magma 115 mm m oa wmmmnmm malaa em ma om mnavmh oaIaa om mm om oa mvoomm aIaa m hmmmm mmloa mm mammm mana am m ommoow maIm om mamma mIm mom II ommma hmIm on mom II mmIm vwa mma II balm ow mamm aMIb mm am monm mIn up II mvmmmmm mmIm mmmmom ammaomm malm mmmmmmm mmmmmm Emaamfimoo EaaamamOau mmaoeoamaomaa mammamaa mama .mammasm Hmaaoo Ema a mo coaaappm mmaoamp maaa III maa 30amm .mamp nomm so Ummmum>m mHm3 mmaaEmm asom .mEaa am>o Hmaaa\soaxamaaoa>aa pcoa mz mo mumafiac Ga mmmsmao .MIU magma 116 am II om maIaa m II II maIaa mmaa ha Mlaa ov vbwmm ow amIoa mm mmmoom omv oana mmhmmm mm mIoa om mmmvh om omlm vm mmommm ham mmIm Noawmm mhma malm mm mmmmva mm maIm mm mmm mm mmomoma amma mIm mma ha mmmmwm aha hmlw mmmmmn mam mmlm ma mwmmoa momm omlm movmam 5v malw omovam hmm baIm bawhwm II valm om mmmonm oovoa amIb am mammoa «vmma mlh om II nma mwmommma mmIm mm ma II ham aovmmmm oalm mamamsm mammaa Edaamamou mm:OanmaumaB Enaamamu mammamcm mama Ema m mam .m .a .a mo meaaappm maoamp mmcaa III maB mmaaEmm uaom .mEaa Hm>o Hmaaa\aoaxcmaaoa>aa paoa am no mumafiac ca mmmamao .mammaam amaaoo .mamp 30mm so pmmmnm>m mumz .vuo magma 117 Macrophyte control varied among ponds. In NW and SW ponds, 3 ppm copper sulfate completely controlled Potamogeton crispus, while Elodea Nuttallii, though severely damaged, was capable of regrowth after 6 to 8 weeks in SW pond. Removal of target species was temporary in SW and NW ponds. By the 17th day after treatment in SW pond blue- green periphyton covered most surfaces; growth of these mats continued until the 35th day. Lemna minor appeared by the 25th day, with Oscillatoria sp. and Hydrilla verticillata visible by the 28th day. Potamogeton crispus was noted by the 37th day. Within 90 days, macrophyte growth exceeded pre-treatment levels, and Oedogonium repopulation had begun. In NW pond, Euglena sp. and Phacus sp. populations developed by the 14th day after treatment and soon formed a green scum on the surface. Levels of P04 were determined in three of the systems (Figure C-l). Only in NE pond were PO4 levels studied before and after dosage; in this pond a peak PO concentration was 4 reached 11 days after dosage. Jewell (1971) noted that nutrient regeneration from decaying macrophytes reached a peak 5 to 10 days after the onset of decomposition. Readings of PO levels in NW pond, started 11 days after 4 dosage, reached a high of 1.45 ppm. Levels of P04 were higher in NW than in NE pond because plant mass and degree of decompo- sition was greater. In contrast, SE pond samples had no de- tectable P04. Levels reached in NW and NE ponds indicated an ample supply of phosphorus for algal blooms. Jewell (1971) 118 Figure C—l. Changes in P04 concentrations over time in NW, NE, and SE ponds. Shaded area denotes addition of 1 ppm copper sulfate to NE pond. 119 Northwest Pond Northeast Pond Southeast Pond 10-1 10-11 10-21 10-31 11-1 11-11 TIME Figure C-l 120 stated that since aquatic weeds decayed faster and more com- pletely than algae, they had greater potential for regenerat- ing nutrients. Appearance of blue-green periphyton in SW pond and Euglena and Phacus in NW pond indicated that planktonic algae present during nutrient regeneration bloomed, while the more slowly growing macrophytes reappeared later. In NW, SW, and NE ponds, a deterioration of aesthetic quality of water occurred as plant tissue decomposed. In NE pond, only slight odor and discoloration was noted. SW pond showed similar but somewhat more prolonged effects. In NW pond, where extensive 92252 decomposition occurred, the odor of H S was very offensive. The water became very dark, and 2 an oily scum of an organic nature appeared on the surface. As seen in Figure A—2, water chemistry changes in NW pond after ghapa decomposition were dramatic. Jewell (1971) noted that an indiscriminate application of herbicides to flowing or standing water might result in massive deoxygena- tion of the water. Owens and Maris (1971) applied paraquat to a small lake and noted complete deoxygenation in 4 days. Whitworth and Lane (1969) noted that when 5 ppm copper sulfate was added to a simulated natural pond, oxygen production was suppressed for 2 to 3 days. The effect in NW pond was dramatic with D.O. levels below 0.5 ppm for 10 days. Sudden drOps in pH and increases in hardness and methyl orange alkalinity also resulted from the release of decomposition products from plant tissues. 121 In both SW and NE ponds (Figures A-1 and A-3), changes in water chemistry similar to those noted in NW pond occurred but to a lesser degree. D.O. levels dropped rapidly but then rose quickly. Increases in methyl orange alkalinity and hard- ness and decreases in pH were less pronounced. The magnitude of change in chemical parameters was re- lated to degree of decomposition of plant tissues present. Warm water temperatures hastened decomposition and also may have affected the degree of change. Effects of drastic changes in water chemistry parameters on the biota were noted in NW pond (Part II). S UMMA RY An evaluation of the effectiveness of copper sulfate in the present study is difficult. Copper sulfate at 1 ppm con- trolled the filamentous alga Oedogonium sp. and Spirogyra sp., while at 3 ppm, control of Potamogeton crispus, Elodea Nuttal- lii, and Chara sp. was adequate. Control of planktonic algae species, Ceratium hirundinella and Trachelomonas sp., was-not achieved during the study. At 3 ppm copper sulfate doses, toxicity to some fish-food organisms, a deterioration of aesthetic quality, and the pos- sibility of drastic indirect effects from plant tissue decompo- sition existed. Extreme caution in using copper sulfate in small closed ponds is wise. During conditions of high tempera- tures and heavy algal or macrOphytic growth, drastic changes in the pond may result from copper sulfate dosage, even at low levels. Control may be temporary, as noted in both NW and SW ponds. 122 REFERENCES CITED 123 REFERENCES CITED American Public Health Association, gp‘gl. 1965. Standard Methods for the Examination of Water and Waste Water. 12th ed. New York, 873 pp. Anderson, R. O. 1959. A modified flotation technique for sorting bottom fauna samples. Limnol. Oceanogr. 4(2): 233-225. Arthur, J. W., and E. N. Leonard. 1970. Effects of copper on Gammarus pseudolimnaegs, Physa integra, and Campeloma dicIsum in soft water. J. Fish. Res. Bd. Canada 27: 1277-1283. Bartley, T. R. 1969. Copper residue on irrigation canal. Rep. Bur. of Reclam. 16 pp. Cairns, J., and A. Scheier. 1968. A comparison of the toxicity of some common industrial waste components tested individually and.Combined. Prog. Fish-Cult. 30(1):3-8. Chancellor, R. J., A. V. Coombs, and H. S. Foster. 1958. Control of aquatic weeds by copper sulphate. Proc. 4th Br. Weed Cont. Conf. 80-84. ' Cope, O. B. 1965. Some responses of freshwater fish to herbicides. Proc. South. Weed Conf. 18:439-445. Crance, J. H. 1963. The effects of copper sulfate of Microcystis and zooplankton in ponds. Prog. Fish-Cult. 25(4):198-202. Deubert, K. H., and I. E. Demoranville. 1970. Copper sulfate in flooded cranberry bogs. Pesticide Monitoring J. 4(1):ll-13. Eipper, A. W. 1959. Effects of five herbicides on farm pond plants and fish. J. New York Fish and Game 6:46-56. Environmental Protection Agency. 1971. Methods for Chemical Analysis of Water and Wastes. Environmental Protection Agency. Cincinnati, 312 pp. 124 125 Environmental Protection Agency. 1971. Water Quality Criteria Data Book. III. Effects of chemicals on aquatic life. Water Pollution Control Research Series. 100 pp. Fisher, A. 1956. The effect of copper sulfate on some micro- organisms in fish ponds. Bamidgeh 8(2):21-27. Fitzgerald, G. P. 1966. Use of potassium permanganate for control of problem algae. J. Amer. Water Wks. Assoc. 58(5):609-614. Gilderhus, P. A. 1966. Some effects of sublethal concentra- tions of sodium arsenite on bluegills and the aquatic environment. Trans. Amer. Fish. Soc. 95(3):289-296. Huckins, R. K. 1960. Aquatic weed control, Carnegie Lake, Princeton, New Jersey. Proc. 14th NE Weed Cont. Conf. 496—501. Hutchinson, G. E. 1957. A Treatise on Limnology. John Wiley & Sons, Inc., New York. 1015 pp. Jewell, W. J. 1971. Aquatic weed decay: dissolved oxygen utilization and nitrogen and phosphorus regeneration. J. Water Poll. Contr. Fed. 43(7):l457-l467. Kariya, T., Y. Haga, H. Haga, and T. Tsuda. 1967. Studies on the post-mortem identification of the pollutant in the fish killed by water pollution. V. Detection of copper in the fish. Bull. Jap. Soc. Sci. Fish. 33(9): 818-824. Kopp, J. F., and R. C. Kroner. 1967. A five year summary of trace metals in rivers and lakes of the United States. U. S. Dept. of the Interior; Compilation of Data, October 1, l962-—September 30, 1967. 28 pp. Lucas, H. F., Jr., D. N. Edgington, and P. J. Colby. 1970. Concentrations of trace elements in Great Lakes fishes. J. Fish. Res. Bd. Canada 27(4):677-684. Lucas, R. E. 1948. Chemical and physical behavior of copper in organic soils. Soil Sci. 66:119-129. Mackenthun, K. M., and H. L. Cooley. 1952. The biological effect of copper sulphate treatment on lake ecology. Trans. Wisc. Acad. Sci., Arts, and Letters 41:177-187. McKim, J. M., and D. A. Benoit. 1971. Effects of long-term exposures to copper on survival, growth, and reproduction of brook trout (Salvelinus fontinalis). J. Fish. Res. Bd. Canada 28(5):655-662. 126 McNabb, C. D. 1960. Enumeration of freshwater phytoplankton concentrated on the membrane filter. Limnol. Oceanogr. 5(1):57-6l. Mount, D. I. 1968. Chronic toxicity of copper to fathead minnows (Pimephales promelas, Rafinesque). Water Res. 2:215-223. Mulligan, H. F. 1969. Management of aquatic vascular plants and algae. In Eutrophication: Causes, Consequences, Correctives. National Academy of Science, Washington, D. C. 661 pp. Nelson, J. L., V. F. Bruns, C. C. Coutant, and B. C. Carlile. 1969. Behavior and reactions of copper sulfate in an irrigation canal. Pesticide Monitoring J. 3(3):186-189. Nichols, M. S., T. Henkel, and D. McNall. 1946. Copper in lake muds from lakes of the Madison area. Trans. Wisc. Acad. Sci., Arts, and Letters 38:333-350. O'Hara, J. 1971. Alterations in oxygen consumption by blue- gills exposed to sublethal treatment with copper. Water Res. 5:321-327. Ong, H. L., V. E. Swanson, and R. E. Bisque. 1970. Natural organic acids as agents of chemical weathering. U. S. Geol. Survey Prof. Paper 700-C: C-l30-C-l37. Otto, N. E. 1970. Algaecidal evaluation and environmental study of mat producing blue-green algae. Bur. Reclam. Owens, M., and P. J. Maris. Some ecological effects of the use of paraquat for weed control on a small lake. II.’ Effects of aquatic weed control on the productivity and some chemical characteristics of a small lake. J. Appl. Ecol. (In press). Pennak, R. W. 1953. Freshwater Invertebrates of the United States. Ronald Press Co., New York. 769 pp. Phelps Dodge Refining Corporation. n.d. Controlling plant and animal pests in farm ponds with copper sulfate. 32 pp. Pickering, O. H., and C. Henderson. 1965. The acute toxicity of some heavy metals to different species of warm water fishes. Proc. 19th Ind. Waste Conf. 49(2):578-591. Prescott, G. W. 1964. How to Know the Fresh-Water Algae. Wm. C. Brown Co., Dubuque. 272 pp. 127 Riemer, D. N., and S. J. Toth. 1970. Adsorption of c0pper by clay minerals, humic acid and bottom muds. J. Amer. Water Wks. Assoc. 62(3):l95-197. 1968. A survey of the chemical composition of aquatic plants in New Jersey. New Jersey Agri. Exp. Stat. Tech. Bull. 820. 14 pp. Riley, G. A. 1939. Limnological studies in Connecticut. Ecol. Monogr. 9:54-94. Rose, E. T. 1954. Blue green algae control at Storm Lake. Proc. Iowa Acad. Sci. 61:604-614. Smith, M. W. 1939. Copper sulphate and rotenone as fish poisons. Trans. Amer. Fish. Soc. 69:141-157. Sohacki, L. P. 1965. Ecological alterations produced by the treatment of pond ecosystems with copper sulfate and sodium arsenite. M. S. Thesis. Michigan State Univer- sity. 91 pp. Stiff, M. J. 1971. The chemical states of copper in polluted fresh water and a scheme of analysis to differentiate them. Water Res. 5:585-599. Surber, E. W., and M. H. Everhart. 1950. Biological effects of Nigrosine used for control of weeds in hatchery ponds. Prog. Fish-Cult. 12:135-140. Sutton, D. L., and R. D. Blackburn. 1971. Uptake of copper in Hydrilla. Weed Res. ll(l):47-53. , L. W. Weldon, and R. D. Blackburn. 1970. Effect of diquat on uptake of copper in aquatic plants. Weed Sci. 18(6):703-707. Tobia, S. K., and A. S. Hanna. 1958. Effects of copper sul- fate added to irrigation water on copper status of Egyptian soils. I. Amount of c0pper retained by soils. Soil Sci. 85:302-306. Tompkins, W. A., and C. Bridges. 1958. The use of copper sulfate to increase fyke-net catches. Prog. Fish-Cult. 20(1):16-20. Toth, S. J., and D. N. Riemer. 1968. Precise chemical con- trol of algae in ponds. J. Amer. Water Wks. Assoc. 60(3):367-37l. 128 Walker, C. R. 1961. Toxicological effects of several herbi- cides to bottom dwelling fish-food organisms in Missouri ponds. Res. Rept. 18th NC Weed Cont. Conf.: 104. Ware, F. J. 1966. The use of copper sulfate as a method of partial control of elodea (Elodea densa) in Lake Thonotosassa, Florida. Proc. 19th Sth. Weed Conf.: 491-494. Warnick, S. L., and H. L. Bell. 1969. The acute toxicity of some heavy metals to different species of aquatic insects. J. Water Poll. Contr. Fed. 4(2):280-284. Whitworth, W. R., and T. H. Lane. 1969. Effects of toxi- cants on community metabolism in pools. Limnol. Oceanogr. l4(l):53-58. APPENDIX 129 130 Table D-l. Per cent recovery of copper from spiked samples. Material Per cent recovery Plant Tissue: Elodea Nuttallii 92.3 Elodea Nuttallii 106.4 Elodea Nuttallii 103.5 Elodea Nuttallii 92.9 Elodea Nuttallii 92.4 Chara sp. 100.6 Chara sp. 107.9 Chara sp. 99.7 Potamogeton crispus 108.1 Potamogeton crispus 114.4 Potamogeton crispus 104.3 Oedogonium sp. _’ 89.9 x = 101.2 Sediment: 1. 120.0 2. 112.1 3. 108.5 4. 102.6 5. 94.7 6. 116.3 7. 108.7 3'5 = 109.0 Table D-2. 131 Reproducibility of sample readings. Sample number Reading 1 (mg Cu) Reading 2 (mg Cu) 1 0.266 0.4728 2 2.098 2.01 3 0.765 0.648 4 0.702 0.712 5 0.498 0.576 6 0.416 0.406 7 0.374 0.368 8 0.646 0.676 9 0.492 0.494 10 0.096 0.106 11 1.180 1.156 Note: when tested with a paired t-test (2-tailed), the 2 sets of means were not significantly different at the 5% level. 132 Table D-3. Green sunfish stomach analyses. Weight Date Content (gm) SW Pond 3.829 6-21 Several Chaoborus; culicids; chironomids 2.878 6-21 Several mayflies and cladocera; l culicid 3.971 6-28 1 mayfly; l dytiscid; 25 cladocera 8.067 7-19 2 c0pepods 5.430 7-19 65 copepods; 1 damselfly 12.952 7-19 25 copepods; 10 chironomids; 1 caddisfly; l mayfly 5.57 7-19 4 chironomids; l aquatic mite 9.758 7-19 3 caddisflies; 6 chironomids 6.329 7-20 70 copepods; 2 chironomids 7.592 7-20 Several copepods 6.925 7-20 4-5 mayflies 7.465 8-3 30 cladocera; 10 ostracods; 8O copepods 6.58 8-3 Several copepods 5.292 8-3 4 ostracods 9.135 8-15 14 chironomids; l ostracod; 100 cladocera 29.216 11-16 1 dragonfly naiad NE Pond 7.523 6-25 Several copepods 5.716 7-1 1 ostracod; 3 cladocera; 1 chironomid; 160 copepods 2.929 6-25 Several oligochaetes 2.017 6-25 20 copepods 9.036 8-5 Several ostracods; several mayflies 7.214 8-19 Several copepods; several chironomids 7.159 8-19 Several c0pepods; l mayfly; 2 chironomids 11.456 8-19 Parts of mayflies continued Table D-3--continued 133 Weight Date Content (gm) 12.154 10-15 1 mayfly 10-26 9 chironomids 10-26 6-8 chironomids 10-26 1 mayfly NW Pond 5.225 6-21 Several mayflies; 1 chironomid 4.235 6-21 Several mayflies and several chironomids 3.964 6—28 1 mayfly; 5 chironomids; 10 cladocera; 3 dytiscids 4.579 7—20 30 cladocera; l dytiscid; 1 coleopteran 11.339 7-20 Several cladocera; 1 ostracod; 2 caddis- flies; several chironomids; l mayfly; l dytiscid 7.428 8-28 2 copepods; 2 snails; l ostracod 6.135 8-28 Empty 9.823 9—30 15 dytiscid larva; several haliplid larva 11.619 10-7 9 chironomids 7.121 10-7 7 chironomids; 3 copepods 9.774 10-7 Empty 7.269 10-7 Several copepods 10.969 10-7 3 chironomids SE Pond 4.072 6—25 Several copepods 2.772 6-25 130 c0pepods 3.657 6-25 Several copepods; l aquatic oligochaete 7.342 8-5 20 copepods; l chironomid 9.426 8-5 1 snail; several copepods; 1 cladocera; 1 chironomid continued Table D-3--continued 134 Weight Date Content (gm) 10.106 8-5 5 mayflies; l copepod; l cladoceran 8.830 8-19 Empty 10.202 8-19 Empty 8.306 8-19 Empty 9.982 8-22 90 copepods; 25 chironomids 7.287 8-22 5 chironomids 9.600 8-22 300 cladocera; 10 copepods; several ostracods 11.114 8—26 110 copepods; several chironomids 8.816 8-26 Empty 9.683 8-26 3 snails; l chironomid; 350 c0pepods 9.171 8-26 Several copepods 11.614 9-30 8 copepods; several ostracods 13.147 9-30 15 copepods HICHIGRN STATE UNIV. 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