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HomeMy WebLinkAboutAPA2647Effects on Arctic Grayling (Thymallus arctlcus) of Short-Term . . Exposure to Yukon Placer Mining Sediments: Laboratory ·and Field Studies D. J. Mcleay, A. J. Knox~ J. G. Malick, I. K. Birtwell, G. Hartman, and G. L. -Ennis :Department of Fisheries and Oceans Fisheries Research Branch · West Vancouver Laboratory 4160 Marine Drive West Vancouver, British Columbia V7V 1 N6 • 0 May 1983 Canadian Technical Report of Fisheries and Aquatic Sciences N.o. 1171 ment of Canada Gouvernement du Canada s and Oceans Peches et Oceans Canadian Technical Report of Fisheries and Aquatic Sciences Technical reports contain scientific and technical information that contributes to existing knowledge but which is not normally appropriate for primary literature. Technical reports are directed primarily toward a worldwide audience and have an international distribution. 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Les rapports epuises seront fournis contre retribution par des agents commerciaux. Canadian Technical Report of Fisheries and Aquatic Sciences No. 1171 Yukon River Basin Study Fisheries Project Report No. 2 May 1983 EFFECTS ON ARCTIC GRAYLING (Thymallus arcticus) OF SHORT-TERM EXPOSURE TO YUKON PLACER HINING SEDIMENTS: LABORATORY AND FIELD STUDIES by D.J. McLeay 1 , A.J. Knox 2 , J.G. Malick2 , I.K. Birtwell3a, G. Hartman3b and G.L. Ennis3c 1 n. McLeay & Associates Ltd., Suite 300, 1497 Marine Drive, West Vancouver, B. C. V7T 1B8 2 Norecol Environmental Consultants Ltd., Suite 100, 1281 West Georgia St., Vancouver, B. C. V6E 3J7 3 Fisheries and Oceans Canada a) Salmon Habitat Section, Fisheries Research Branch, West Vancouver Laboratory, 4160 Marine Drive, West Vancouver, B. C. V7V 1N6 b) Salmon Habitat Section, Fisheries Research Branch, Pacific Biological Station, Nanaimo, B. C. V9R 2Pl c) Habitat Management Division, Field Services Branch, 1090 West Pender Street, Vancouver, B. C. V6E 2Pl -11 - (c) Minister of Supply and Services Canada 1983 Cat. No. Fs 97-6/1171 ISSN 0706-6457 [ [ c [ c c c ...c •c c f" L [ [ -iii - PREFACE These studies,with other preliminary work on food habits and the distribu- tion of Arctic grayling,were carried out to provide some initial information ~n the effects of placer mining sediments on this species of fish.At the time the 1982 field work was plan~ed,it was anticipated that a longer term programme of more comprehens ive studies on the effects of sediment on various stages in the life cycle of grayling and their habitat would follow.While this work was directed primarily at sediment impacts on juvenile grayling,it was also recognized that a sound understanding of the ecology of the species is needed for management purposes. Sensitive and well informed management of water resources,and the associated protection of fisheries values,will require much more research on the effects of sediment impacts on a wide variety of biological processes.Such research will require elucidation of the effects of,for example,the concentration of sediment, size,shape and hardness of particles,mixes of particle types and timing of discharge.Studies on many of these factors should be carried out on fish at various stages in their life cycle and in different seasons.In addition,studies on the effects of sediment and sediment characteristics on rheotactic behaviour, feeding behaviour,spawning behaviour,and the production of food organisms for fish;are required to support sound water use planning.Although it may be desirable to have information on this scale now for water resource decisions,it is not realistic to expect it after one year of research. It is hoped that the work done in 1982,and considered in total,may be of value to the agencies that manage water resources for the public of Yukon. However,the authors would warn people not to use single components of the results to form guiding principles in water use decision making.For example, acute lethal bioassay tests,performed in otherwise protected conditions,do not in themselves indicate the effect of much lower concentrations of sediment on grayling in the wild which must find food,avoid predators,and maintain positions in a stream system over a prolonged period.Other species of fish have been shown to be able to tolerate exposure,in protected conditions,to short-term high concentrations of sediment.It has also been shown that the same species are affected adversely by much lower concentrations of sediment where physiological tests are considered or where reproduction and feeding are involved. Part.s of the work we carried out in 1982 indicate the nature of certain physiological responses of fish to suspended sediment.Other components investigated the distribution and food of grayling in a stream system receiving sediment from placer mining operations;the results of this work will be reported separately. In this Preface the authors are not apologizing for the scale or quality of these initial studies.We are urging caution in interpretation and application of such first-stage research.In a broad sense we are stressing the need to understand cold-zone stream ecology,grayling biology,and the complex effects of various components of placer mining on them. The Department of Fisheries and Oceans (Fisheries Research Branch and Field Services Branch)and the Yukon River Basin Study (a joint study by Canada,Yukon and British Columbia of the water and related resources of the Yukon basin)funded this project.Opinions expressed are those of the authors.The work was a co-operative undertaking by D.McLeay &Associates Ltd.,Noreco1 Environmental Consultants Ltd.,and staff of the Department of Fisheries and Oceans.The study also relied upon the co-operation of placer miners along Highet Creek,within the Minto Creek drainage. iv TABLE OF CONTENTS [ '[ List of Tables .• List of Figures ••••...............................'. vi ix [ 1S Q C. SaC •. tests. 1S o C. List of Appendices . Abs tr ac t IRe-sume . Introduction ..••• Materials and Methods. Laboratory Studies .. Fish collection ... Fish rearing .•••.• Sediment collection...••.•.••••••• Sediment preparation and analyses .. Recycle test tanks •••. Acute survival tests •. Fish acclimated to Fish acclimated to Temperature tolerance Fish acclimated to Fish acclimated to SoC .• Sealed jar bioassays .•.••••• Fish acclimated to 15°C •.•••••• Fish acclimated to SaC. Acute stress bioassays ••.. Reference toxicant tests •• Statistical analyses. Field Studies ••. Study area •••••••. General . Highet Creek .• Minto Creek •.. Fish collection •. In~situ bioassays .. Test apparatus .. Water quality •• Experimental ••• Results and Discussion. Laboratory Studies •.•.....••••.•.•. Fish growth and condition •• Characteristics of test sediment ••• Acute survival and gill histology •• Temperature tolerance tests. Sealed jar bioassays •.• Acute stress bioassays. General ...........••.•• xii xiii 1 2 2 2 2 3 3 S S S 7 7 7 8 8 8 9 9 10 10 11 11 11 12 13 13 14 14 14 IS 16 16 16 18 20 22 24 26 28 [ c c ., [ [ [ [ [ [ [ [' - v - TABLE OF CONTENTS (CONT.) ·. Field Studies •.•••• Water quality .•. Caged fish studies. Fish survival.. Gill histology. Hematology .••••• [ [ [ [ [ o c • [ [ [ [ [, [ Conclusions •••••. Acknowledgements. References .•.•..• ·. ·. 29 29 32 32 32 34 34 35 36 Table 1 2 3 4 S 6 7 8 9 10 11 12 -vi - LIST OF TABLES Particle size distribution for paydirt and overburden sediment samples. Moisture content,volatile and fixed residue,and oxygen uptake rate for paydirt and overburden sediment samples. Metal content of paydirt and overburden sediment samples. Effect of location within recycle test tanks on concen- tration (total residue values)of recirculating paydirt sediment. Acute survival test:Effect of a 4-day exposure to sus- pended inorganic paydirt fines on fish survival and on blood sugar and hematocrit values for underyear1ing Arctic grayling acclimated to lSoC. Acute survival test:Effect of a 4-day exposure to suspended organic overburden on fish survival and on blood sugar and hematocrit values for underyear1ing Arctic grayling acclimated to lSoC. Acute survival test:Effect of a 4-day exposure to sus- pended inorganic paydirt fines on fish survival and on blood sugar,1eucocrit and hematocrit values for under- yearling Arctic grayling acclimated to SoC. Temperature tolerance test:Effect of suspended inorganic paydirt"on the critical thermal maxima forunderyear1ing Arctic grayling acclimated to lSoC. Temperature tolerance test:Effect of suspended organic overburden on the critical thermal maxima for underyear- ling Arctic grayling acclimated to lSoC. Temperature tolerance test:Effect of pentachlorophenol on the critical thermal maxima for underyear1ing Arctic grayling acclimated to lSoC. Temperature tolerance test:Effect of suspended inorganic paydirt on the critical thermal maxima for underyear1ing Arctic grayling acclimated to SoC. Sealed jar bioassay:Effect of suspended inorganic pay- dirt on tolerance to hypoxia and time to hypoxic death for underyear1ing Arctic grayling acclimated to lSoC. [ r L l~ [ [ [ [ [ .. [ [ [ [ [ [ -[ f" '''-. L [ [ [ [ [ c ....c o [ c [ [ [. E· [ Table 13 14 15 16 17 18 19 20 21 -vii - LIST OF TABLES (CONT.) Sealed jar bioassay:Effect of suspended inorganic paydirt on tolerance to hypoxia and time to hypoxic death for underyearling Arctic grayling acclimated to 5°C. Sealed jar bioassay:Effect of suspended organic overburden on tolerance to hypoxia and time to hypoxic death for underyearling Arctic grayling acclimated to 15°C. Sealed jar bioassay:Effect of pentachlorophenol on tolerance to hypoxia and time to hypoxic death for underyearling Arctic grayling acclimated to 15°C. Acute stress bioassay:Effect of suspended inorganic paydirt on blood sugar,hematocrit and leucocrit values for underyearling Arctic grayling acclimated to 15°C. Acute stress bioassay:Effect of suspended organic overburden on blood sugar,hematocrit and leucocrit values for underyearling Arctic grayling acclimated to 15°C. Acute stress bioassay:Effect of pentachlorophenol on blood sugar,hematocrit and leucocrit values for °underyearling Arctic grayling acclimated to 15 C. Summary of threshold-effect concentrations of paydirt or overburden suspensions causing acute responses for Arctic grayling. Water quality characteristics monitored at test site in Highet Creek and the control site in Minto Creek during the fish enclosure tests,August and September, 1982. Hardness,alkalinity and metal content (mgoL-1 ) determined for water samples taken from Highet,Minto and Mud creeks during the fish enclosure tests, August and September,1982. Table 22 23 24 25 -viii - LIST OF TABLES (CONT.) Particle size distribution for suspended sediment sampled from Highet Creek during August and September,1982. Percentage survival of underyearling Arctic gray- ling held in Highet Creek or Minto Creek for 4-5 days during August or September,1982. Gill histopathologies for underyearling Arctic grayling held in Minto Creek or Highet Creek during September 1982. Mean (±SD)biological characteristics of underyear- ling Arctic grayling sampled from cages or directly from creeks during August and September,1982. [ .r L [ [ [ [ c .... [ [ [ [ [ l [ [>- ['I / [ [ [ c c 0 ... 0 c c [ c [ [ [< [. [ / -ix - LIST OF FIGURES Figure 1 Illustration of recycle test tanks. 2 Illustration of derivation of hematocrit and leucocrit values from a centrifuged blood sample within a heparin- ized glass capillary tube. 3 Map of site for in-situ caged fish studies. 4 Schematic drawing of net enclosures for in-situ caged fish studies. S Study site at Minto Creek.Fish enclosures are shown in-situ. 6 Study site at Highet Creek.Fish enclosures are shown in-situ. 7 Relationship of total residue,nonfiltrable residue and turbidity for suspensions of paydirt sediment in fresh- water. 8 Relationship of total residue,nonfiltrable residue and turbidity for suspensions of overburden sediment in freshwater. 9 Illustration of the stability of differing concentrations of suspended paydirt fines within recycle test tanks during a 96-h bioassay. 10 Relationship of concentration of suspended inorganic pay- dirt to critical thermal maxima for underyearling Arctic grayling acclimated to lSoC. 11 Relationship of concentration of suspended organic over- burden to critical thermal maxima for underyearling Arctic grayling acclimated to lSoC. 12 Relationship of concentration of suspended inorganic pay- dirt to critical thermal maxima for underyearling Arctic grayling acclimated to SoC. 13 Relationship of concentration of inorganic paydirt to time to death in sealed jar bioassays for underyearling Arctic grayling acclimated to lSoC and tested at 20°C. Figure 14 lS 16 17 18 19 20 21 22 - x - LIST OF FIGURES (CaNT.) Relationship of concentration of inorganic paydirt to tolerance to hypoxia in sealed jar bioassays for underyear1ing Arctic grayling acclimated to lSoC and tested at 20°C. Relationship of concentration of inorganic paydirt to time to death in sealed jar bioassays for underyear- ling Arctic grayling acclimated to SoC and tested at 10°C. Relationship of concentration of inorganic paydirt to tolerance to hypoxia in sealed jar bioassays for underyear1ing Arctic grayling acclimated to SoC and tested at 10°C. Relationship of concentration of organic overburden to time to death in sealed jar bioassays for under- yearling Arctic grayling acclimated to lSoC and tested at 20°C. Relationship of concentration of organic overburden to tolerance to hypoxia in sealed jar bioassays for underyear1ing Arctic grayling acclimated to lSoC and tested at 20°C. Relationship of concentration of suspended inorganic paydirt to blood 1eucocrit values for underyear1ing Arctic grayling acclimated to lSoC and exposed to sediment for 24 h. Relationship of concentration of suspended organic overburden to blood 1eucocrit values for underyear- ling Arctic grayling acclimated to lSoC and exposed to sediment for 24 h. Relationship of concentration of suspended inorganic paydirt to blood sugar values for underyear1ing Arctic grayling acclimated to lSoC and exposed to sediment for 24 h. Relationship of concentration of suspended organic overburden to blood sugar values for underyear1ing Arctic grayling acclimated to lSoC and exposed to sediment for 24 h. r rL~ r [ [ [ [ [ [ c [ [ [ [ [ ,[ [ [ [ [ [ [ [ c [ [ [ [ [. Figure 23 24 25 26 27. -xi - LIST OF FIGURES (CONT.) Relationship of concentration of suspended inorganic paydirt to blood sugar values for underyearling Arctic grayling acclimated to 5°C and exposed to sediment for 96 h. Illustration of concentration of suspended sediment (total residue)and turbidity within cages held in Highet Creek during August 1982. Illustration of concentration of suspended sediment (total residue)and turbidity within cages held in Highet Creek during September 1982. Gill filaments of underyearling Arctic grayling·captured from Minto Creek during September 1982.Note normal appearance of secondary lamellae (a).300X. Gill filaments of underyearling Arctic grayling captured from Minto Creek and held in a cage within Minto Creek for 5 days during September 1982.Note moderate hypertrophy (increase in cell size)and hyperplasia (increase in cell numl;>ers .of lamellar epithelium (b),and presence of large numbers of ectoparasites (c).300X. Appendix 1 2 3 4 S 6 7 8 9 -xii - LIST OF APPENDICES Summary of the aquatic biophysical characteristics for the Highet Creek and Minto Creek caged fish sites during August and September 1982. Physical/chemical characteristics during 4-day survival test with lSoC-acc1imated underyear1ing Arctic grayling exposed to inorganic paydirt suspensions. Physical/chemical characteristics during 4-day survival test with lSoC-acc1imated underyear1ing Arctic grayling exposed to organic overburden suspensions. Physical/chemical characteristics during4-day survival test with SOC-acclimated underyear1ingArctic grayling exposed to inorganic paydirt suspensions. Residue and turbidity values within a cage held in Highet Creek during the August 1982 in-situ fish survival test. Residue and turbidity values within a cage held in Highet Creek during the September 1982 in-situ fish survival test. Residue and turbidity values within a cage held in Minto Creek during the August 1982 in-situ fish survival test. Residue and turbidity values within a cage held in Minto Creek during the September 1982 in-situ fish survival tes. Comparison of suspended sediment and turbidity values for triplicate water samples taken from within or out- side of a Highet Creek cage during the August and September 1982 in-situ fish survival tests. [ rL [ [ [ [ c c [ [ [ .[ L [ [' [ [ [ c o C c c [ c· [ -xiii - ABSTRACT McLeay,D.J.,A.J.Knox,J.G.Malick,I.K.Birtwell,G.Hartman and G.L.Ennis. 1983.Effects on Arctic grayling (Thymallus arcticus)of short-term exposure to Yukon placer mining sediments:laboratory and field studies.Can.Tech.Rep. Fish.Aquatic Sci.1171:xvii +134 p ° A program of controlled laboratory and in-situ field bioassays was conducted during 1982/83 to examine the acute effects Of·suspensions of Yukon placer mining sediment on underyearling Arctic grayling (Thymallus arcticus).wild grayling, captured as swimup fry or young fingerlings,were acclimated to warmwater (lSOC) or coldwater (SOC)conditions for 7-12 weeks,and subjected to a range of concentrations of organic sediment (overburden)and/or inorganic sediment (paydirt)suspensions in recirculating test tanks.On two occasions (August and September 1982),grayling fingerlings were captured from central Yukon clearwater streams and held for 4 or S days in cages within turbid creekwater (Highet Creek) downstream of placer mining activities,and at a nearby clearwater site (Minto Creek upstream of its junction with Highet Creek). Laboratory-reared grayling acclimated to lSoC survived a 4-day exposure to inorganic sediment suspensions <2S0,000 mgoL-l,and a l6-day exposure to SO,OOO mg °L -1.These fish also survived acute (4-day)exposure to all strengths of organic sediment examined «SO,OOO mgoL-l).All fish acclimated to SoC and held in paydirt suspensions <10,000 mgoL-l survived for 4 days,whereas 10-20% mortalities occurred in the higher strengths examined. Inorganic sediment strengths >10,000 mgoL-l caused fish to surface,a direct response to elevated sediment levels.No other behavioural anomalies were evident.Other signs of fish distress or damage were not observed for any grayling surviving exposure to either sediment type.The gill histology of fish surviving these 4-day exposures was normal. The tolerance of laboratory-reared grayling to temperature extremes (critical thermal maxima)was not impaired appreciably by either sediment type.Slight but consistent declines in critical thermal maxima were noted for warmwater-acclimated fish held in inorganic or organic sediment strengths >SOO mg °L -1 and >S ,000 mgoL-l,respectively,whereas changes in thermal tolerance-were not found for fish acclimated to cold water and held in high strengths of inorganic sediment. The acute tolerance of warmwater-or coldwater-acclimated fish to hypoxic conditions (oxygen deficiency)in sealed jar bioassays was not impaired by suspended sediment.Tests with overburden suspensions showed a decreased time to death in these bioassays,which was attributed to the sediment I s oxygen demand. High concentrations of paydirt increased time to death (decreased respiratory rate)in sealed jar bioassays for the warmwater-acclimated fish only. Suspensions of inorganic and organic sediment caused acute stress responses (elevated and/or more varied blood sugar levels,depressed leucocrit levels)for grayling acclimated to either temperature.Responses were noted for sediment -xiv -[ strengths as low as 50 mgoL-l (overburden),although confirmation of threshold- effect levels requires further studies.Hematocrit values for these fish were not affected by sediment. Acute (short-term)effects toward Arctic grayling of the reference toxicant pentachlorophenol were examined in laboratory biossays.Median lethal concentrations were similar to those found previously with this aquatic contaminant and other species of salmonid fish,and were not affected by acclimation temperatures.The effects on grayling of sublethal strengths of pentachlorophenol noted for temperature tolerance tests,sealed jar bioassays and acute stress bioassays were also similar to those determined before with other juvenile salmonids. During the August field bioassays,all grayling held in Highet Creek (suspended solids <l00 mgoL-l)or Minto Creek (suspended solids <20 mgoL-l)for 4 days survived,with no overt signs of distress or physical damage.In September, all fish captured from Minto Creek and held in cages within Highet Creek (suspended solids <1,210 mgoL-1 )or Minto Creek (suspended solids <34 mgoL-1 )for 5 d~ys also survived.\Gill tissues of fish sampled in September from cages at each site showed moderate-to-marked hypertrophy and hyperplasia of lamellar epithelium,together with a proliferative number of gill ectoparasites.No histopathological differences were found between sites.The gill histology of uncaged grayl ing sampled directly from Minto Creek upstream of Highet Creek was kormal,although occasional ectoparasites were observed. I All grayling captured from Mud Creek (a clearwater tributary of Minto Creek) and held for the same 5-day period during September in cages within Minto Creek survived;whereas 16%(5 {'4}f 32 fish)of the Mud Creek fish held at this time in Highet Creek',died wi thiriJ 96 h.The cause of these deaths was attributed to an intolerable stess loadin imposed by the combined effects of fish capture, transport,confinement and exposure to suspended sediment and temperaure fluctuations'within Highe Creek. Although hematocri values measured for fish caged at either site were I similar,mean plasma g\ilcose values for fish held for 4 days within Highet Creek during August were ele~~ted 30%from values for fish caged in Minto Creek at this time.During SeptemD~r,grayling captured from either Minto Creek or Mud Creek and caged in Highet Creek showed a 100%increase in mean plasma glucose levels, relative to values for corresponding groups held in Minto Creek.These differences were thought to be caused by the more stressful water quality conditions (suspended sediment loadings and/or more extreme temperature differences)within Highet Creek,compared with the Minto Creek site. It was concluded that the short-term exposure of Arctic grayling to sublethal concentrations of suspended inorganic or organic sediment can cause a number of effects including acute stress responses.In light of these findings,the environmental impact of placer mining sediments on the immediate and long-term adaptive capabilities (including feeding and other behavioural responses,disease resistance,growth and chronic well-being)of this sensitive fish species needs to be more fully understood. [ r [ [ [ [ c c t .- [ c [ [ [ [ l [ [, [' [J [ [ [ c o C ~'l C c c c [ [ [. [ -xv - McLeay,D.J.,A.J.Knox,J.G.Malick,I.K.Birtwell,G.Hartman,and G.L.Ennis.1983.Effects on Arctic grayling (Thymallus arcticus of short-term e4xposure to Yukon placer mining sediments:Laboratory and field studies.Can.Tech.Rep.Fish.Aquat.Sci.1171:xvii +134 p. Le present rapport porte sur un programme de bio-essais contrOles en laboratoire et sur Ie terrain menes en 1982-1983 afin d'etudier les effets aigus de sediments en suspension provenant de l'exploitation de gisements alluvionnaires au Yukon sur des individus de mains d'un an d'ombre arctique (Thymallus arcticus).Des ombres sauvages,capturees au stade d'alevins nageurs ou jeunes digitales,ont ete acclimatees a l'eau chaude (1S0C)ou froide (5°C)pendant 7 a 12 semaines et soumises a une gamme de concentrations de sediments organiques (morts-terrains de recouvrement)et inorganiques (riches graviers aurifares)en suspension dans des bassins d'essai a renvoi. Des digitales ont ete prises dans des ruisseaux d'eau claire du Yukon central, soit en aoOt et en septembre,et maintenues pendant 4 ou 5 jours dans des cages placees dans l'eau turbide du ruisseau Highet en aval de l'exploitation miniare alluvionnaire et dans l'eau claire du ruisseau Minto,en amont de sa jonction avec Ie ruisseau Highet. Les ombres elevees en laboratoire et acclimatees a 1Soc ont survecu a une exposition de 4 jours a des sediments organiques en suspension <2S0 000 mg.L-1 et de 16 jours a 50 000 mg.L-1.Ces poissons ont aussi survecu a une exposition aigue (4 jours)a toutes les concentrations de sediments organiques testees «SO 000 mg L-1).Tous les individus acclimates a soC et gardes dans des suspensions d'alluvions exploitables <10 000 mg.L-1 ont survecu pendant 4 jours alors que la mortalite variait de 10 a 12 %en presence de concentrations plus elevees. Des concentrations de sediments inorganiques >10 000 mg.L-1 forcaient les poissons a faire surface,ce qui represente une reaction directe a des niveaux eleves de sediments.Aucune autre anomalie de comportement n'a ete notee.Aucun signe d'epuisement ou de dommage n'a ete remarque chez les ombres qui ont survecu a une exposition aux deux types de sediments.Les preparations histologiques des ouies de poissons apras 4 jours d'exposition ne revelaient aucune anomalie. La tolerance des ombres elevees en laboratoire aux extr~mes thermiques (maximums thermiques critiques)n'a pas ete grandement diminuee par les deux types de sediments.Des baisses faibles mais constantes des maximums thermiques critiques ont ete notees chez les poissons acclimates a l'eau chaude et maintenus dans des concentrations de sediments organiques et inorganiques >SOO mg.L-1 et > S 000 mg.L-1 respeetivement elors que des variations de-Ia tolerance thermique n'ont pas ete decouvertes chez les poissons acclimates a l'eau froide et maintenus dans des concentrations elevees de sediments inorganiques. -xvi - La tolerance aigue des poissons acclimates a l'eau chaude et froide aux conditions hypoxiques (carence d'oxygene)pendant des bio-essais menes dans des bocaux scelles n'a pas ete reduite par les sediments en suspension. En presence de suspensions de morts-terrains de recouvrement,la mort etait plus rapide a cause de la demande en oxygene des sediments.Des concentrations elevees d'alluvions exploitables retardaient la mort (taux de respiration abaisse)des poissons acclimates a l'eau chaude seulement. Des suspensions de sediments inorganiques et organiques causaient des reactions de stress aigu (niveaux de sucre sanquin eleves au plus variables et niveaux abaisses de leucocrites)chez-les ombres acclimatees aux deux temperatures.Les reactions aux concentrations de sediments aussi faibles que 50 mg.L-1 (morts-terrains de recouvrement)ant ete notees quai que une confirmation des niveaux de seuil requiere des etudes plus poussees.Les valeurs de l'hematocrite chez ces poissons n'etaient pas affectees par la presence de sediments. On a aussi etudie l'incidence aigue (8 court terme)d'une substance toxique etalon,Ie pentachlorophenol,sur l'ombre arctique au cours de bio-essais en laboratoire.Les concentrations letales medianes de ce contaminant aquatique,semblables a celles notees precedemment chez d'autres especes de salmonides,n'etaient pas touchees par les temperatures d'acclimatation.Les effets de concentrations subletales de pentachlorophenol sur les ombres,notes pendant des tests de tolerance thermique,des bio-essais en bocaux scelles et des analyses biologiques du stress aigu,etaient semblables a ceux determines auparavant chez d'autres salmonides juveniles. Pendant les bio-essais sur Ie terrain menes en aoOt,toutes les ombres gardees dans Ie ruisseau Highet (solides en suspension <100 mg.L-1)et Ie ruisseau Minto (solides en suspension <20 mg.L-1)pendant 4-jours ant survecu sans signes evidents d'epuisement-ou de dommage physique.En septembre,taus les poissons captures dans Ie ruisseau Minto et maintenus dans des cages dans Ie ruisseau Highet (solides en suspension <1 210 mg.L-1)au Ie ruisseau Minto (solides en suspension <34 mg.L-1)pendant-5 jours avaient aussi survecu.Des echantillons d'ouies de poissons recueillis en septembre dans les cages de chaque site indiquaient une hypertrophie variant de moderee a marquee et une hyperplasie de l'epithelium lamellaire,en plus d'une proliferation numerique des ectoparasites des ouies.II n'y avait aucune difference histopathologique entre les deux endroits.Les preparations histologiques d'ouies d'ombres en liberte capturees dans Ie ruisseau Minto en amant du ruisseau Highet etaient normales quoiqu'on ait releve la presence occasionnelle d'ectoparasites. Toutes les ombres prises dans Ie ruisseau Mud (un tributaire d'eau claire du ruisseau Minto)et maintenues dans des cages dans Ie ruisseau Minto pendant la m~me periode de 5 jours en septembreont survecu tandis que 16 % des poissons (5 sur 32)du ruisseau Mud maintenus pendant ce temps dans Ie ruisseau Highet sont marts en mains de 96 h.On attribue cette mortalite 8 une charge intolerable de stress decoulant des effets combines de la capture, du transport,de la mise en captivite et de l'exposition 8 des sediments en suspension et a des variations de temperature dans Ie ruisseau Highet. [ [ [ [ [ [ [ [ D t l".. C [ [ [ [ [ ~L [ D [' [ [ [ C [ o ~D ~ D C [ c [ l [~ [. [ -XVll - Quoique les valeurs d'hematocrite quantifiees chez les poissons en cage aux deux sites etaient semblables,les valeurs moyennes du glucose dans Ie plasma chez les poissons maintenus pendant 4 jours dans Ie ruisseau Highet en aoQt etaient de 30 %superieures a celles des specimens gardes dans Ie ruisseau Minto au meme moment.En septembre,des ombres capturees dans les ruisseaux Minto et Mud et mises en cage dans Ie ruisseau Highet ont subi une augmentation de 100 %des niveaux moyens de glucose dans Ie plasma par rapport aux valeurs du groupe correspondant garde dans Ie ruisseau Minto.On croit que ces differences tiennent aux conditions aquatiques plus stressantes (charges de sediments en suspension et differences de temperature plus prononcees)dans Ie ruisseau Highet par rapport au ruisseau Minto. On conclut que l'exposition a court terme de l'omble arctique a des concentrations subletales de sediments organiques ou inorganiques en suspension peut causer un certain nombre d'effets y compris des reactions de stress aigu.Tenant compte de ces decouvertes,il est necessaire de mieux comprendre l'incidence environnmentale des sediments provenant d'exploitation de gisements alluvionnaires sur les capacites d'adaptation immediate et a long terme (y compris l'alimentation et les autres reactions de comportement,la resistance aux maladies,la croissance et Ie bien-etre chronique)de cette espece de poisson sensible. [ [' [ [ [ c c c [ [ [ [ [ INTRODUCTION Placer mining activity in Yukon Territory has increased appreciably during the past few years.The exact impact of current and past mining operations on the fisheries resources in the Pacific and Yukon Region is not clearly understood; however,recent studies have shown site-specific evidence of damage to aquatic life and habitat (Mathers et al.1981;Singleton et al.1981;Weagle 1982). Adverse effects attributed to suspended and deposited sediment loads in receiving waters downstream of placer mining activities include degraded water quality (Knapp 1975;Anon.1981;Mathers et a1.1981);reduced numbers of benthic invertebrates (Anon.1979a;Mathers et al.1981);habitat disruptions and reduced numbers of Arctic grayling (Thymallus arcticus)and other fish species (Knapp in prep.;Anon.1979;Weir 1979;Mathers et al.1981;Singleton et al.1981; Weagle 1982).Despite.this evidence of threat to Yukon fisheries,present data concerning direc t evidence for the deleterious effects of suspended sediment on grayling and other sensitive aquatic species native to these waters are insufficient to permit a clear understanding of the impact of placer mining sediments on Arctic grayling. Earlier studies (Herbert and Merkens 1961;Anon.1965;Neumann et al.1975; O'Connor et al.1977;Noggle 1978)have reported that sediment suspended in water can cause acute lethal or sublethal effects toward fish.Although some non-salmonid fish species have been shown to survive short-term exposures to suspended sediment strengths as high as 100,000 mg·L-1 (Wallen 1951),bioassays conducted by Noggle (978)indicated that salmonid fish tolerance to natural stream sediment varied seasonally and that suspended sediment concentrations as low as 1,200 mg~L-1 could be acutely lethal to underyearling salmonid fish. Additionally,Noggle's (1978)findings demonstrated that lower sediment strengths could be stressful to these fish. The present studies were undertaken to provide an understanding of the acute lethal tolerance of Arctic grayling to placer m~n~ng sediment under both laboratory and field conditions;and to determine if short-term exposures to sublethal sediment strengths caused certain deleterious effects (gill histopathologies,impaired respiratory capacity,reduced tolerance to temperature extremes,stress responses)to these fish.The influence on these responses of differing sediment types (inorganic "paydirt"fines and organic "overburden"soil) found suspended in stream water as a result of placer mining activities (Anon. 1981;Emond 1982),and of seasonal changes in photoperiod and water temperature to which Arctic grayling were acclimated,were also examined in laboratory tests. The acute lethal and sublethal responses of laboratory-reared grayling to the reference toxicant pentachlorophenol (Davis and Hoos 1975)were determined in concurrent bioassays in order to relate the nature and extent of effects to those ascertained for other salmonid fish species with this contaminant. The acute bioassay tests to which these gray1 ing were subjected were based upon procedures developed previously for evaluating the short-term impact toward salmonid fish of a variety of aquatic contaminants or other environmental stressors (McLeay and Gordon 1980;Wedemeyer and McLeay 1980.As part of this [ - 2 - [ [ investigation,short-term in-situ bioassays were conducted on each of two occasions (August and September 1982)with wild underyearling grayling held captive in a Yukon clearwater stream (Minto Creek),and in a tributary stream (Highet Creek)downstream of active placer m1n1ng..It was hoped that these laboratory and field studies would provide a better understanding concerning the direct effects of placer mining sediments on the acute tolerance and short-term adaptive capabilities of juvenile Arctic grayling. [ [ [ MATERIALS AND METHODS LABORATORY STUDIES [ Fish collection D [ c [ [ young-of-the-year Arctic waters within the Yukon These fish,captured by fry)to 5.0 cm (l-g young Upon capture,fish were placed in plastic "laundry"baskets lined with fibreglass mesh screen,and held in the stream from which they were seined until sufficient numbers (800-1,000)were collected for shipment.Groups of 50-100 individuals were placed in creekwater within 20-L plastic bags,and provided with an oxygen atmosphere (cylinder 02).These water bags were placed in coolers and packed with ice at the earliest opportunity.Fish were trucked to Whitehorse (Yukon Territory)and air-expressed to Vancouver for the controlled laboratory bioassays.A total of five separate shipments were made. A heterogeneous population of approximately 5,000 grayling were collected from northern British Columbia River drainage basin during July and August,1982. seining,ranged in size from 1.5 cm (0.03-g swimup fingerlings),depending on collection site and time. Fish rearing Upon receipt at the Vancouver laboratory (B.C.Research),fish were transferred to an outside fibreglass hatchery trough (swimup fry)or to four outside 1000-L semicircular fibreglass tanks (fingerlings).Water supply to these tanks was Vancouver City dechlorinated tap water,heated and regulated to a constant temperature of 150 +1 0 C.The minimum water exchange rate to each tank was 2 L"g-l fish per day throughout the duration of this study.Additionally, fish-loading density in each tank was held below 2.5 g"L-1 to ensure that grayling were not overcrowded (Sprague 1973). Initially,fish were fed Biodiet No.1 (0.6 mm crumble size;Bioproducts Inc.,Warrenton,Ore.)supplemented with live brine shrimp.Food was offered 8-10 times daily,and trough/tanks siphoned daily to remove excess food and faeces. Due to difficulties encountered in encouraging the younger (swimup fry)grayling [ [ [ [ .[ [ [ [' [ [ [ [ c [ [ L [- [ - 3 - to feed,the trough-reared fish were also offered finely-ground beef heart, freeze-dried pulverized tubifex worms,live daphnia (Daphnia pulex),Oregon Moist mash and canned salmon. Feed crumble size was increased to 0.6-0.8 mm (Biodiet NO.2)during late August.This ration was gradually replaced with Oregon Moist pellets (OMP;1.6 mm)supplemented with twice-weekly feeds of live brine shrimp. Fish were size-sorted and transferred to clean tanks at 4 to 6 week intervals.Water temperature to which these fish were acclimated was maintained at 150 +1 0 C until December 1 (i.e.until all bioassays with IS oC-acclimated fish were completed).At this time,the water temperature within each of three outdoor tanks holding the remaining stock of grayling was decreased gradually (2 0 c day-I) using increasing flow rates of untempered (50 +O.SOC)Vancouver City dechlorinated tap water,until this colder temperature was attained.Grayling were acclimated to this water temperature for a 7-week period prior to the final series of bioassay tests.Throughout this period,fish were fed twice daily an excess ration of OMP together with freshly thawed sockeye salmon (Oncorhynchus nerka)eggs. Sediment collection A 200-kg sample of inorganic sediment was collected from a Highet Creek placer mine site on August 10,1982.This sample was coarse-screened on-site from a seam of near-bedrock material being actively sluiced,and particle sizes <2 mm retained.The nature of this sediment was characteristic of that commonly referred to by placer miners as "paydirt"(Emond 1982).The sample was transported to Vancouver in new 20-L sealed plastic buckets,whereupon it was mixed thoroughly (240-L plastic barrel),returned to the buckets and stored at 4 0 C until required for bioassay tests. A sample of organically-rich overburden material weighing approximately 200 kg was obtained during August from a site alongside Minto Creek where the vegetation had recently been stripped away.This dark-brown "muck"(80%moisture content)was also transported to the Vancouver laboratory in new (sealed)20-L plastic pails.The sample was mixed in a 240-L plastic barrel,returned to pails and stored in the dark at 4°C until required for testing. Sediment preparation and analyses Preliminary examination of the inorganic paydirt material indicated that the majority (>98%)of this sample was comprised of particles >1.0 mm (i.e.too coarse for the present study).Accordingly,a procedure was derived which reduced the sample to sediment fines.Quantities of paydirt required for each bioassay test were oven-dried (SOoC)to constant weight.Measured amounts (200 ml =280 g)were then pulverized for exactly 2 min using a vibratory ring pulverizer (TMS r - 4 -[ (' Engineering,Vancouver).A1iquots of pulverized paydirt were canbined and stored L. in polyethylene bags until used. Portions of each of these two prepared sediment types were analysed for the following characteristics:particle size distribution;particle shape;moisture content;volatile and fixed residue;rate of oxygen uptake;and major and trace inorganic components. Preliminary tests with the organic muck indicated that wet screening of this material to select particle fines was impractical.Quantities required for each bioassay test coarse-screened only to remove rootlets and woody debris from This pre-sorted undried organic overburden material was held in beakers until used for the bioassay tests. sieving or fine diHicu1 t and were therefore the humic soil. covered plastic [ [ [ Two 280-g samples of paydirt (sub-samples of portions prepared for the 4-day survival tests and stress bioassays)were examined for particle size distribution.Each sample was wet-sieved,oven-dried (SOoC)and mechanically agitated for 10 min through a standard series of Tyler sieves.Percentage weight of paydirt retained on each sieve was calculated (Anon.1972). A 300-g portion of coarse-screened organic muck was wet-sieved in SO-g increments.The oversized (+400 mesh)material retained was then oven-dried (SOO C)and rolled out with a stainless steel rolling pin (to break conglomerates). The resulting material was mechanically agitated (10 min)through sieves,and calculations made of the percentage weight retained on each. The appearance of each sediment type was examined microscopically.Both dry and wet (suspensions in water)preparations were viewed under dissecting (SOX)and compound (400X)microscopes. Moisture content of each test material was determined by drying SOO-g portions at lOS o c to constant weight.Their volatile and fixed components were ascertained by igniting each sample at SSOoC to constant weight (Anon.1980a). The oxygen uptake rate at lS o C for each sediment type was measured according to a procedure used previously (Anon.1979b)for evaluating dredged sediments. Fixed volumes (30 ml)of material were added to SOO-m1 Erlenmeyer flasks containing SOO m1 of oxygen-saturated freshwater (Vancouver City dechlorinated tap water)at lS o C.Each flask was stoppered,shaken and allowed to remain undisturbed for 24 h at this temperature.Initial and final dissolved oxygen values for the overlying water were measured (Delta Scientific Model 1010 portable oxygen analyser with mechanical agitator)and oxygen uptake rates calculated. The major and trace inorganic constituents of each test .material were determined by plasma spec trographic analysis.Dried (lOSOC)preparations were digested using a combination of acids (HF,HC1,HN03'HC104)and the resul ting solutions analysed for metals using an inductively coupled argon plasma spectrograph (Can Test Ltd.,Vancouver,B.C.). [ [ [ [ [ [ [ I -[ [ [ [' [ [ [ [ c c c [ c [ [ [ - 5 - The_interrelationship of nonfiltrable residue (suspended solids),total residue and turbidity values for suspensions of each sediment type in freshwater was examined.A range of concentrations (nominally 0-50,000 mg sediment:L-1,dry weight basis)of paydirt or overburden material in freshwater (Vancouver City dechlorinated tap water)was prepared using separate 1-L plastic bottles. Aliquots (l00-m1 volume)were taken from each bottle for determinations of total nonfi1trable residue,total residue,and turbidity (formazin turbidity units - FTU).Each aliquot was taken immediately after vigorous agitation of the sample bottle.Aliquots for turbidity analyses were re-agitated just prior to examination.All analyses were performed according to Standard Methods (Anon. 1980a). Recycle test tanks Thirty 50-L capacity recycle test tanks were constructed for use in the acute survival bioassays,temperature tolerance tests and stress bioassays.The basic design for each tank was according to Noggle (1978).The body of each tank,made of 6-mm translucent plexiglass sheeting (transparent sheets sand-blasted to reduce visual disturbances to fish in clear solutions),measured 41 X 37 X 36 cm.A steeply sloping conical-shaped bottom ensured that all settleable solids would be collected and re-circulated.During operation,the test suspension in each tank was withdrawn continuously from this cone through a pump (Little Giant Model 1-42) at a rate of 10.3 +0.3 Lomin-l (mean +SD;n =20),and respilled onto the surface of the suspension (see Fig.1).- A rectangular fish basket,made of soft-mesh nylon netting framed with stainless steel rods,was constructed to fit the body of each tank.These baskets were used to contain fish and to raise them for periodic observations or for sampling. Bioassays using these tanks were conducted in a temperature-controlled room removed from general laboratory disturbances.Overhead incandescent lighting, regulated by photocell,provided a natural photoperiod for all tests.Lights were brightened/dimmed gradually OO-min automated rheostat)at the start and end of each daily cycle to simulate natural conditions. Acute survival tests Fish acclimated to l5 0 C A study was conducted to determine fish mortalities and gill histopathologies associated with acute (up to 4-day)exposure to suspensions of paydirt or overburden sediment fines.Five underyearling grayling acclimated to laboratory water at 15 0 C for 7 weeks were placed randomly in each of a series of 50-L volumes of these suspensions within the recycle test tanks.Nominal strengths (dry weight basis)of paydirt to which these groups were exposed ranged from 50 to 250,000 mg sedimentoL-l,and from 50 to 50,000 mgoL-1 for the overburden suspensions.Each suspension was prepared by mixing a pre-weighed amount of test material into the tank while the freshwater was re-circulated.Vancouver City dechlorinated tap water (at l5 0 C)was used for preparing all suspensions and as the control water - 6 - (no sediment added).Overhead airconditioning was regulated to hold the temperature of each suspension within values to which these fish were acclimated. Initial tests with the suspensions of overburden indicated that nominal strengths 10,000 or higher (dry weight basis)fouled the pumps.Consequently, agitation of these higher organic sediment strengths was maintained by upwelling compressed air through the apex of the conical bottom of each of these test vessels. Water temperature,pH,dissolved oxygen content (mg O2 ~L -1)and conductance (umho·cm-1 )values for each suspension were monitored daily throughout a 96-h test period,together with fish survival and behavioural observations (surfacing, coughing,swimming activity).Upon completion of this period of exposure, surviving fish in each test suspension were netted sequentially and their fork length (cm)and wet weight (g)determined.The caudal peduncle of each fish was severed,and blood collected in heparinized microhematocrit glass capillary tubes.All blood samples from each group of fish were collected within 5 min. Blood samples were centrifuged (12,500 rpm;3 min)and hematocrit values (Fig.2) measured.Plasma portions were separated and stored frozen (-20 0 C)until analysed (10 uL aliquots)for glucose content (Beckman Glucose Analyser 2). Gill tissue was dissected from each fish and placed immediately in Bouin's fixative.These tissues were transferred 24 h thereafter to 95%ethyl alcohol. Subsequently,selec t groups of these tissues (gills from three fish held in 01100,1,000,10,000,and 100,000 mg·L-1 paydirt or 0,100,5,000 and 50,000 mg·L- overburden)were paraffin-embedded,sectioned (6 um)and stained (hematoxylin/ eosin)for histopathological examination. A 100-ml aliquot of each test suspension was taken from the end of the pump outlet tube (paydirt suspension)or from the centre of the tank (overburden suspensions)at the termination of the 4-day fish survival tests.These aliquots were dried and analysed for total residue content (Anon.1980a).Results were expressed as final suspended residue concentration (mg sediment·L-1). Upon completion of the 4-day exposure tests .with grayling and paydirt suspensions,ten hatchery-reared rainbow trout (Salmo gairdneri)swimup fry (0.5 + 0.1 g;3.4 +0.3 cm)were added to each test suspension within the recycle test tanks.These fish were acclimated to Vancouver City dechlorinated tap water since their receipt as eyed eggs.The survival of these salmonid fish was monitored daily throughout a subsequent 4-day period of exposure. Exposure of one group of Arc tic grayling to a high strength of suspended paydirt fines (50,000 mg·L-1)was continued for a total of 16 days,during which time daily observations of fish were made.This suspension was recycled continuously throughout the 16-day test period,and water temperature was held at 150 +1 0 C. A final group of five grayl ing accl imated to 15 0 C was examined for 4-day survival in a high (100,000 mg·L-1)concentration of suspended paydirt prepared by [ [ [ [ [ - C [ [ [ [ [ L I ~.[ [ [ [' [ [ [ c c -,c [ [ L [. [ - 7 - sieving only (no pre-grinding).Sediment used for this test was that portion of dried material which passed through an O.S rom pore sieve.A sample of this sieved test material was analysed for partic Ie size distribution. Fish acclimated to SoC The acute tolerance of SOC-acclimated Arctic grayling to suspended paydirt fines was examined under controlled laboratory conditions.Groups of ten fish held previously at SO +10c for seven weeks were transferred from the outside holding tanks to separate recycle test tanks containing SO L of inorganic suspensions ranging in concentration from SOO to 100,000 mg"L-l.Fish survival, water temperature,pH,conductance and dissolved oxygen content in each tank were monitored daily throughout a 4-day period of exposure.Overhead airconditioning was adjusted to maintain the water temperature in each test tank within the range to which these fish were acclimated (SO +O.SOC).Other conditions and procedures were according to those described previously. Following a 96-h exposure,individual fish surv1v1ng in each test suspension were netted rapidly (within a 7-min period).Lengths and weights were recorded, and blood samples collected and processed (as described previously)for hematocrit,leucocrit (see Fig.2)and plasma glucose determinations.' The consistency with which differing strengths of inorganic sediment remained suspended within the recycle test tanks was examined during this 4-day test. Aliquots (100 ml)of each suspension were withdrawn from the centre of each tank for total residue analyses at each of the following times after their introduction:0,O.S,S,24,48,72 and 96 h.Additional aliquots were taken from each tank at 48 h in order to assess the dispersal pattern for each recirculating suspension.These samples were taken from each tank at the following locations:inflowing suspension (end of pump outlet tube);surface (centre of tank);mid-depth (centre of tank);and near a bottom corner of the net enclosure.Each aliquot was analysed for total residue concentration (Anon. 1980a)• Temperature tolerance tests Fish acclimated to lS o C The effect of suspended.paydirt or overburden material on the critical thermal maxima (upper lethal temperature tolerance)for underyearling Arctic grayling acclimated to lS oC (for 9 weeks)was determined in separate studies. Basic test procedures for this bioassay were according to those described previously (McLeay and Howard 1977;McLeay and Gordon 1980). Ten grayling were transferred randooly to each SO-L test suspension within each of a series of recycle test tanks.Test apparatus and procedures for preparing each suspension were identical to those given for the acute survival tests.Nominal concentrations of inorganic paydirt to which fish were exposed ranged from 2S to 100,000 mg"L-1 ,and from SO to SO,OOO mg"L-1 for the organic overburden. - 8 - The temperature of each test suspension was initially lS o C.This temperature was increased progressively at a controlled rate of 1 0 coh-1 (electric baseboard heaters coupled with a thermostatically-controlled immersion heater in each tank) until all fish in each tank were dead.The temperature of each test suspension was recorded (+O.lOC)at the time of death of each fish.These fish were removed and measured (length,weight).Aliquots of each suspension were then taken from the centre of each test vessel for analyses of final suspended residue content. Fish acclimated to SoC The effect of paydirt suspensions on the critical thermal maxima for grayling acclimated to SoC for 9 weeks was examined.Groups of ten fish were transferred from a rearing tank to recycle test tanks containing nominal paydirt suspensions ranging from 100 to 50,000 mg °L -1.,The temperature of each test suspens ion was initially SoC,and was increased at 10 Coh-1 until all fish were dead.Other test procedures and conditions were identical)to those used for the temperature tolerance tests conducted with lS o C-acclimated grayling. Sealed jarbioassays Fish acclimated to lS oC Sealed jar (residual oxygen)bioassays were conducted with juvenile grayling acclimated to lS o C for 12 weeks.Basic test procedures were those developed for use with kraft pulpmill effluents (McLeay 1976;Gordon and McLeay 1977)and applied subsequently with other aquatic contaminants (McLeay and Gordon 1980). [ r [ [ [ [ [ [ c c Each jar was inverted at 20-to 30-min intervals throughout the test period in order to re-expose fish to any settleable solids.Control jars (freshwater only)were treated accordingly.The survival or death of fish was determined on these (and more frequent)occasions.Upon the death of each fish,water temperature and time to death were recorded.The residual oxygen level in each suspension was measured using a portable oxygen meter (Delta Scientific Model No. 1010)with mechanical agitator. For each concentration of paydirt or overburden examined,ten replicate jars were prepared (identical weights of sediment added to each).Two replicate sets of ten control solutions (freshwater only)were included with each series of sealed jar tests conducted with paydirt or overburden sediments.Air-saturated freshwater (Vancouver City dechlorinated drinking water)at 20 0 C was added to each jar and the fish introduced.Each jar was then filled completely with water,and sealed (plastic lid)to exclude air. Grayling weighing approximately 10 g were selected for these bioassays. sealed jar tests were conducted at 20 0 C using 1.9-L glass jars,one fish per (fish-loading density,S goL-l)(McLeay 1976). The jar [ [ [ [ [ The initial (maximum)and final (minimum)suspended residue concentrations to which fish were exposed during these tests were determined.-At the time of the [ [ c [ [ c c D c [ l [~ [ - 9 - bioassays,one additional jar conta1n1ng each test suspension was prepared,and a 10-g fish added.Upon the inversion of each jar,an aliquot of each suspension was extracted (by syringe)from the jar's centre.This procedure was repeated after the jar was left undisturbed for 30 min.Each aliquot was analysed for total residue concentration. Fish acclimated to SoC Underyearling grayling acclimated to SoC freshwater for 11 weeks and weighing approximately 10 g were selected for this study.Sealed jar bioassays with differing strengths (100-100,000 mg-L-l)of suspended paydirt fines were conducted at 10o C,using air-saturated freshwater adjusted to this temperature overnight as the control or test (dilution)water to which fish were exposed.Otherwise,test apparatus and procedures used for this bioassay were identical to those employed in the previous sealed jar test with grayling and paydirt. Acute stress bioassays Controlled bioassays were performed to determine the concentrations of paydirt and overburden suspensions which are acutely stressful to Arctic grayling.Basic test procedures were those proven effective for determining threshold strengths of a variety of aquatic contaminants which cause stress responses (elevated blood sugar levels,decreased numbers of circulating leucocytes)with other salmonid fish species (McLeay 1977;McLeay and Gordon 1977, 1979,1980). Groups of ten underyearling grayling acclimated to lS o C for 12 (overburden bioassays)or 13 weeks (paydirt bioassays)were transferred from the outside tanks to a series of indoor recycle tanks containing freshwater (at lS 0 C)only.Fish were left undisturbed in these tanks for a 48-h period in order to adapt to the stress caused by this transfer.Thereafter,weighed portions of paydirt or overburden sediment were added to each tank at 20-min intervals.Tanks for each treatment were chosen randomly.For each test (paydirt or overburden material), two tanks were selected as controls.Nominal concentrations of paydirt to which these fish were exposed ranged from SO to 100,000 mg'L-1,and from SO to 20,000 mg-L-l for fish held in suspensions of overburden material. Each group of ten grayling was sacrificed for blood sugar and leucocrit determinations after a 24-h exposure to each sediment suspension.The control groups were sampled just prior to and again just subsequent to the sampling of all experimental g'roups to ensure that no changes in the stress responses measured were caused by sampling disturbance.Sampling procedures and methods for determining plasma glucose,hematocrit and leucocrit values for each fish were identical to those described previously in this report. -10 - Reference toxicant tests The response of the laboratory-reared Arctic grayling to the reference toxicant pentachlorophenol (Davis and Hoos 1975)was determined at the time that these bioassay tests were conducted.Fresh stock solutions of pentachlorophenol were prepared by dissolving 100 mg dry powder (Aldrich Chem.Co.Inc.;Lot No. 122047;purity >99%)in 10 ml of 2%NaOH,and diluting to 1 L with deionized water (Alderdice 1963).These concentrated stock solutions were diluted ten-fold as required for each bioassay. The acute lethal tolerance to pentachlorophenol of grayling acclimated to ISO or SoC was determined just prior to the start of the acute survival tests with paydirt sediment.Groups of ten fish were transferred from stock tanks to 4S-L glass aquaria containing pentachlorophenol concentrations (diluted with Vancouver City dechlorinated tap water)ranging from 30 to 120 ugoL-l.Test temperature tcii:· these static bioassays was held at that to which the grayling were acclimated (ISO or SOC).Fish survival in each test solution was monitored daily throughout a 4-day test period. The effect of sublethal and lethal concentrations of pentachlorophenol on the upper lethal temperature tolerance of juvenile grayling acclimated to lSoC was ascertained.Groups of ten fish were trans ferred from a stock tank to recycle test tanks containing pentachlorophenol strengths of 0 (freshwater control),2S, SO and 80 ugoL-l freshwater.The temperature of each test solution was increased from an initial value of lSoc at a rate of 10coh-l until all fish died,and the temperature at time of death of each fish determined.Conditions and procedures for conducting this bioassay were identical to those described for the temperature tolerance tests performed with lSoC-acclimated grayling and sediment suspensions. The effect of pentachlorophenol on the tolerance to hypoxia of lSoC- acclimated grayling was determined by sealed jar bioassay.Materials and methods were those described earlier.Residual oxygen levels at death were determined for fish held in jars containing pentachlorophenol strengths of 0 (two freshwater control groups),3S,SO and 80 ugoL-l • The effect of sublethal concentrations of pentachlorophenol on acute stress responses for lSoC-acclimated grayling was examined at the time and according to procedures described for the stress bioassays carried out with grayling and sediment suspensions.For these tests,four groups of ten fish were exposed to the following strengths of this reference toxicant for 24 h:0 (freshwater control),20,3S and SO ugoL-l.Plasma glucose,hematocrit and leucocrit values for each of these fish were measured as described previously. The length (cm)and wet weight (g)of each fish exposed to pentachlorophenol were determined at time of death or upon termination of each bioassay test. Statistical analyses The condition factor (K)of juvenile grayling used in each bioassay test was determined as follows:K =cW·L-3 where c is a constant (l00),W is weight (g)and L represents fork length (cm)(Carlander 1969)0 [ f[ [ [ [ c [ o c [ [ [ [ [ L ~[ [ [ [ [ -11 - Mean and standard deviation (SD)values for fish length,weight and condition factor were calculated for each bioassay.Mean +SD plasma glucose,hematocrit and leucocrit values determined for each group of fish receiving identical treatment were also determined.Additionally,mean (+SD)temperatures at death (temperature tolerance and sealed jar tests),times to death and residual 02 values at death (sealed jar test)were calculated for each control and test group.For values shown graphically,the 95%confidence interval of each mean was determined. The median effective concentration (EC50 value)of each sediment type causing a net significant response for 50%of the fish treated identically in each bioassay (temperature tolerance test,sealed jar and acute stress bioassays)was calculated according to established procedures (Sprague 1968;McLeay and Howard 1977;McLeay and Gordon 1980).Relevant values determined for each test fish (i.e.temperature at death,temperature tolerance test;time to death and residual 02 at death,sealed jar bioassay;plasma glucose and leucocrit,acute stress test) were examined to determine the number of responses for each treatment outside of the 95%confidence interval for the corresponding group(s)of control fish. Depending on the suitability of the data derived in this manner the EC50 value for each test was calculated,together with its 95%confidence interval (Stephan 1977). [ c ... [ The acute median lethal concentration toxicant pentachlorophenol,as determined acclimated to 50 or 150 C,W'as calculated interval)using the computerized LC50 program (96-h LC50 value)for the reference with groups of juvenile grayling (together with its 95%confidence of Stephan (1977). [ I' L l [- l FIELD STUDIES Study area General The Highet Creek and Minto Creek sites chosen for the in-situ caged fish bioassays were selected following an aerial and ground reconnaissan;e-of some nine creeks in Central Yukon on June 26 and 27,1982.The study area (Fig.3)lies in the Selwyn Basin portion of the Canadian Cordillera,and is underlain by sedimentary limestone skarn and quartzite schist rocks of the Windermere Group (Tipper et al.1982).Quartz is the main mineral in this mineralogically complex rock,although diorite and granitic rock protrudes through the older schists in several areas. The surficial deposits in the area can be divided into three recognizable types:the upper,post-glacial unit consisting of recent and terrace gravels;the glacial unit consisting of till or glacio-lacustrine silts and sands;and the lowermost pre-glacial unit making up the deep,terrace and high level gravels. Several of these deposits,in particular the deep gravels,glacio-lacustrine silts and sands and the till,are exposed in Highet Creek (Cairnes 1915). -12 - The pre-glacial deep and terrace gravels are the most productive in terms of placer gold.They cover the hummocky bedrock of the valley bottom and are from 3 to 8 metres thick.The lower 2-4 m of the deep gravels are commonly stained with manganese oxide and iron oxide.The black to red oxides occur in the gravel matrix and as a stain on the clast surfaces.Some manganese oxides occur in crystals (Emond 1982). Glacio-lacustrine silts and sand gravels varying in thickness from 3 to 25 m overlie the pre-glacial gravels.These sediments are usually finely laminated dark grey sil t layers interlaid with brown sand.The silts were deposited in a shallow lake that was formed when ice,moving westward up Minto Creek,protruded into the lower part of Highet Creek (Bostock 1939). The glacial till of Highet Creek varies in thickness from 1 to 4 metres.The gravel contains pebbles of assorted rock types.The matrix is a yellowish brown, silty or gritty clay (Emond 1982).Recent gravels approximately 2 metres thick are found in the lower part of the creek valley.The gravel contains well-rounded pebbles and cobbles that are less than 7 cm in diameter.Trace geochemical analyses indicate that gold,tungsten,chromium,iron,manganese,titanium,zinc and zirconium are the most abundant heavy elements in the Highet Creek gravels. Tin,arsenic,cadmium,and mercury were not detected (Emond 1982). In the Mayo-McQuesten area,gold was first found on sand bars of the Stewart River in 1883.Prospecting of creeks draining the upland led to the discovery of gold,in 1901,in Duncan Creek.Gold was first discovered on Highet Creek in 1900 by Warren Hiatt.Mining began in 1903 on bench claims on the right limit of the upper part of the creek.Several operators mined gravels in the creek bottom during the period 1916-1946.During this early period,a dredge was worked unsuccessfully on the creek for one season.Since 1960,mining on Highet Creek has consisted of three small individual operations which utilize earth-moving equipment and large sluice boxes.Highet Creek has been one of the leading gold producing creeks in the Mayo-McQuesten area. Highet Creek Highet Creek originates on the upland (maximum elevation 1825 m)between the McQuesten and Stewart rivers,and flows to the southwest through a narrow valley into Minto Creek (Fig.3).Although ungauged,this high gradient stream (elevation change 90 m-km-l )likely exhibits a seasonal hydrograph similar to other small creeks subject to placer mining in the Mayo area.In undisturbed creeks,peak flows occur during May and early June in response to snowmelt (Anon. 1980b).However,during the summer months the flows in Highet Creek are regulated,and hence may vary considerably from day to day in response to placer mining activity.Winter freeze-up of this creek usually begins in November and extends to April (Allen and Cudbird 1971). [ -[ [ [ [ [ .... [ [ [ [ [ [ [ During the open water period,Highet Creek is frequently turbid mining and erosion of previously mined sections of the valley. suspended sediment load carried by the creek from June to October due to placer Most of the is created by -[ [ [ [. [ c [" [ [ D 0 C C C C C [ [ [~ C~ l -13 - mining operations 1n the upper third of the valley.In the lower sections of the creek,the suspended sediment load consists of that portion of the mine effluent remaining after passage through several settling ponds located 3.0 km upstream of Minto Creek (Fig.3). No information on the former use of Highet Creek by fish is available.A preliminary species abundance/habitat study conducted during the summer of 1982 found only limited numbers of underyearling Arctic grayling within Highet Creek, whereas Minto Creek and other Minto Creek tributaries (Bennett Creek,Mud Creek) contained appreciably larger populations of Arctic grayling and other species of fish (Birtwell et al.1983). The site in Highet Creek selected for in-situ caged fish studies was located 0.25 km upstream of the junction with Minto Creek (Fig.3).Biophysical characteristics determined for this site during these field bioassays are provided in Appendix 1. Minto Creek Minto Creek flows eastward into the Mayo River through an upland part of the Yukon Plateau that lies between the Stewart and McQuesten river valleys (Fig.3). Minto Creek originates at Minto Lake (elevation 685 m)and follows a winding l6-km course into Wareham Lake (elevation 580 m).The stream gradient is generally low (l.5 m'km-l ),particularly in the upper third where it is further reduced by a series of beaver dams.Although ungauged,Minto Creek likely exhibits a seasonal hydrO-graph similar to other lake-fed streams in the Mayo area.Peak flows occur shortly after breakup in late Mayor early June whereas low flow usually occurs in February or March (Anon.1980b).The valley of Minto Creek is undisturbed with the 'exception of a site approximately 1.5 km below Minto Lake that was placer mined briefly in 1980. During the summer months,water in Minto Creek above its confluence with Highet Creek is clear to slightly turbid.Below Highet Creek,Minto Creek water is frequently turbid from June to October each year due to the placer mining activities on Highet Creek. The control site in Minto Creek selected for the in--situ caged fish studies was 0.5 km ups team of the junction with Highet Creek-,-ata location that was 'similar in stream flow and other characteristics,except suspended solids,to the test site in Highet Creek (Appendix 1).This site was chosen for its clear water and its proximity to the Highet Creek site. Fish collection For the in-situ caged fish studies,several hundred wild underyearling Arctic grayl ing wer;captured from Minto Creek for August bioassays,or from both Mud Creek (Fig.3)and Minto Creek for September bioassays.Those fish taken from Minto Creek were collected 0.5-0.8 km upstream of the junction with Highet Creek. -14 - Fish were captured from shallow pools and riffles,using one or more seine nets.Upon netting,fish were placed in plastic holding pens lined with fibreglass screening (allowing free flow of creekwater).All captured fish of suitable size were held for 1 to 2 days in Minto Creek (just upstream of the control site)prior to their transfer to cages. In-situ bioassays Test apparatus Ten net enclosures were used for the in-!ltu bioassays.Each enclosure (30 cm deep by 45 cm diameter)consisted of two aluminum rings covered with soft nylon mesh (4 mm).A drawstring in the mesh at the top of the enclosure could be opened to inspect the fish (Fig.4). At each site (Fig.5 and 6),five enclosures were located adjacent to one another at the upstream end of a pool.Each 50-L enclosure was suspended in the water column,5 to 10 cm below the surface,between three tubular iron posts (Fig.4).The position of the posts provided stability and allowed each enclosure to be lifted independently for inspection,while at the same time ensuring free circulation of water. Water quality Water samples were collected at both the test site (Highet Creek)and the control site (Minto Creek)during the August and September caged fish tests. Samples (300-400 ml)were collected hourly at each site for the duration of the tests,using an ISCO automatic pump sampler.The sampler's intake port was located wi thin an empty fish cage submerged at each site.Samples were removed every 24 h and stored in plastic bot tles within coolers for shipment to the Environmental Protection Servic'e/Fisheries and Oceans laboratory at West Vancouver.The following characteristics were measured for alternate samples collected from each site,using procedures established by Environment Canada and Fisheries and Oceans (Anon.1979c):total residue;total fixed residue; nonfiltrable residue;total volatile residue;and turbidity (FTU). The particle size of nonfiltrable residue within Highet Creek was estimated from composite creekwater samples taken during both the August and September test periods.These samples were made by combining one hourly water sample selected at random from each of the test days.Each composite sample was analysed by Soil Analysis Inc.(Vancouver)for particle size distribution,using the pipet method (Anon.1975). On seven occasions during the August and September test per iods,duplicate grab samples were taken from Highet Creek adjacent to the test enclosures.These samples were collected to coincide with samples being drawn from within the enclosures by the automatic sampler.Analyses included turbidity (FTU), nonfiltrable residue and total residue. [ I [ [ [ [ [ fl..,.·.·...U [ [ c [ [ [ ~[ [ [' [ [ [ [ c c c [ [ [ -15 - Two or more times daily during each in-situ bioassay,the following variables were measured near the net enclosures at each site:water temperature (oC),pH,_ and dissolved oxygen (mg 02°L-l).Water temperature was measured using a mercury thermometer.A portable YSI Model 57 dissolved oxygen·meter and a Corning Model 6l0A pH meter were used for the other field determinations. Several 150-or 250-ml water samples were collected in polyethylene bottles at the enclosure sites.Samples were collected at the beginning and end of each test period for alkalinity,hardness,Mn,Mg,Na,Ca,Si,and total metals (As,B, Ba,Cd,Cr,Cu,Hg,Ni,Pb,Sb,Sr,Zn,AI,and Fe)•Samples not requiring preservation were kept cool for return to the laboratory at the conclusion of each study period.All metals except Hg were preserved with 1 ml of HN03;Hg samples were preserved using a solution of H2S04 and K2Cr207'Analysis of all water samples was performed at the Environmental Protection Service/Fisheries and Oceans'West Vancouver Laboratory,using techniques specified (Anon.1979c). Experimental Short-term (4-to 5-day)in-situ bioassays were conducted at the Highet and Minto Creek sites on each of two ~sions -August 6 to 10,and September 10 to 15.These field bioassays were conducted twice to determine the effect,if any, of different water temperature regimes on the acute tolerance of grayling to suspended sediment. In orde r to ma intain a load ing dens i ty of approxima te ly 1 go L-1,groups of 14 to 26 fish were selected randomly from the holding pens on each occasion,and transported in plastic pails to each of four net enclosures at each site.The mean density of fish placed in each enclosure was approximately 1 gOL-I •During August,each cage received grayling captured from Minto Creek;whereas during the September tests,fish introduced to only two of the four cages at each site were captured from Minto Creek.The remaining two cages at each site on this occasion received Mud Creek fish only. Fish within each cage were observed hourly for the initial 4-h period,and twice daily (between 0900 and 1100 h,and between 1900 and 2200 h)thereafter until the tests were terminated.Each inspection was carried out by raising the cage until its bottom was just below the surface of the water.Inspections were conducted quickly to minimize stress to fish.Any dead fish observed were removed and examined (including measurements of length and weight).Times to death and frequency (%)of fish mortalities were recorded.Fish in cages were not fed on either occasion. Upon completion of a 96-h (August bioassays)or l20-h (September bioassays) period of exposure,all surviving fish in each cage were sacrificed.Fish were removed individually from each cage,and their fork length and wet weight determined.Gill arches (third or fourth)were removed from five fish selected randomly from each net enclosure,and mounted on glass slides in polyvinyl lactophenol.These tissues were later examined under dissecting and compound microscopes to detect any gross changes in gill morphology. -16 - For the September bioassays only,gill tissues of five grayl ing caged in Minto or Highet Creek for five days were removed and placed in Bouin's fixative. Gills from five untested grayling seined from Minto Creek were also taken and preserved upon fish capture.These 15 tissues were subsequently transferred to 70%ethyl alcohol,wax-embedded,sectioned (6 um)and stained (hematoxylin and eosin)according to standard practice.For each specimen,a number of pathomorphological changes involving the gill filaments and lamellae were rated on a scale of 0 (normal)to 4+(extreme pathology).Each specimen was coded and examined "blind"(without knowledge of treatment)to prevent biases from influencing the ratings assigned. Blood samples were taken from five fish selected randomly from each cage at the time of these autopsies.Each sample,collected in a heparinized microhematocrit tube,was centrifuged in the field (microhema tocrit centrifuge with 12-V battery and power converter),and hematocrit values determined.Plasma portions were separated and stored cool but unfrozen (held on ice)for subsequent glucose analyses.Other procedures for blood collection and analyses were according to those described previously. Samples of blood were taken from 19 and 30 underyearling grayling captured by seining from Mud Creek (August 9,1982)and Minto Creek (September 14,1982), respectively.These samples were taken within 5 min of fish capture.Blood hematocrit and glucose values were determined for each of these fish according to procedures used for caged fish. RESULTS AND DISCUSSION LABORATORY STUDIES Fish growth and condition All grayling air-expressed to Vancouver were alive and in apparently good condition upon their receipt.Efforts to initiate feeding of those fish received as swimup fry proved largely unsuccessful,resulting in a high rate of mortality. These fish mouthed the Biodiet ration offered,but spit it out.Offerings of other ration types (i.e.live Daphnia pulex or finely-ground beef heart)were also unaccepted by these very small fish.Unlike these findings,the larger fish (0.2-1.0 g)commenced active feeding within one week of their receipt.Biodiet ration no.1 was consumed vigorously by these fish and mortality rates were low. Cannibalism of smaller grayling was observed frequently,although size- sorting of surviving fish between the four outdoor rearing tanks appeard to mini- mize this problem.Fish transferred to these tanks fed actively on the commercial rations offered (Biodiet and,later,OMP).Live brine shrimp was readily taken by the fish.Growth of fish acclimated to the 15°C laboratory water supply (pH 6.8 * 0.2;conductance,15.7 ±1.2 umbo·cm-l ;nonfiltrable residue,<0.1 mg·L-l; residual chlorine,<0.01 mg·L-l;alkalinity,4.5 ±1.3 mg CaC03·L-l;EDTA hardness,5.0 ±0.3 mg CaC03·L-1)was rapid,with mean (*sD)weights of 2.4 ± 1.2 g by 6 weeks.The stock tank of larger grayling (sized-selected for the [ f '[ [ [ [ [ [ c ... C D c [ [ [ [ .[ [ The acute parentheses)to laboratory-reared [ c c [ [ c o c c c [ [ [ Co [ -17 - sealed jar bioassays)weighed 10.4 +1.3 g at 12 weeks.Mean condition factors determined for groups of lS o C-acclimated grayling sampled from the rearing tanks for bioassay tests were 0.8 to 1.0.These values are typieal of those determined for underyearling grayling collected from the Minto Creek drainage (Birtwell et ale 1983). Upon lowering of the water temperature to SoC,the rema1n1ng stock supply of grayling (divided between three outdoor tanks)ceased to feed.Offerings of OMp and brine shrimp were largely rejected during the subsequent S-week period. Thereafter,an attempt to promote feeding by introducing freshly thawed sockeye salmon eggs proved successful.Active feeding resumed at this time and continued until the bioassay tests with SOC-acclimated fish were terminated.Condition factors for grayling at the time of these bioassays were similar to previous values (0.8-0.9).The quality of the SoC water to which these fish were acclimated was as follows (based on weekly measurements):pH 6.7 +0.2; conductance,14.3 +2.2 umhoocm-1 ;nonfiltrable residue,<0.1 mg~L-1;residual chlorine,<0.01 mfoL-1;alkalinity,S.3 ~0.8 mg CaC0 3 °L-1;and EDTA hardness,6.2 ~1.2 mg CaC03°L-• No behavioural anomalies nor signs of disease were evident within the stock supply of grayling held at ISo or SoC for the laboratory tests.All fish deaths observed (other than those due to cannibalism)were attributed to starvation. lethal tolerance (96-h LCSO;9S%confidence interval in the reference toxicant pentachlorophenol determined for the grayling acclimated to ISo or SoC was as follows: lS o C fish:67 ugoL-1 (S7-77); SoC fish:61 ugoL-1 (48-71). All grayling acclimated to either temperature regime survived a 96-h exposure to pentachlorophenol strengths of 40 ugoL-l and lower.These LCSO values are within the range of tolerance for this respiratory inhibitor reported previously for populations of healthy hatchery-reared rainbow trout or coho salmon (Oncorhynchus kisutch)fingerlings acclimated to and tested in 10-120 c water with pH and hardness characteristics similar to that used in the present studies (Davis and Hoos 1975;McLeay and Gordon 1980).Results from these bioassays,considered together with the observations of the fish stocks,suggest that the condition and tolerance to aquatic contaminants of the laboratory-reared grayling were typical of healthy populations of young salmonid fish species at the time that each series of bioassay tests with suspended sediments was undertaken. Various workers have reported difficulties with the artificial propagation of grayling.Davis (1967)indicated that certain early investigators were only able to rear grayling fry to the fingerling stage when fish were supplied with creek water containing natural food.Others (Rawson 19S0)reported success with finely ground beef liver or heart,supplemented with goldfish food. As with the present study,other attempts to initiate feeding under laboratory conditions for Arctic grayling captured in the wild as swimup fry have proven largely unsuccessful (LaPerriere and Carlson 1973;Horler MS,1980).Part of the problems encountered by these investigators were likely due to the type -18 - (and crumble size)of commercial ration offered.Transfer stress associated with the capture and transport of swimup fry may also be implicated,together with any prior history of their feeding in the wild.Young grayling fry are thought to be planktivorous feeders',and their early feeding may be restricted by the relatively large size of food organisms available (Bishop 1971;Schmidt and 0'Brien 1982). Since the initial laboratory feeding of wild grayling captured as fry>0.2 g was achieved in the present studies using Biodiet,this commercial ration-appears to be adequate for this purpose.However,the use of other (non-commercial)food supplements (i.e.live brine shrimp for lSoC-acclimated fish;sockeye salmon eggs for SOC-acclimated fish)was also required to maintain feeding vigour. Characteristics of test sediments Particle size distributions for the paydirt and overburden sediments used in the laboratory bioassays are given in Table 1.Analytical results for each of the two preparations of paydirt fines were similar.Over 90%of the particles in each inorganic sample examined were <0.2 mm (i.e.fine sand,silt or clay);and approximately 70%of the test material was <O.OS mm (silt and clay).The majority (60-6S%)of the inorganic sediment sample to which grayling were exposed was comprised of particles less than 0.038 rom «38 um)in diameter (Table 1).Unlike this material,approximately SO%by weight of the organic overburden was made up of coarse material ()0.2 mm)and only 3%of this soil sample was very fine particles (<38 um).The remainder (47%)of the overburden contained particle sizes characteristic of fine sand or coarse silt.The particle size distribution of these test sediments is,in general terms,characteristic of overburden material found overlying paydirt gravel and of inorganic fines carried into downstream waters during sluicing operations (Anon.1981). Microscopic examination (400X)of the inorganic material indicated that the dry tan-brown sediment formed amorphous particles (electrostatic adhesion)which readily dissociated into minute particles upon addition to water.The dark-brown overburden sample was comprised of a considerable quantity of woody debris interspersed with soil particles of various shapes and sizes (very fine to coarse).Further detail concerning the shapes of these test materials could not be discerned by light microscopy,and scanning electron microscopy was not applied. Values for moisture content,volatile/fixed residue content and oxygen uptake rate (in freshwater at lS o C)for each test sediment are presented in Table 2. The moisture content for the overburden muck remained at 82-8S%throughout the period that this sample was stored.This sample was comprised of 96%volatile (organic)material;whereas the volatile component of the paydirt was only 4%of the dried residue.This difference in organic content is consistent with the appreciably greater oxygen uptake rate determined for the overburden material (Table 2). The metal content of each test sediment is given in Table 3.1bese values are presented only for sample "fingerprinting";and an analysis of metals dissolved (and conceivably biologically available)within freshwater suspensions of these test materials was beyond the terms of reference of this investigation. rL [ [ [ [ c D c c [ [ [ [ [ .[ l [1 [ [ [ c [ c [ [ [ [ [ -19 - Thus the extent to which a high concentration of certain constituents (i.e.the arsenic content of the paydirt sample)mayor may not contribute to any biological effects noted for freshwater suspensions of these materials cannot be ascertained from these data. Figures 7 and 8 illustrate the interrelationship of total residue, nonfiltrable residue and turbidity values for differing strengths of each sediment type suspended in freshwater.For both the inorganic and organic sediments,total and nonfiltrable residue values for each sample were almost identical.These values were highly correlated with sample turbidity,and with nominal concentrations (Fig.7 and 8). Results from these analyses indicate that the total residue values determined for suspensions of paydirt or overburden taken during the bioassay tests are good approximations of their total nonfiltrable residue (suspended solids)content. The turbidity values measured for each suspension allow rough calculations (based on total res idue determinations)of the turbidi ties of the test sus pens ions to which fish were exposed. The degree to which sediment loadings remained in suspension within the recycle test tanks,throughout a 96-h test period,is illustrated in Figure 9. Total residue values determined for the lowest paydirt strength monitored (500 mg -L -1)varied appreciably and showed a trend to decline with respect to time sampled;whereas respective values for the higher strengths monitored (5,000-100,000 mg-L-1)were more consistent throughout the 96-h test period.The total residue concentrations measured for each treatment were generally lower than the nominal (pre-weighed)strength of sediment added to each tank (Fig.9). Information concerning the dispersal pattern for suspended sediment within the recycle test tanks is provided in Table 4.For each suspension examined,the total residue value for the sample taken from the hose outlet (tank inflow)was appreciably (2-5 times)higher than respective values for samples taken from the surface,mid-depth or bottom locations within the net enclosures where fish were held.For paydirt strengths >5,000 mg °L -1,total residue values for the surface water were slightly but consistently lower than respective values for samples taken mid-depth or from the bottom of the net enclosure (Table 4).These results indicate that,for each of these test suspensions,a portion of the sediment fines was settling and being recirculated.Since the gradient for suspended solids within the portion of each tank to which fish were confined was small,and since these fish were continuously subjected to the settleable solids,the nominal (pre-weighed)strength of paydirt prepared in each recycle tank should approximate that to which the fish were exposed.However,this may not be true for the lower concentrations (i.e.<500 mgoL-l;Fig.9)tested in these tanks.In such instances a greater proportion of the sediment fines added may adhere to the netting or tank sides,and therefore be effectively unavailable. -20 - Acute survival and gill histology All lSoC~acclimated grayling survived a 4-day exposure to each suspended paydirt strength examined,up to and including 250,000 mgoL-1 (Table S). Additionally,the five grayling held in 50,000 mgoL-1 for a more prolonged period (16 days)survived this extended exposure.All hatchery-reared rainbow trout fry introduced to these suspensions at the termination of the bioassays with grayling also survived for 96 h. All grayling held in a 100,000 mgoL -1 strength of the sample of paydirt prepared by fine-sieving only (no pre-grinding of test material)survived the 4-day exposure.These fish were active and showed no overt signs of damage at this time.Particle size analysis of the sediment to which this group of fish was exposed indicated that 47%of the sample was coarse sand (>0.2 mm),and that only 22%of the test material was very fine particles (<38 urn).Thus,as with the pulverized preparations of paydirt fines examined,this coarser inorganic sediment suspension also permitted acute survival of underyearling grayling. All graylinf acclimated to lSoC and held in organic overburden suspensions up to SO,OOO mgoL-(nominal strength)survived the 4-day test period (Table 6). Those fish acclimated to SoC and exposed to coldwater suspensions of inorganic .paydirt fines up to and including 10,000 mgoL-1 also survived for four days; whereas mortalities of 10%(20,000 mgoL-1)to 20%(100,000 mgoL-l)were found with fish groups held in higher sediment strengths (Table 7).In these instances,fish deaths did not occur until after a 48-h exposure.All control fish examined in each survival test were alive and in apparently good condition throughout the 96-h period of exposure. Behavioural observations of fish during the acute survival tests were restricted due to the opacity of the suspensions.For the more dilute suspensions of paydirt or overburden permitting these observations (SO and 100 mg °L -1)'no signs of coughing or increased swimming activity were seen. In both the SoC and lSoC bioassays with paydirt,grayling held in sediment strengths >10,000 mgoL-1 remained at the surface of each suspension.These fish showed no-signs of respiratory distress.In exhi~iting this behavioural abnormality,the fish could have been detecting and;tseeking lower sediment strengths within the surficial waters (Table 4).Surfacing of fish was not apparent in the lower strengths of paydirt examined nor in any concentration of suspended overburden. Inspection of fish at time of autopsy indicated no overt signs of distress or damage (i.e.lethargy,fin or snout erosion,skin lesions,exophthalmos,external or internal bleeding).Gross examination of gills showed no increased mucous production nor signs of damaged tissue attributable to any strengths of paydirt (50 or lS o C tests)or overburden to which these fish were exposed for four days. No changes in gill his tology occurred due to exposure of any grayling to suspensions of paydirt up to and including 100,000 mgoL-l.The acute effect of higher sediment strengths (i.e.250,000 mgoL-1)on gill morphology was not [ F L [ [ [ [ [ [ [ [ [ [ r L [ L [ [~ [ [ [ c [.. [ [ [ [ l [. [ -21 - examined.Similarly,the appearance of the gill tissues examined for fish held in overburden suspensions of 0 (freshwater control)to 50,000 mg'L-1 for four days was identical. The structure of the gill filaments and lamellae of all tissues examined was,in general,similar to that described as "normal"for other salmonid fish species (Herbert and Merkens 1961;Morgan and Tovell 1969;Noggle 1978).Gill filaments were covered by a thick stratified epithelium,whereas the secondary lamellae consisted of leaf-life structures composed of a pillar cell system delineating blood capillaries covered by a monolayer of flattened or sl ightly enlarged epithelial cells. For the gill tissues examined,no hyperplasia,clubbing or fusion of lamellae was caused by any sediment exposures.Sediment particles were frequently observed between adjacent gill filaments of exposed fish.The amount of sediment observed appeared to be correlated with the concentration of suspended material to which fish were exposed.Otherwise,the histology of gills from fish held in suspensions of overburden or paydirt fines for four days could not be distinguished from that of the freshwater controls. Water quality conditions (pH,temperature,conductance,dissolved oxygen content)to which grayling were exposed during the 4-day survival tests are presented in Appendices 2-4.In each bioassay,the temperature of each suspension varied by 1 0 C or less throughout the test period.The dissolved oxygen content of each suspension was unaffected by sediment type or strength,and was maintained above 80%saturation by the recirculating test apparatus.The pH values of each suspension were also unaffected by sediment strength or type.For both the paydirt and overburden materials,the higher (~1,000 mg 'L-1)concentrations of suspensions examined caused a slight «20 umho 'cm-1 )but consistent elevation in conductivity.This minor difference was evident within 30 min of startup of each test,and did not change appreciably with respect to time. To the best of our knowledge,the lethal tolerance of Arctic grayling to suspended sediment has not been examined previously under controlled conditions. Available information concerning the lethal tolerance of other salmonid fish species to sediment is sparse and somewhat inconsistent.Smith (978)reported that high concentrations (28,000-55,000 mg 'L-l)of suspensions prepared from two natural sediment sources were required to cause mortalities of chum salmon (Oncorhynchus keta)fry within four days.On the other hand,Herbert and Merkens (961)reported that a 10-to 25-day exposure of juvenile rainbow trout to suspensions of kaolin or diatomaceous earth as low as 270 mg'L-1 caused significant mortalities of test fish.Noggle (1978)determined that the acute lethal tolerance (96-h LC50 values)to suspensions of natural sediments for groups of wild or hatchery-reared juvenile coho salmon,chinook salmon (0.tshawytscha) or steelhead trout (~.gairdneri)varied from 1,200 to 35,000 mg'L-T.Differences noted were attributed largely to seasonal temperature variations,with a lower tolerance of fish to sediment observed during the summer months.This conclusion was supported by a report of lower lethal concentrations of natural sediments for non-salmonid fish species,with higher test temperatures (Rogers 1969). -22 - The present findings for grayling indicate that this salmonid fish speCies can survive short-term exposure to very high concent~ations of suspended inorganic or organic sediment under controlled laboratory conditions,despite changes in season and temperature.The minor «20%)mortalities noted for grayling acclimated to SoC and held in 10,000 or-100,000 mg °L -1 strengths of coldwater (SOC)suspensions of inorganic sediment fines for four days suggest a decrease in lethal tolerance to sediment for fish acclimated to colder water;however, confirmation of this requires further studies.Since hatchery-reared rainbow trout swimup fry also survived acute exposure to very high strengths of the suspended inorganic fines to which grayling were exposed,one should not conclude that Arctic grayling are more tolerant to suspended sediment than other salmonid fish species.Perhaps differences in the nature of the suspended material examined in these versus previous (Herbert and Merkens 1961;Noggle 1978;Smith 1978)tests better explain the disparities in lethal tolerance to sediment noted. Assuming that other water quality conditions were compatible with fish survival, it is unlikely that suspended concentrations of the overburden muck or paydirt fines examined in these tests could be elevated sufficiently in natural streams to cause direct mortalities of resident populations of healthy juvenile Arctic grayling or other salmonid fish species due to short-term exposures. A number of investigators have found histopathological changes in fish gills attributable to sediment exposure.Herbert and Merkens (1961)observed thickening and fusion of secondary gill lamellae of some rainbow trout exposed for several weeks to diatomaceous earth or china clay.Noggle (1978)reported notable gill histopathologies in certain juvellile salmonid fish held in inorganic sediment suspensions ..s.13 ,000 mg °L -1 for ~Wto 96 h.Other researchers have also reported thickening and fusion of gilL lamellae in trout held in suspensions of diatomaceous earth for up to 96 h (Noggle 1978)~Unlike these findings,no gill histopathologies attributable to short-term exposure of grayling to suspensions of inorganic or organic sediment were noted in the present laboratory tests. Similarly,Smith (1978)found no damage to gills of juvenile chum salmon (0.keta) acutely exposed to high (up to 55,000 mgoL-1 )concentrations of suspended inorganic sediment. In his review of the effects of suspended sediment on fish,Pickral (1981) cited the variability in findings of fish gill tissue damage caused by high concentrations of suspended sediment,and suggested a lack of convincing evidence for such an effect.We interpret this variability in response to differing sediment characteristics (particle shape,size,hardness),biological (fish age, size and prior history of exposure)and experimental (exposure period,test apparatus)differences.More detailed examinations of gill histology for fish held in differing types of sediment sus pens ions under controlled conditions are required in order to understand the relevance of these variables. [ FL I [ [ [ [ [ c [ [ [ [ [ l Temperature tolerance tests Mean critical therm;l maxima (~pper lethal temperatures)for groups gf .[ grayling acclimated to 15 C water",and tested in freshwater only were 27.5-27.9 C and variances (SD)were small (Ta~Jes 8 and 9).Mean values for fish groups held in paydirt strengths of 500-100,000 mgoL-l were reduced slightly but consistently .[ [ [ [ [ c [ c [ c [ [ [ [ [. [ -23 - from control values (Table 8)and,at least for the higher sediment suspensions examined (>S,OOO mgoL-1 ),showed a somewhat greater response to increasing sediment strengths (Fig.10).Variances (SD,9S%confidence interval)for each group were unaffected by treatment.The highest paydirt strengths to which grayling were exposed (SO,OOO and 100,000 mgoL-l)reduced their mean critical thermal maxima by only l o C. For grayling acclimated to lS o C and 'held in various strengths of suspended overburden,temperatures at death were somewhat more variable and did not decline consistently from those for control fish until test concentrations exceeded a nominal strength of 1,000 mgoL-l (Table 9,Fig.11).Mean critical thermal maxima for groups of grayling held in overburden concentrations of S,OOO-SO,OOO mgoL-1 were decreased from the control value by only 0.2-0.7 o C. Exposure of these warmwater-acclimated grayling to solutions of the reference toxicant pentachlorophenol caused a more definitive response.Sublethal strengths of pentachlorophenol (0.4 and 0.7 of the 96-h LCSO value)reduced the critical thermal maxima for grayling by 0.9 and 1.8 C respectively;a strength equivalent to 1.2 LCSO caused a further reduction (Table 10).Variances (SD values)in temperature at death were also increased by this toxicant. Mean critical thermal maxima for the groups of grayling accl imated to SoC water and tested in freshwa ter only were 24.8-24.9 0 c.Upper lethal temperatures for these coldwater-adapted fish were not reduced by any yaydirt suspensions to which they were exposed,up to and including SO,OOO mgoL-(Table 11;Fig.12). Standard deviations calculated for each group (including control fish)were greater than any found for groups of grayling acclim~ted to lSoc. The median effective concentration (ECSO)of paydirt causing a net significant decline in critical thermal maxima for grayling acclimated to lSo C was 100 mgoL-l (9S%confidence interval,SO-SOO);whereas that derived for overburden was 8,471 mgoL-1 (1,S74->SO,000).No value could be calculated for the coldwater- acclimated grayling exposed to paydirt suspensions,as no response to sediment was observed. The upper lethal temperature tolerance for Arctic grayling,determined in this study for groups of fish held in clear freshwater only,was similar to values derived previously under identical procedures using underyearling coho salmon or rainbow trout (McLeay and Howard 1977;McLeay and Gordon 1980).LaPerriere and Carl son (1973)reported earlier,that the (high)thermal tolerance of various life stages of Arctic grayling was similar to other salmonid fish species.The increased resistance to high temperature with an increased temperature of acclimation (lSOC vs SOC)noted for grayling in the present study is also consistent with earlier findings for other species of salmonid fish (Brett 19S2; Black 19S3).TIle seasonal photoperiod to which fish are acclimated can also influence thermal tolerance (McLeay and Gordon 1978).TIlus differences noted in temperature tolerance of grayling acclimated to ISO or SoC probably reflect the effect of a number of variables (i.e.seasonal photoperiod,developmental stage of fish,fish condition)besides the temperature to which fish were acclimated. -24 - Sublethal concentrations of a number of aquatic contaminants (i.e.pulpmill effluent,herbicides,certain heavy metals)have been shown previously to cause a concentration-related decrease in the temperature tolerance of salmonid fish (McLeay and Gordon 1978,1980).The degree to which this tolerance is impaired is contaminant-specific;and findings to date indicate that contaminants which block oxygen exchange at the gills,or otherwise impair tissue respiration,cause a greater effect than those which exert their toxic effects in other ways (Wedemeyer and McLeay 1981). Since sublethal strengths of some aquatic contaminants can lower the upper lethal temperature tolerance of sa1monid fish by as much as 4-S o C (McLeay and Gordon 1978,1980),the minimal «l o C;lS o C fish)or negligible (SOC fish) responses caused by exposing grayling to very high concentrations of suspended inorganic or organic sediment indicate that these sediment loadings do not interfere to a large extent with the immediate thermal adaptive capacity of grayling.These findings,considered together with findings of thermal tolerance effects noted previously for sa1monid fish and other aquatic contaminants,suggest that short-term exposure of juvenile grayling to high loadings of suspended sediment may not impair their tissue respiration to a significant extent. Nevertheless,the threshold-effect (ECSO)levels of 100 mg e L-1 (paydirt)and 8,471 mg e L-1 (overburden)determined for the lSo C-acclimated grayling indicate that a measurable reduction in critical thermal maxima for these fish was caused by these and higher sediment strengths.The envirornnenta1 relevance of this response cannot be ascertained without further studies. The greater reduction in critical thermal maxima values for grayling exposed to pentachlorophenol in this study was consistent with that for other sa1monid fish species challenged with this reference toxicant (McLeay and Gordon 1980). This finding indicates that'the tolerance of grayling to temperature extremes is similarly influenced by this aquatic contaminant,and that the magnitude of effect is dependent on both the nature and concentrations of toxicant to which fish are subjected. Sealed jar bioassays Mean times to death for groups of warmwater-acclimated (lSOC)grayling held in jars containing various strengths of paydirt sediment increased progressively with concentration (Table 12,Fig.13).However,mean residual oxygen values for each treatment did not differ from control values (Table 12,Fig.14).A repeat of this bioassay test using paydirt suspensions and coldwater-accl imated (SOC) fish showed no effect of this inorganic sediment on either times to death or residual oxygen values at death of these fish (Table 13,Figs.IS and 16). Unlike these findings,times to death for groups of grayling held in jars containing suspensions of overburden decreased progressively with increasing sediment strength (Table 14,Fig.17).Residual oxygen values for each treatment were somewhat more variable than was found for fish held in paydirt,but showed no consistent change with respect to concentration (Table 14,Fig.18). Median effective concentrations (nominal)of paydirt or overburden which affected times to death of the lSoC-acclimated grayling were 4,407 mg'L-1 [ f [ [ [ [ c [ c [ [ [ [ [ [ [ [' n [ [ [ D o [ [ [ [ [ [. [~ [ -Z5 - (Z97-ZZ,933 mg °L-1)and 161 mg °L -1 (4-615 mgoL -1),respectively.Thresho1d-effec t concentrations could not be calculated for residual oxygen levels due to the absence of consistent responses of this variable in each of the sealed jar bioassays conducted. Mean test temperatures (ZO.0-ZO.5 0 C for 150 C-acclimated fish;9.Z-9.9 0 C for 5 0 c-acc1imated fish)were held near-constant in each bioassay (Tables 1Z-14). Concentrations of paydirt remaining in suspension at the end of each 30-min settling period were decreased approximately Z-to 3-fo1d from respective values for the freshly-agitated suspensions (Tables 1Z and 13).The overburden suspensions to which fish were exposed in sealed jar bioassays settled to an even greater extent (Table 14)prior to their re-suspension. Grayling exposed to pentachlorophenol in sealed jar bioassays showed a concentration-dependent increase in residual oxygen values,together with a a progressive decrease in times to death of fish (Table 15).Sublethal strengths of 35 ugoL-1 (0.5 of the 96-h LC50 value)and 50 ugoL-1 (0.7 LC50)elevated residual oxygen values from those determined for each of the control (freshwater) groups. The tolerance of the warmwater-acc1imated grayling to hypoxia (oxygen deficiency)was similar to that found previously for underyearling coho salmon Or rainbow trout under identical test conditions (McLeay 1976;Gordon and McLeay 1977).The critical residual dissolved oxygen level at which each of these fish species die,if acclimated to 15 0 C and held in freshwater at ZOOC,is approximately Z.O mg 0ZoL -1.The even greater tolerance to hypoxic conditions found in the present bioassays with gray1i~acclimated to 5 0 c and tested at 100 C (critical value approximately 1.5 mg 0ZoL-1 )is also consistent with findings for other sa1monid fish in response to a decrease in test temperature (Gordon and McLeay 1977).LaPerriere and Carlson (973)cited field observations of Arctic grayling under ice cover,where the dissolved oxygen concentration of the surrounding water approached 0 mgoL -1.The present findings do not suggest a greater capacity to adapt to hypoxic conditions for this species than has been determined previously for other sa1monid fish. The respiratory responses of grayling to pentachlorophenol (elevated residual dissolved oxygen values,decreased times to death)are consistent with the response to this reference toxicant noted for underyearling rainbow trout when tested under identical conditions (McLeay and Gordon 1980).This toxicant is known to increase the oxygen consumption rate of sa1monid fish,and is believed to uncouple mitochondrial respiration (Chapman and Shumway 1978). The progressive decline in times to death of grayl ing exposed to increasing strengths of overburden (Fig.17)is likely due to the high oxygen demand demonstrated for this organic sediment (Table Z),and does not reflect a respiratory response of the test fish.On the other hand,the increase in mean times to death for 150 c-acc1imated grayling,with increasing strengths of paydirt (Fig.13),does suggest a reduction in respiratory rate attributable to this inorganic sediment.This response could be due to increased swimming activity of fish in the clear solutions,in response to visual "disturbance"during the -26 - bioassay.Alternatively,it could reflect decreased physical activity,reduced ventilatory rate,or decreased efficiency of oxygen transfer,caused by progressively higher strengths of paydirt.The significance of this response is unclear in view of the lack of effect of these paydirt·suspensions on the fishes' tolerance to hypoxia (Fig.14),and on the absence of a time-to-death response to paydirt for grayling acclimated to cold water (Fig.15). The effect of suspended sediment on the respiration rate of fish has not been examined to any extent.Neumann et al.(975)reported no change in the respiratory rates of oyster toadfish (Opsanus tau)held briefly in a 2,000 mg'L-1 suspension of natural sediment;although a 72-h exposure to 11,000 mg'L-1 caused a greater variance in oxygen uptake rates compared with control fish. Aquatic contaminants known to affect fish respiration normally cause a concentration-dependent increase in residual oxygen levels of salmonid fish held in sealed jars (McLeay 1976;Vigers and Maynard 1977);whereas those contaminants known to exert their toxic effects otherwise may not effect this response (McLeay and Gordon 1980).The absence of significant changes in residual oxygen values for grayling held in suspensions of paydirt or overburden suggests either that these sediments do not impair their tolerance to hypoxic conditions,or that the strengths of sediment to which fish were exposed were too low to evoke a response.The elevated residual oxygen values for grayling held in sublethal strengths of pentachlorophenol confirm that these fish will indeed show a response in this bioassay test to a contaminant known to affect fish respiration.The absence of gill lesions associated with 4-day exposures of grayling to these sediments,together with the 1 imited-if-any effects of paydirt or overburden suspensions on their temperature tolerance,further support the suggestion that short-term exposure of underyearling grayling to high suspended loadings of either of these sediment types does not impact their respiratory capacity. Acute stress bioassays Hematocrit values for groups of warmwater-acclimated grayling held in differing strengths of paydirt or overburden for 24 h were unchanged from corresponding control values (Tables 16 and 17).However,mean leucocrit values for these fish were consistently decreased by exposure to nominal sediment strengths of 1,000 mg'L-1 and higher (Figs.19 and 20).Median effective concentrations of paydirt and overburden causing this response were 51,651 mg'L-1 (2,381-)100,000 mg'L-1)and 5,843 mg~L-1 (2,092-29,107 mg'L-1),respectively. Leucocrit values for the groups of control fish sampled at the beginning and end of these bioassay tests did not differ appreciably (means,1.2-1.4%;Tables 16 and 17)• Blood sugar values determined for these fish were also affected by sediment. Results for fish exposed to differing suspensions of paydirt were inconsistent. Generally,J:l!.eans and/or 95%confidence intervals for groups of fish exposed to paydirt strengths of 500 mg'L-lor higher were increased from control values (Fig.21);although blood sugar values for fish held in 5,000 mg'L-1 were unchanged from those for the final control group. [ ~rL I [ [ [ [ [ [ [ [ [ l ,[ [ [ [ [ [ [ c [ [ [ [ [ [, [. [ -27 - The elevated plasma glucose levels for the initial control group were atypical of the final controls and of other groups of control fish (Tables 16-18).These inconsistencies prevented the calculation of a threshold-effect concentration. Plasma glucose values for groups of fish exposed to suspensions of overburden were consistently elevated from those for control fish (Table 17;Fig.22).The median effective concentration of overburden causing this response was less than 50 mgoL-1. As with these acute stress bioassays,hematocrit values determined for the groups of warmwater-acc1imated grayling surviving a 4-day exposure to inorganic or organic suspensions were unchanged from values for controls (Tables 5 and 6). Mean hematocrit values derived for all groups of coldwater-acclimated grayling were again unaffected by sediment treatment;although these values were decreased consistently from those for the warmwater-acc1 ima ted fish (Table 7).Leucocrit values for all groups of coldwater-acclimated grayling examined (including control fish)were declined from values for'warmwater-acc1imated fish held in freshwater and were unchanged by treatment (Table 7). Plasma glucose levels for all groups of coldwater-acclimated grayling surviving a 4-day exposure to paydirt differed from those for the control group. Paydirt concentrations of 500 mgoL -1 and higher caused a consistent increase in sample means and 95%confidence intervals (Table 7;Fig.23). Fish hematocrit values generally are highly correlated with both erythrocyte (red blood cell)counts and blood hemoglobin content (Houston and DeWilde 1968, 1972).The decrease in hematocrit noted in this study for underyearling grayling acclimated to cold water has been reported previously for other species of sa1monid fish (Banks et a1.1971). Hypoxic conditions cause significant increases in hematocrit values for sa1monid fish (Ho1eton and Randall 1967;Swift and Lloyd 1974;Casillas and Smith 1977).However,changes in hematocrit values are somewhat resistant to acute stress,including that caused by exposure of fish to sublethal concentrations of a variety of aquatic contaminants (McLeay and Gordon 1977,1979,1980). As in the present studies,Noggle (1978)reported that hematocrit values of underyearling coho salmon were unchanged by holding fish for 96 h in suspended sediment concentrations equivalent to 0.8 of the 96-h LC50 value.Other studies with non-sa1monid fish species given short-term exposure to sediment have found unchanged,elevated or depressed hematocrit values (Berry 1973;Neumann et al. 1975). Unlike hematocrit values,1eucocrit values (or numbers of circulating 1eucocytes;i.e.white blood cells)for sa1monid fish can change rapidly and dramatically in response to stress (MCLeay 1975;McLeay and Gordon 1977;Wedemeyer and MCLeay 1981).Short-term exposure of rainbow trout or coho salmon to sublethal concentrations of aquatic contaminants as low as 0.1 of the 96-h LC50 can cause significant declines in these values,provided that test fish are in -28 - good condition and unstressed beforehand (McLeay and Howard 1977;McLeay and Gordon 1979,1980).The general decline in leucocrit values for warmwater- acclimated Arctic gray1 ing held in suspensions of paydirt or overburden for 24 h indicates that each of these sediment types was stressful to these fish.Values for control groups were similar to those found previously for underyearling coho salmon,and somewhat elevated from control values for rainbow trout (McLeay and Gordon 1977).The absence of a consistent 1eucocrit response for the coldwater-acclimated·grayling may reflect the influence of prior stress (i.e. disturbances to all control and test fish during the 4-day exposure period)or perhaps a differing mechanism of response to stress for co1dwater-versus warmwater-acc1imated fish. The stress reactions (depressed 1eucocrit values,elevated plasma glucose values)found for grayling exposed for 24 h to sublethal strengths of pentachlorophenol (Table 18)are typical of the acute responses shown previously to be elicited for underyearling rainbow trout by this toxicant (McLeay and Gordon 1980).Diverse environmental stressors including sublethal strengths of aquatic contaminants are known to cause a rapid elevation and/or increased variance in blood sugar levels for groups of sa1monid fish (McLeay 1977;Wedemeyer and McLeay 1981).A consideration of the blood sugar changes found in the present tests , together with the 1eucocrit changes for sediment-exposed fish,confirm that sublethal strengths of the paydirt and overburden suspensions examined were indeed acutely stressful to Arctic grayling.The variations in plasma glucose levels noted for coldwater-acclimated grayling exposed .tosuspensions of paydirt for 96 h indicate that grayling are stressed by this inorganic sediment,regardless of season or acclimation temperature. Other studies of the hematological effects of sediment suspensions on fish are limited.Noggle (1978)found that blood sugar values for goups of coho salmon held for 96 h in inorganic sediment suspensions >0.2 LC50 were significantly changed from those for control fish.Similarly,O'Connor et a1.(1977>reported that the laboratory exposure of a number of species of estuarine fish to suspensions of natural sediments caused hematological changes indicative of stress responses. General The threshold-effect concentrations (EC50)of paydirt or overburden suspensions calculated to cause acute responses for Arctic grayling in the present series of laboratory bioassays are stllD.marized in Table 19.The EC50 values presented are based on nominal (pre-weighed)strengths of sediment to which fish were exposed.From these data it can be seen that the acute lethal tolerance of both warmwater-and coldwater-acclimated grayling to suspensions of organic and/or inorganic sediment was too high to permit the calculation of LC50 values.For fish acclimated to l5 0 C,threshold-effect values were derived for some of the sublethal responses monitored (i.e.critical thermal maxima;leucocrit;time to death in sealed jar bioassays);whereas values could not be determined for other tests due to the absence of effect (i.e.residual oxygen)or increased variability of response (i.e.plasma glucose values for paydirt-exposed fish)caused by treatment.No threshold-effect values could be derived for fish acclimated to 5 0 C [ .rL, [ [ [ [ c c [ [ [ r L [ .[ [ r [ [ [ [ [ [ E c c c [ [ c. [~'.. .9' [ -29 - and challenged with differing suspensions of paydirt due to the absence (in the temperature tolerance and sealed jar bioassays)or increeased variability of responses (in the acute stress test)caused by sediment exposure. As with other hematological changes,variations in blood sugar values or blood cell counts (including 1eucocrit values)of fish in response to enviromnenta1 alterations reflect a nwnber of dynamic adaptive reactions.Such changes in regulatory precision of blood constituents may be evident by upward or downward shifts in values from the norm,although effects are also shown in many instances by increased variations in sample values from the normal range (B1axha11 1972;Wedemeyer and Nelson 1975).The present changes noted in blood sugar and 1eucocrit values of grayling due to suspensions of inorganic or organic sediment are consistent with these homeostatic effects.In such instances,the calculation of EC50 values based on a net significant increase (for blood sugar)or decrease (for 1eucocrit)of these values is inappropriate (Table 19).Derivation of meaningful threshold-effect concentrations for these tests would require the determination of normal ranges of these blood constituents for control fish,using a large sample size (n)200)(Wedemeyer and Nelson 1975).Individual values for each sediment-exposed fish would then be examined to determine if they were within this range. The present laboratory bioassays with inorganic and organic sediment indicate that these sediment types can cause dissimilar sublethal effects for Arctic grayling (i.e.time to death in sealed jar bioassay),or may differ appreciably in threshold-effect concentrations (Table 19).The nature of suspended sediment (including particle constituents,size and angularity),water velocity and other enviromnenta1 variables may contribute significantly to the manner and extent to which it causes adverse biological effects. FIELD STUDIES Water quali ty A swnmary of the results of all water quality sampling is presented in Tables 20 and 21.Detailed water quality data are presented in Appendices 5 through 9. The data indicate that turbidity and nonfi1trab1e residue were elevated in aighet Creek which was being mined,relative to Minto Creek upstream of the junction, during both August and September.Total volatile residue (organic material)was a small component of total residue in both streams.Turbidity in Highet Creek in August ha·d a mean value of 51 FTU and a range of 3 to 250 FTU,whereas in September mean turbidity was higher (636 FTU)and ranged from 100 to 2,250 FTU (Appendices 5 and 6).The mean turbidity level in Minto Creek during August was 1.1 FTll (range,0.7 to 1.8 FTU)and,in September,was 0.9 FTU (0.5 to 1.8 FTU). Nonfi1trab1e residue (suspended solids)reflected the same trends as turbidity for the two streams (Table 20),indicating the substantially higher load of suspended material being transported in Highet Creek as a result of placer mining activity upstream.Inclusion of dissolved solids with suspended solids (total residue)did not appreciably alter this similarity. -30 - Turbidity and total residue in Highet Creek during the two test periods showed similar trends and are illustrated in Figures 24 and 25.Turbidity and total residue were lower in Highet Creek in August than they were in September, reflecting differences in mining activity.The maximum total residue in August was 294 mg!L-l but in September reached 1,900 mg·L-I .It is apparent upon examination of Figs.24 and 25 that the suspended sediment concentration at the test site was consistently higher between 1800 hand 0800 h each day.The approximate 10-h delay between the daily onset of mining ("'0800 h)and the increase in sediment concentration was due to the travel time for transport of sediment between the test site and the mining operations,together with the retention time of the settling pond. Particle size analysis of suspended solids within Highet Creek is shown in Table 22.The distribution of particles was similar between the two test periods.These analyses showed that all solids were less than 400 urn in diameter.Solids between 2 and 50 urn in August comprised 83.9%of the material, and in September 88.7%of the material. Comparison of residue and turbidity values for Highet Creek water samples from inside and outs ide the net enclosures showed inconsistent results. Nonfiltrable residue concentrations in the grab samples were usually highest, averaging approximately 50%greater than automatic sampler values (Appendix 9). How-ever,total residue and turbidi ty levels for samples taken wi thin or outside of the cages showed much smaller differences. Dissolved oxygen remained near saturation in both streams during the two study periods (Table 20).Both Minto and Highet creeks exhibited circurnneutral pH,with Highet Creek ranging between 6.9 and 7.9 during the two study periods and Minto Creek ranging from 6.5 to 7.5 during the same time periods.Water temperature was also monitored and showed variability between the two sites. The temperature of Minto Creek water at the study site was relatively stable during each of the two test periods,ranging from 120 to 14°C in August and 50 to 7 0 C in September (Table 20).Temperature was probably maintained within these small ranges due to the stabilizing effect of Minto Lake,a few kilometres upsteam.Highet Creek exhibited the more usual temperature variation of free flowing streams which are more responsive to changing air temperatures.Water temperatures in this or other streams being mined may also be influenced through the use of the stream in removing frozen or cool temperature soils material. Temperatures in Highet Creek during August ranged from 7 0 to 120 C and during September,from 1 0 to 9 0 C (Table 20).The studies had been planned to coincide with warm water periods in August and cool water periods in September.This was achieved,although daily temperature regimes in the two streams were somewhat different. Several water quality characteristics for Minto and Highet creeks were determined at the beginning and conclusion of the two test periods.In addition, a single determination of the same characteristics was made on Mud Creek water,as some fish for the second test came from this creek.Results of all samplings are shown in Table 21.These data show the normal slight increase in hardness and [ ,r L I [ [ [ [ [ [ c c c [ [ [ l .[ [ [ [ [ c o c c [ [ c l c· [ -31 - alkalinity with increasing contribution of groundwater flows in late summer low flow periods.Hardness and alkalinity values were highest in Mud Creek (141 and 120 mgoL-1,respectively);slightly lower in Minto Creek (110 to 123 mgoL-l and 100 to 112 mgoL-1,respectively);and lowest in Highet Creek (73 to 80 mgoL-1 and 50 to 52 mgoL-1,respectively).As expected,calcium was the predominant anion in the three streams,ranging from 23.0 to 41.4 mgoL-l ,whereas magnesium ranged from 3.3 to 9.1 mgoL-l (Table 21). Analysis showed that most metals were either not different in concentration between streams,or were below the analytical detection limits (Table 21). Particular metals of concern Which were below detection limits at all times were lead,copper,mercury,and cadmium.Iron was found to be lower than detection limit in Mud Creek,and ranged from 0.1 to 0.5 mgoL-1 in Minto and Highet creeks. Strontium was found to be higher in Minto and Mud creeks (0.20 to 0.23 mgoL -1) than in Highet Creek (0.12 to 0.14 mgoL-1 ).Tin was above detection limit only during September in Highet Creek,when levels of 0.11 to 0.17 mgoL-l were reached.No guidelines have been established for the protection of aquatic biota for either strontium or tin (McNeely et al.1979). Four other metals (arsenic,manganese,zinc,and aluminum)were found to exceed recommended levels or tentative limits for the protection of aquatic biota on at least one occasion within the study area.Arsenic was above detection limits in Highet and Minto creeks only during August (Table 21).Levels in Minto Creek ranged from 0.01 to 0.07 mgoL-1 ,whereas in Highet Creek levels reached 0.1 to 0.2 mgoL-l •Arsenic levels defined as hazardous in the aquatic environment are 0.05 mgoL -1,or 0.01 mgoL -1 as presenting minimal risk to aquatic organisms (McNeely et ale 1979).Manganese was not detectable in Mud Creek or Minto Creek in September.However,manganese was found to be 0.014-0.016 mgoL-1 in Minto Creek during August,and,in Highet Creek,0.013 to 0.027 mgoL-1 in September and 0.048 to 0.052 mgoL-1 in August.Higher levels of manganese were found in August in the two study streams and in Highet Creek exceeded the recommended level of 0.02 mgoL -1 (McNeely et al.1979)in both periods (but only marginally in September).Zinc was detectable at all sites on all occasions (Table 21).Minto Creek zinc concentrations ranged from 0.02 to 0.03 mgoL -1,and Highet Creek concentations ranged from 0.02 to 0.05 mgoL-l.The recommended level of zinc for the protection of aquatic organisms is 0.03 mgoL -1 (McNeely et ale 1979),which is met or exceeded marginally in both study streams.Aluminum concentrations were found to be highest in August in both study streams and higher in Hi~het Creek than Minto Creek.Minto Creek levels ranged from <0.05 to 0.05 mgoL - ,whereas concentrations in Highet Creek ranged from 0.1 to 0.3 mgoL -1.No recommended limits for aquatic organism protection have been established for aluminum, although a tentative limit of 0.1 mgoL -1 has been identified (McNeely et al. 1979). As indicated,four metals were found to exceed recommended or tentative 1 imits for total metals derived for the aquatic environment.The metal values indicated in the present report are total levels;that is,all forms of the metal, whether combined with other elements,adsorbed to particles,or ionic.It is known that most metals are not acutely toxic to aquatic organisms in the non-ionic -32 - state,and that toxicity ~s not directly related to total metal values.In addition,at the pH and relatively high calcium levels found in the study area,it is expected that a large portion of the total metals measured would be in a non-ionic state. Caged fish studies Fish survival During the August tests,all grayling held for four days in cages within Highet Creek or Minto Creek were alive (Table 23)and in apparently good condition at the end of this exposure period.No signs of fish distress or injury were evident at either site.Similarly,all grayling captured from Minto Creek and held at the Highet and Minto Creek sites for five days during September 1982, survived.However,16%(five fish)of the Mud Creek fish held in Highet Creek during September died within 96 h (Table 23).All Mud Creek fish held in cages within Minto Creek survived the l20-h (5-day)test period. Although Mud Creek fish held in Minto Creek survived the test period while 5 of the 32 Mud Creek fish in Highet Creek died,it cannot be stated with certainty that suspended sediment (nonfiltrable residue)was the sole factor contributing to these fish deaths.Additionally,since all but one of these five fish survived the initial 48-h period following their transfer from Mud Creek (Table 23),the fish deaths were not caused by transfer shock alone.Perhaps these fish were unable to withstand the stress loading imposed upon them by the combined effects of capture,transport,confinement and the more rigorous environmental conditions within Highet Creek (daily fluctuations of water temperature to near zero, together with suspended sediment concentrations ~1,2l0 mgoL-l ). Our findings that wild Arctic grayling in stream environments can survive short-term exposure to suspended sediment concentrations of up to 1,210 mgoL-l are in general agreement with other field observations for this salmonid fish species.In a similar study,Simmons and LaPerriere (1982)found that underyearling Arctic grayling held for 7-10 days in the highly turbid ()1,000 NTU) water of Birch Creek,downstream of placer mining'activities,-survived. Atkins-Baker (MS,1980)also reported 100%survival of fingerling grayling held for 4 days in Hunker Creek when suspended sediment strengths ranged from 335 to 22,000 mg °L -1;although mortalities of caged fish were noted for other creeks in association with·peak loadings of suspended sediment.Mathers et al.(1981) captured adult grayling during July from other Yukon creeks (Clear,Duncan, Johnson)receiving placer mining effluent,at time~when suspended sediment concentrations varied from 114 to 4,453 mgoL-1 ,whereas fry were not found.These investigators did note,however,"On the other hand,good catches of grayling, both adults and fry,were obtained in Sulphur Creek.In this area suspended sediment concentrations were about 100 mgoL-l". Gill histology All gills of fish caged in Highet or Minto Creek during August or September appeared "normal"when inspected upon termination of these field bioassays.No c .n L [ [ [ [ [ [ c [ [ [ [ L .[ [ [ [ [ [ [ [ [ [ [ [ [ l [. [. l -33 - @\'~,:loss of red coloration was evident,and clogging of filaments with sediment particles was not observed.Additionally,no signs of increased mucous production were seen on either occasion for gills of fish caged in Highet Creek.Subsequent I\licroscopic examination of gill arches preserved with polyvinyl lactophenol also fevealed no damage to gill filaments of fish held at either site on these occasions •. The detailed examination of gill tissues from grayling held in cages within Minto or Highet Creek during September 1982 showed histopathological changes at each site.Whereas the structure of gill filaments and secondary lamellae for uncaged fish captured directly from Minto Creek at this time was normal in appearance (Fig.26),that of fish held captive in both Minto Creek and.Highet Creek was not (Fig.27).Moderate-to-marked hypertrophy (increase in cell size) and hyperplasia (increase in cell number)of the lamellar epithelium was evident for gills of all caged fish examined (Table 24).Additionally,the frequency of gill ectoparasites (tentatively identified as monogenetic trematodes)noted occasionally for three of the five uncaged upstream Minto Creek fish examined was increased appreciably for fish caged at each site (Figs.26 and 27;Table 24). With the exception of the infrequent observations of particulate debris (sediment) between gill filaments of fish caged in Highet Creek,no differences in gill histomorphologies for fish caged in Highet Creek versus Minto Creek were evident. Wobeser et a1.(1976)reported severe lamellar hyperplasia for gill tissues of captive Arctic grayling infested with large numbers of monogenetic trematodes.Birtwell et ale (1983)also found extensive gill histopathologies for adult Arctic grayling captured from Minto Creek downstream of its junction with Highet Creek.The present association of gill lesions with increased numbers of ectoparasites for caged grayling suggests that the histopathologies noted were caused by these parasites.Other investigators (Williams 1967;Kearns 1968)have reported a rapid buildup of gill parasites for wild fish held in captivity,as was found in this instance. The absence of any increased gill mucous production or of histopathological changes attributable to the suspended sediment loadings to which grayling were exposed in Highet Creek is consistent with our laboratory findings.Unlike these results,Simmons and LaPerriere (1982)observed increased gill mucous secretions for wild underyearling Arctic grayling held for 7-10 days in turbid water downstream of placer mining activity.In a separate field study,Herbert et ale (1961)reported gill lesions for trout captured from sediment-laden streams, whereas those taken from clearwater streams were normal.However,these differences may have resulted from prolonged exposure of fish to sediment.The conflicting reports from field or laboratory studies of the presence or absence of gill histopathologies attributable to exposure of fish to suspended sediment suggest that factors such as particle type (shape,size),sediment concentrations, and duration of fish exposure likely determine whether or not direct damage to gill tissue will occur. -34 - Hematology The hematocrit values determined for groups'of fish held in Minto Creek or Highet Creek during August and September were similar;and apparently unaffected by the differing water quality conditions noted for each site.On the other hand, plasma glucose values determined for the grayling sampled from Minto Creek cages' or directly from Mud Creek during August 1982 were similarly and consistently low; whereas mean glucose values for fish sampled from the Highet Creek cages at this time were elevated by approximately 30%(Table 25).Mean blood sugar values for grayling held in Highet Creek during September were increased from those for fish caged in Minto Creek by greater than 100%,regardless of fish source.These findings suggest that the water quality conditions to which grayling were exposed in Highet Creek,on each of the two occasions that the field bioassays were conducted,were more stressful to these fish than those within Minto Creek. Our findings from controlled laboratory bioassays with inorganic or organic sediments indicate that suspended sediment strengths wi thin the range of those found in Highet Creek during August or September can cause changes in blood sugar and leucocrit values,which typify short-term reactions to stressors.The relatively colder and more variable water temperatures for Highet Creek during August and September (Table 20)may also account for the site-specific differences in plasma glucose values noted on each occasion.During the extended period required for salmonid and other fish species to acclimate to cold water,a number of physiological stress responses normally occur,including the elevation of blood sugar levels (Schuh and Nace 1961;Nace and Schuh 1961;Allan 1971).The capture, transport,and confinement of fish can also cause appreciable changes in their blood sugar regulation (Silbergeld 1974;Hattingh 1976;McLeay 1977).Thus the elevated blood sugar values noted for grayling caged in Highet Creek likely reflect the combined influence of suspended sediment together with other stressors such as fluctuating water temperatures. CONCLUSIONS The present laboratory bioassays demonstrate that underyearling Arctic grayling can survive short-term exposure to very high levels ()50,000 mgoL-1)of suspended inorganic or organic sediment.Season (including acclimation temperature)does not cause any marked changes in this tolerance,although test results suggest a slight reduction in lethal tolerance to suspensions of inorganic paydirt for grayling acclimated to cold (5 0 c)water.These laboratory findings are consistent with the fish-survival data obtained for grayl ing held in turbid waters downstream of placer mining activity for 4 or 5 days. No gill histopathologies were found in either laboratory or field tests which could be attributed to acute exposure of grayling to sediment.Additionally,the laboratory bioassays indicated that the tolerance of grayling to hypoxic conditions or to upper lethal temperatures was not appreciably affected by suspensions of inorganic or organic sediment.The environmental significance of the slight but consistent decrease in critical thermal maxima for warmwater-acclimated grayling exposed to paydirt suspensions )100 mg °L -lor to [ f [ [ [ [ [ [ [ [ [ [ L .[ [ [ [ [ [ [ [ [ [ [ [ [ [ [. [, -35 - suspensions of overburden >8,000 mg·L-1 (based on EC50 values)is not known at the present time.Nor is the significance of the increased time to death (reduced oxygen uptake rate)for warmwater-acclimated grayling only,caused by paydirt suspensions >4,000 mg·L-1,understood.The finding of decreased times to death for grayling-held in sealed jars containing overburden concentrations >160 mg·L-1 is thought to reflect the oxygen demand of this organic muck.The environmental relevance'of this oxygen demand is site-specific,and would be modified markedly by factors such as overburden type and loading to receiving waters,flow "condi tions,water temperature and presence or absence of ice cover. The acute stress bioassays demonstrate that suspensions of both paydirt and overburden can be acutely stressful to underyearling Arctic grayling.Further the test results indicate that suspended sediment strengths as low as 50 mg·L-I (overburden)may be stressful to these fish;and that stress responses can be evoked for both co1dwater-and warmwater-acclimated fish.Differences in blood sugar values noted on each of two occasions for grayling caged in Highet versus Minto Creek also suggest that the Highet Creek site was more stressful to these fish.The environmental relevance of these responses to the immediate survival and long-term wellbeing of Arctic grayling cannot be ascertained without further studies.However,stressful conditions are well known to reduce the·adaptive responses of other salmonid fish species to natural environmental fluctuations, and to increase the susceptibility of fish to disease (Wedemeyer et ale 1976; Wedemeyer and McLeay 1981). Resul ts from the present laboratory and field studies should provide some direction for future investigations concerned with the acute or long-term biological effects of suspended sediment on resident fish species,and.with appropriate water quality objectives for sediment.The influence of sediment concentration and type (including particle shape and size)which causes stress responses in fish (and the environmental significance of these responses)deserves further attention,as does the impact of more prolonged exposures.Effects of sediment type and strength on fish behavioural responses (i.e.feeding, avoidance/preference reactions)and on other life stages are also largely unknown at present.A basic understanding of the degree to which these responses may be altered by sediment is essential before the biological relevance to Arctic grayling or other sensitive aquatic species of specific sediment loadings within natural waterbodies can be fully appreciated. ACKNOWLEDGEMENTS We wish to thank Mr.W.G.Whitley (Director,Yukon River Basin Study)for his support and technical advice during this study;Mr.H.F.McAlpine (Dept.Indian & Northern Affairs)for the provision of historical information and the ISCO samplers;and Messrs.S.Roxburgh (Fisheries &Oceans Canada),H.F.McAlpine and D.Davies for their help in site selection.The technical assistance of Dr.N. Lowes (gill histology),Mr.D.Bradley (fish maintenance),and the Environment Canada/Fisheries and Oceans laboratory at West Vancouver (water quality analyses) are gratefully acknowledged.Our thanks are also expressed to Mr.L.Hildebrand for his technical efforts during all aspects of the laboratory bioassays,to Mr. M.C.Nelson for his assistance in carrying out the field portion of the study, Mrs.S.C.Jones for drafting,and to Ms.L.Borleske for her typing and drafting skills. \ -36 - REFERENCES Alderdice,D.F.1963.Some effects of simultaneous variation in salinity, temperature and dissolved oxygen on the resistance of juvenile coho salmon (Oncorhynchus kisutch)to a toxic substance.Ph.D.Thesis.Univ.of Toronto.177 pp. Allan,G.D.1971.Measurement of plasma cortisol and histometry of the interrenal gland of juvenile pre-smolt coho salmon (Oncorhynchus kisutch Walbaum)during cold temperature acclimation.M.Sc.Thesis.Univ.of. British Columbia.63 pp. [ r'1~ [ Allen,W.T.R.and B.S.W. waterbodies in Canada. Cudbird.1971.Freeze-up and break-up Canadian Meteorological Service,Toronto. dates of [ Anon.1965.Water quality criteria for European finely divided solids and inland fisheries. Advisory Committee.Int.J.Air Wat.Poll.9: freshwater fish. European Inland 151-168. Report on Fisheries [ Anon.1972.Testing Sieves and Their Uses. Mentor,Ohio.48 pp. Handbook 53.W.S.Tyler Inc.[ Anon.1975.Sedimentation Engineering: American Society of Civil Engineers. Manual on Engineering Practice No.54. New York.745 pp.[ Anon.1979a.Environmental studies,Clear Creek gold dredlini,preliminary findings.Sigma Resources Consultants Ltd.Project report.Prepared for Queenstake Resources Ltd.29 pp. [ '.' Anon.1979b.Unpublished analyses of dredge spoils.B.C.Research.Vancouver.fJ Anon.1979c.Environmental Laboratory Manual.Department of Environment and Department of Fisheries and Oceans.West Vancouver. Anon.1980b.Historical Streamflow Summary.Yukon and Northwest Territories to 1979.Inland Waters Directorate,Environment Canada.Ottawa.96 pp. Atkins-Baker,S.1980.In situ bioassay study at a gold placer ml.nmg area in Yukon using Arctic grayling.Unpublished report.Environmental Protection Service.Whitehorse.9 pp +appendix. Water use technology for placer mining effluent control.Sigma Consultants Ltd.Proj.Rep.Prepared for Dept.Indian Northern 141 numb.pp. Anon.1980a. edition. Anon.1981. Resources Affairs. Standard Methods for the Examination of Water and Wastewater. Washington,D.C.1134 pp. 15th [ [ rL [ Banks,J.L.,L.G.Fowler and J.W.Elliott.1971.Effects of rearing temperature on growth,body form,and hema tology of fall chinook fingerlings.Progr. Fish Cult.33:20-26..[ Berry,C.R. killifish. 1973.The effect of resuspended Virginia J.Sci.24:120. estuarine sediment on the ,[ [ 1983. aquatic Can.MS. -37 - Biltwell,I.K.,G.Hartman,B.Anderson,D.J.McLeay and J.G.Malick. ,Brief examination of Arctic grayling (Thymallus arcticus)and invertebrates in the Minto Creek drainage,Mayo,Yukon Territory. Rept.Fish.Aquat.Sci.(In Prep.). Bishop,F.G.1971.Observations on spawning habits and fecundity of the Arctic grayling.Progr.Fish-Cult.33:12-19. Black,E.C. fishes. 1953.Upper lethal temperatures of some British Columbia freshwater J.Fish.Re's.Board Can.10:196-210. Blaxhall,P.C.1972.The haematological assessment of the health of freshwater fish.A review of selected literature.J.Fish BioI.4:593-604.[ Bostock,H.S.1939. 220,pp.7-9. Mining industry of Yukon,1938.Geol.Surv.Can.Memoir F""1, Brett,J.R.1952.Temperature tolerance Oncorhynchus.J.Fish.Res.Board Can.9: in young 265-323. Pacific salmon,genus u Cairnes,D.D.1915.Mayo area.Geol.Surv.Can.Summary Report for 1915,p.2. Car1ander,K.D.1969.Handbook of Freshwater Fishery Biology. Press,Ames.75 pp. Iowa State Univ. Casillas,E.and L.S.Smith.1977.Effect of stress on blood coagulation and haematology in rainbow trout (Salmo gairdneri).J.Fish BioI.10:481-491. Chapman,G.A.and D.L.Shwnway.1978.Effects of sodium pentachlorophenate.on survival and energy metabolism of embryonic and larval steelhead trout.pp. 285-299.In Pentachlorophenol:chemistry,pharmacology,and environmental toxico10gy.--K.R.Rao,ed.Plenum Press.New York. Davis,H.S.1967.Culture and Diseases of Game Fishes.Univ.of Calif.Pres •• Berkeley and Los Angeles.332 pp. Davis,J.C.and R.A.W.Hoos.1975.Use of sodium pentachlorophenate and dehydro- abietic acid as reference toxicants for salmonid bioassays.J.Fish.Res. Board Can.32:411-416. ~Emond,D.S.1982.Heavy minerals in the gravels of Highet Creek,Mayo Mining District,Yukon Territory.B.Sc.Thesis.Univ.of Ottawa.71 pp. Gordon,M.R.and D.J.McLeay.1977.Sealed-jar bioassays for pulpmill effluent toxicity:effects of fish species and temperature.J.Fish.Res.Board Can.34:1389-1396. Hattingh,J.1976. Labeo capensis. Blood sugar as an indicator of stress in the freshwater fish, J.Fish BioI.10:191-195. Herbert,D.W.M.andJ.C.Merkens.1961.The effect of suspended mineral solids on the survival of trout.Int.J.Air Wat.Poll.5:46-55. Herbert,D.W.M.,J.S.Alabaster,M.C.Dart and R.Lloyd.1961.The effect of china-clay wastes on trout streams.Int.J.Air Wat.Poll.5:56-74. -38 - c Holeton,G.F.and D.J.Randall.1967.The effect of hypoxia upon the partial pressure of gases in the blood and water afferent to the gills of rainbow trout.J.Exp.BioI.46:317-327. Horler,A.1980. Unpublished report. pp. Arctic grayling (Thymallus arcticus)investigations. Dept.Indian Northern Affairs.Whitehorse,Yukon.75 [ Houston,A.H.and M.A.DeWilde.1968.Hematological correlations in the rainbow trout,Salmo gairdneri.J.Fish.Res.Board Can.25:173-176. McLeay,D.J.and M.R.Gordon.1978.Effect of seasonal photoperiod on acute toxic responses of juvenile rainbow trout (Sa1mo gairdneri)to pu1pmill effluent.J.Fish.Res.Board Can.35:1388-1392. McLeay,D.J.1976.A rapid method for measuring the acute toxicity of pulpmill effluent and other toxicants to salmonid fish at ambient room temperature. J.Fish.Res.Board Can.33:1303-1311. Me Leay,D.J.1977.Development of a blood sugar bioassay for rapidly measuring stressful levels of pu1pmi11 effluent to salmonid fish.J.Fish.Res.Board Can.34:477-485. [ [ c c [ [ [ c c [ [ rL .... Summary and analysis of fish capture data from placer mining Can.MS.Rept.Fish.Aquat.Sci.(In Prep.) Knapp,W.1975. study,1975. LaPerriere,J.D.and R.F.Carlson.1973.Thermal tolerances of interior Alaskan Arctic grayling (Thymallus arcticus).Inst.Water Res.Rep.No.IWR-46. Univ.of Alaska,Fairbanks.36 pp. Kearns,G.C.1968.The development of the adhesive organism of some dip1ectamid, tetraonchid and dacty10grid gill parasites (Monogenes).Parasitology 58: 149-163. McLeay,D.J.and M.R.Gordon.1977.Leucocrit:a simple hematological technique for measuring acute stress in salmonid fish,including stressful concentrations of pulpmill effluent.J.Fish.Res.Board Can.34: 2164-2175. Houston,A.H.and M.A.DeWilde.1972.Some observations upon the relationship of microhaematocrit values to haemoglobin concentrations and erythrocyte numbers in the carp Cyprinus carpio L.and brook trout Sa1ve1inus fontina1is (Mitchil1).J.Fish Bio1.4:109-115. McLeay,D.J.1975.Sensitivity of blood cell counts in juvenile coho salmon (Oncorhynchus kisutch)to stressors including sublethal concentrations of pulpmi11 effluent and zinc.J.Fish.Res.Board Can.32:2357-2364. McLeay,D.J.and M.R.Gordon.1980.Short-term sublethal toxicity tests to assess safe levels of environmental contaminants.B.C.Research Proj.Report No.1-11-199.Prepared for Environment Canada.Vancouver.24 pp. McLeay,D.J.and T.E.Howard.1977.Comparison of rapid bioassay procedures for measuring toxic effects of bleached kraft mill effluent to fish.pp. 141-155.In Proc.3rd.Aquat.Tox.Workshop,Halifax,N.S.Nov.2-3,1976. Envir.Prot:"Servo Tech.Rep.No.EPS-5-AR-77-1.Halifax. ~[ [ -39 - McNeely,R.N.,V.P.Neimanis and L.Dwyer.1979.Water Quality Sourcebook.A guide to water quality parameters.Inland Waters Directorate,Environment Canada.Ottawa.89 pp. Mathers,J.S.,N.O.West,and B.Burns.1981.Aquatic and wildlife resources of seven Yukon streams subject to placer mining.Government of Canada, Departments of Fisheries and Oceans,Indian and Northern Affairs,and Environment.183 pp. Morgan,M.and P.W.A.Tovell.1969.The structure of the gill of the trout, Salmo gairdneri.Z.Zellforsch.142:147-162. Nace,P.F.and J.E.Schuh.1961.Environmental temperature change and blood ~sugar change in the toadfish,Opsanus tau.BioI.Bull.121:401. ~Neumann)D.A.,J.M.O'Connor,J.A.Sherk and K.V.Wood.1975.Respiratory and hematological responses of oyster toad fish (Opsanus tau)to suspended solids.Trans.Amer.Fish.Soc.104:775-781. Noggle,C.C.1978.Behavior,physiological and lethal effects of suspended sediment on juvenile salmonids.M.S.Thesis.Univ.Washington.87 pp. n J O'Connor,J.M.,D.A.Neumann and J.A.Sherk,Jr.1977.Sublethal effects of sus- pended sediments on estuarine fish.Maryland Univ.Solomons Nat.Res.lnst. Publ.No.CERC-TP-77-3.92 pp. Pickral,J.C.1981.State of the art of stream monitoring. Transportation Res.Counc.Report VHTRC 81-R34. Charlottesville,VA.52 pp. Virginia Highway & March 1981. Rawson,D.S.1950.The grayling (Thymallus signifer)in northern Saskatchewan. Can.Fish.Cult.6:3-10. '-"Rogers,B.A.1969.Tolerance levels of four species of estuarine fishes to sus- pended mineral solids.M.S.Thesis.Univ.Rhode Island.60 pp. Schmidt,D.and W.J.O'Brien.1982.Planktivorous feeding ecology of Arctic grayling (Thymallus arcticus).Can.J.Fish.Aquatic Sci.39:475-482. Schuh,J.E.and P.F.Nace.1961.Seasonal changes in blood sugar and related phenomena in the toadfish,Opsanus tau.BioI.Bull.121:406. Silbergeld,E.K.1974.Blood glucose:a sensitive indicator of environmental stress in fish.Bull.Environ.Contam.Toxicol.11:20-25. Simmons,R.and J.D.LaPerriere.1982.Unpublished findings.Alaska Cooperative Fishery Research Unit,Fairbanks. ,., I I I "L. [ Singleton,G.A.,O.A.Steen,K.Weagle and D.Weir.1981.Fish and wildlife habitat recovery in placer mined areas of the Yukon.Hardy Associates (1978) Ltd.Final report.Prepared for Department of Indian Affairs and Northern Development.131 pp. Smith,D.W.1978.Tolerance of juvenile chum salmon (Oncorhynchus keta)to sus- pended sediments.M.S.Thesis.Univ.Washington. -40 - [ Stephan,C.E.1977.Methods for calculating an LC50.pp.65-84.In Aquatic Toxicology and Hazard Evaluation.F.L.Mayer and J.L.Hamel ink ,eds.AS'lM STP 634.American Society for Testing and Materials.Philadelphia,PA. 1968.Avoidance reactions of rainbow trout to zinc sulphate Water Res.2:367-372. Sprague,J.B. solutions. Sprague,J.B.1973.The ABC's of pollutant bioassay using fish. Biological Methods for the Measurement of Water Quality. American Society for Testing Materials.Philadelphia,PA. pp.6-30.In AS'lM STP 528:'" r L I [ [ Swift,D.J.and R.Lloyd.1974.Changes in urine flow rate and hematocrit value of rainbow trout Salmo gairdneri (Richardson)exposed to hypoxia.J.Fish BioI.6:379-387.[ Tipper,H.W.,G.J.Woodsworth and H.Gabrielse. of the Canadian Cordillera and adjacent America.Geol.Surv.Can. 1982. parts Tectonics assemblage map of the United States of c Vigers,G.A.and A.W.Maynard.1977.The residual oxygen bioassay:a rapid pro-[, cedure to predict effluent toxicity to rainbow trout.Water Res.11: 343-346. Wallen,I.E.1951.The direct effects of turbidity on fishes.Bull.Okla.t Agric.Mech.ColI.,Arts Sci.Studies,BioI.Sere No.2.Vol 48 0):1-27. Weagle,K.1982..An evaluation of aquatic resources of Big Creek,Yukon Territory.Weagle Environmental Consultant Ltd.Project report.Prepared for Queenstake Resources Ltd.25 pp. Wedemeyer,G.A.and D.J.McLeay.1981.Methods for determining the tolerance of fishes to environmental stressors.pp.247-275.In Stress and Fish.A.D. Pickering,ed.Academic Press.London. Wedemeyer,G.A.and N.C.Nelson.1975.Statistical methods for estimating normal blood chemistry ranges and variance in rainbow trout (Salmo gairdneriY, Shasta strain.J.Fish.Res.Board Can.32:551-554. Wedemeyer,G.A.,F.P.Meyer and L.Smith.1976.Environmental stress and fish diseases.Book 5.pp.1-192.In Diseases of Fishes.S.F.Snieszko and H.R.Axelrod,eds.T.H.F.Publ.Inc.New Jersey. Weir,D.N.1979.An ecological reconnaissance of Klondike gold placer mining and discussion of interim guidelines for the issuing of authorizations to use water for the purposes of placer mining in the Yukon Territory.Prepared for the Klondike Miner's Association.45 pp. Williams,H.H.1967.Helminth diseases of fish.Helminthol.Abst.36:261-295. c [ [ [ [ [ [ Wobeser,G.,L.F.Kratt,R.J.F.Smith and G. branchiitis due to Tetraonchus rauschi Arctic grayling (Thymallus arcticus). 1817-1821. Ac omp an ad o. (Trematoda: J.Fish. 1976.Proliferative Monogenea)in captive Res.Board Can.33: ~[ [ [.,l J l J TABLE 1.Particle size distribution for paydirt and overburden sediment samples. Sieve Particle Paydirt (test no.1)Paydirt (test no.2)Overburden size size (mesh)(llm)weight cumulative weight cumulative weight cumulative ,,"- (%)we i'ght (%)weight (%)weight (%)(%)(%) +35 >400 0.0 0.0 0.0 0.0 31.0 31.0 +48 >300 0.0 0.0 0.1 0.1 9.5 40.5 +65 >210 0.2 0.2 0.7 0.8 8.9 49.4 .p- I- +100 >150 1.5 1.7 3.8 4.6 11.2 60.6 +150 >100 6.9 8.6 8.9 13.5 9.2 69.8 +200 >75 9.4 18.0 9.2 22.7 7.3 77 .1 +325 >45 9.8 27.8 9.4 32.1 8.6 85.7 +400 >38 6.8 34.6 7.5 39.6 11.0 96.7 -400 <38 65.4 100.0 60.4 100.0 3.3 100.0 -42 - TABLE 2.Moisture content,volatile and fixed residue,and oxygen uptake rate for paydirt and overburden sediment samples. [ [ I Characteristic Test No. Sediment type [ paydirt overburden 1 2.3 81.8 moisture content (%)2 2.8 84.6 3 2.5 82.5 volatile residue (%)1 3.6 96.0 fixed residue (%)1 96.4 4.0 0 1 0.01 0.08oxygenuptakerateat15C (mg 02 'ml-1 sediment-24 h-1 )2 0.01 0.08 0 1 0.01 0.6oxygenuptakerateat15C (mg °2-g-1 sedimenta -24 h-1 )2 0.01 0.6 aBased on dry weight. [ [ [ [ o [ [ [ [ [ [ .[ [ Metal contenta of paydirt and overburden sediment samples. Concentration (%dry weight) [ [ [ TABLE 3. Major components -43 - paydirt overburden Trace components Concentration (~g'g-l dry weight) aBased on analysis of sediment digest by inductively coupled argon plasma spectrograph. [ [ [ ... [ [ [ [ l [. [ alumina iron calcium magnesium sodium potassium antimony arsenic barium beryllium bismuth cadmium chromium cobalt copper lead manganese molybdenum nickel phosphorus silver strontium tin titanium vanadium zinc A120g Fe20g CaO MgO Na20 K20 Sb As Ba Be Bi Cd Cr Co Cu Pb Mn Mo Ni P04 Ag Sr Sn Ti V Zn 12.8 11.0 0.32 0.91 0.59 2.45 paydirt 73.8 1,570. 732. <0.3 <50. <1. 136. <5. 56.1 <10. 545. <4. 39.7 1,740. <0.5 83.5 <3. 3,090. 70.4 94.6 1.12 0.91 5.18 0.36 0.11 0.10 overburden <15. <30. 743. <0.3 <50. <1. 4.3 <5. 43.3 <10. 2,920. <4. 32.1 3,970. <0.5 156. <3. 229. 8.9 92.0 -44 - () [ r L aA1iquots (100 m1)taken for total residue analyses at 48 h after test sediment introduced to tanks. TABLE 4.Effect of location within recycle test tanks on co~centration (total residue values)a of recirculating paydirt sedimento Nominal paydirt strength (mgoL-1 ) 500 5,000 10,000 20,000 100,000 Total residue (mgoL-1 ) tank surface mid-depth bottom inflow (mid-tank)(mid-tank)(corner) 857 412 380 222 7,440 3,360 3,367 4,006 21,488 8,249 8,605 10,436 37,624 13,968 15,386 18,726 351,536 62,391 71,536 85,259 1~ [ [ [ [ [ [ ... [ [ [ [ [ [ [ .[ .[ [ J c,:]J J j I " TABLE 5.Acute survival test:Effect of a 4-day exposure to suspended inorganic paydirt fines on fish survival and on blood sugar and hematocrit values for underyearling Arctic graylinga acclimated to 15°C. Paydirt concentration Fish survival Plasma glucose Hematocrit (mgoL-1 )(%)(mg %)(%) .lb final 24h 48h 72h 96h Mean SD Mean SDnoml.na suspended residuec 0 0 100 100 100 100 129.8 59.1 30.6 1.9 50 76 100 100 100 100 104.2 25.5 31.6 2.8 100 261 100 100 100 100 132.2 31.3 31.0 1.6 1,000 1,090 100 100 100 100 97.6 20.8 30.4 2.1 I 5,000 5,401 100 100 100 100 65.6 5.1 33.2 2.3 .j::- VI 10,000 17,390 100 100 100 100 61.0 6 •.2 30.2 1.9 50,000 87,768 100 100 100 100 _d 100,000 237,959 100 100 100 100 56.6 7.5 32.6 4.7 250,000 100 100 100 100 ~Mean (±SD)weight,2.9 ±0.7 g;length,7.1 ±0.6 cm;condition factor,0.79 ±0.05. Based on weight of paydirt added to a 50-L test volume.C ''dBased on total residue for a 100-ml grab sample taken from the pump outlet at end of test. Not determined. TABLE 6.Acute survival test:Effect of a 4-day exposure to suspended organic overburden on fish survival and on blood sugar and hematocrit values for underyearling Arctic graylingaac- climated to 15°C. Overburden concentration Fish survival Plasma glucose Hematocrit (mgoL -1)(%)(mg %)(%) .lb final 24h 48h 72h 96h Hean SD Mean SDnoml.na suspended residuec I ~ 0 3 100 100 100 100 127.8 102.4 32.3 1.9 0\ . I 50 29 100 100 100 100 90.8 7.7 32.0 2.2 100 104 100 100 100 100 88.4 10.4 30.6 1.8 1,000 889 100 100 100 100 102.4 38.2 32.1 2.0 5,000 2,172 100 100 100 100 87.8 6.8 31.9 1.8 10,000 4,165 100 100 100 100 94.3 39.7 32.5 2.1 50,000 10,320 100 100 100 100 102.6 22.7 32.8 1.3 ~Mean (±SD)weight,3.6 ±1.0 g;length,7.8 ±0.7 cm;condition factor,0.75 ±0.04. Based on dry weight of overburden added to a 50-L test volume. cBased on total residue fora 100-mlgrab sample taken from centre of tank at end of test. r-l r--:: 1 TABLE 7.Acute survival test:Effect of a 4-day exposure to suspended inorganic paydirt fines on fish sur- vival and on blood sugar,leucocrit and hematocrit values for underyearling Arctic graylinga acclimated to 5°C. Paydirt concentration Fish survival Plasma glucose Hematocrit Leucocrit (mg.L-1)(%)(mg %)(%)(%) .lb final 24h 48h 72h 96h Mean SD Mean SD Mean SDnoml.na suspended ~ residuec 'l a 7 100 100 100 100 57.3 15.9 28.8 3.5 1.12 0.38 500 110 100 100 100 100 118.7 49.5 28.4 3.4 0.96 0.42 5,000 2,455 100 100 100 100 71.4 16.6 29.3 3.0 1.18 0.40 10,000 8,602 100 100 100 100 68.5 50.0 28.0 4.4 0.80 0.32 20,000 15,847 100.100 90 90 99.1 62.8 28.4 7.6 1.10 0.27 100,000 70,569 100 100 80 80 123.4 115.8 30.6 4.8 0.98 0.40 ~Mean (±SD)weight,4.5 ±1.3 g;length,8.4 ±0.8 cm;condition factor,0.76 ±0.09. Based on weight of paydirt added to a 50-L test volume. cBased on total residue for a 100-ml grab sample taken from centre of tank at end of test. -48 - TABLE 8.Temperature tolerance test:Effect of suspended inorganic paydirt on the critical thermal maxima for underyearling Arctic gray- linga acclimated to 15°C. a Mean (±SD)weight,5.1 ±0.9 g;length,8.3 ±0.4 cm;condition factor,0.90 ±0.06. bBased on weight of paydirt added to test volume. cBased on total residue for 100-m1 grab sample taken from centre of vessel at end of test. Temperature (OC)at death [ l~ I [ [ [ [ [ [ '- [ [ [0. [ [ [ SD 27.9 0.1 27.7 0.1 27.3·0.1 27.8 0.2 27.4 0.1 27.8 0.3 27.2 0.1 27.3 0.1 27.2 0.3 27.0 0.1 27.0 0.1 27.1 0.1 26.6 0.2 26.7 0.2 Mean Paydirt concentration (mg·L-I ) nomina1b final suspended residuec 0 (test 1)0 0 (test 2)0 25 28 50 76 100 (test 1)129 100 (test 2)85 500 490 1,000 (test 1)640 1,000 (test 2)780 5,000 3,410 10,000 6,200 20,000 11,000 50,000 61,272 100,000 82,275 L ~[ [ [-49·- r TABLE 9.Temperature tolerance test:Effect of suspended organic overburden on the critical thermal maxima for underyearling Arctic graylinga acclimated to 15°C. aMean (±SD)weight,4.8 ±1.0 g;length,8.1 ±0.7 cm;condition 8factor,0.91 ±0.07. aBased on dry weight of overburden added to test volume. Based on total residue for 100-ml grab sample taken from centre of vessel at end of test. Overburden concentration (rog'L-I ) 0 (control)a 100 125 150 147 500 609 1,000 700 5,000 2,336 10,000 3,646 20,000 7,242 50,000 14,197 Temperature (oC)at death SD 0.3 0.1 0.2 0.3 0.2 0.5 0.4 0.3 0.4 27.5 28.0 27.7 27.5 27.6 26.8 27.3 26.9 27.1 Meanfinal suspended residuec nominalb[ n [ [] ... C C C [ [ [ -50 - TABLE 10.Temperature tolerance test:Effect of pentachloro- phenol on the critical thermal maxima for underyear1ing Arctic gray1inga acclimated to 15°C. Pentachlorophenol Temperature (oC)at death concentration (l1g'L-1)Mean SD 0 (control)27.7 0.1 25 26.8 0.3 50 25.9 0.5 80 <25.5 a~ean (±SD)weight,4.9 ±0.9 g;length,8.2 ±0.5 cm; condition factor,0.89±0.06. [ [ [ [ [ [ .(. [ c [ [ [ l r1 . L [ -51 - TABLE 11.Temperature tolerance test:Effect of suspended inorganic paydirt on the critical thermal maxima for underyearling Arctic gray- linga acclimated to 5 °C. Paydirt concentration (mgoL-l ) 0'Temperature (eoC)at death 0 (control 1)3 0 (control 2)5 100 40 500 260. 1,000 520 10,000 5,692 50,000 71,143 [ n ! D i:lominalb final suspended residuec Mean 24.8 24.9 24.9 25;2 25.2 24.7 25.3 SD 1.5 1.4 1.5 2.1 0.8 1.1 1.3 r L [ [. a Mean (::tSD)w-eight,5.5 ±2.3 g;length,8.5 ±1.8 cm;'condition bfactor,0.81 ±0.12. Based on weight of paydirt added to test volume. cBased on total residue for 100-ml grab sample taken from centre of vessel at end of test. TABLE 12.Sealed jar bioassay:Effect of suspended inorganic paydirt on tolerance to hypoxia and time to hypoxic death for underyear1ing Arctic gray1inga acclimated to 15°C. Paydirt concentration Temperature at Time to death Residual oxygen at (mg.L-1)death (oC)(min)death (mg Oz·L-1) . 1 b initial final Mean SD Mean SD Mean SDnOTIl1.na suspended suspended residuec residued 0 <1 2 20.5 0.2 228 61 2.0 0.4 . 0 <1 1 20.5 0.1 224 58 1.9 0.2 . 100 107 52 20.5 0.1 220 58 1.9 0.2 U150038518020.5 0.1 270 60 1.9 0.3 N 2,500 1,836 420 20.5 0.1 245 73 1.9 0.3 10,000 8,127 2,670 20.3 0.1 271 71 1.9 0.3 20,000 15,483 5,997 20.4 0.1 2Q9 34 1.8 0.3 50,000 46,590 16,481 20.3 0.1 319 45 1.8 0.3 100,000 90,027 31,029 20.4 0.2 312 16 1.9 0.3 ?Mean (±SD)weight,10.4 ±1.3 g;length,10.2 ±0.4 cm;condition factor,0.98 ±0.07.~Based on weight of paydirt added to each of ten replicate 1.9-L glass jars. aBased on total residue for a 10D-ml grab sample taken immediately after the jar was inverted • ..Based on total residue for a 100-ml grab sample taken 30 min after the jar was inverted. TABLE 13.Sealed jar bioassay:Effect of suspended inorganic paydirt on tolerance to hypoxia and time to hypoxic death for underyear1ing Arctic gray1inga acclimated to 5°C. Paydirt concentration Temperature at Time'to death Residual oxygen at (mgoL-l )death (OC)(min)death (mg 02oL-I) . 1 b initial final Mean SD Mean SD Mean SDnoml.na suspended suspended residueC residued 0 6 6 9.4 0.6 491 63 1.5 0.4 0 4 5 9.7 1.0 498 94 1.6 0.4 100 66 24 9.2 1.5 452 96 1.6 0.2 500 194 81 9.6 0.6 456 87 1.7 0.5 2,500 1,699 402 9.8 0.4 438 95 1.4 0.3 U1 10,000 7,358 1,956 9.8 0.7 487 74 1.3 0.2 w 20,000 11,220 5,183 9.9 0.4 478 75 1.3 0.2 50,000 42,547 14,128 9.7 0.7 440 92 1.2 0.4 100,000 68,322 28,924 9.8 0.6 518 82 1.4 0.3 a 10.8 ±condition factor,0.88 ±0.06.bMean (±SD)weight,11.2 ±1.1 g;length,0.5 cm; Based on weight of paydirt added to each of ten replicate 1.9-L glass jars.CdBased on total residue for a 100-m1 grab sample taken immediately after the jar was inverted. Based on total residue for a 100-m1 grab sample taken 30 min after the jar was inverted. TABLE 14.Sealed jar bioassay:Effect of suspended organic overburden on tolerance to hypoxia and time to hypoxic death for underyear1ing Arctic gray1inga acclimated to 15°C. Overburden concentration Temperature at Time to death Residual oxygen at (mg.L-1)death cot)(min)death (mg 02·L-1) nomina1b initial final Mean SD Mean SD Mean SD suspended suspended residuec residued 0 <1 1 20.2 0.1 239 11 2.0 0.2 0 <1 2 20.3 0.1 227 40 2.0 0.3 100 120 43 20.3 0.1 209 44 2.1 0.3 VI.p. 1,000 786 347 20.3 0.1 189 22 2.3 0.4 5,000 4,123 832 20.3 0.1 203 16 1.8 0.3 10,000 7,870 941 20.4 0.1 187 35 2.1 0.6 20,000 12,870 1,027 20.3 0.1 175 36 2.1 0.4 50,000 23,200 2,723 20.0 0.1 136 24 2.3 0.6 a . bMean (±SD)weight,10.3 ±1.1 g;length,9.9 ±0.5 cm condition factor,0.96 ±0.07. Based on dry weight of overburden added to each of ten replicate l.9-L glass jars.ca:ased on total residue for a 100-ml grab sample taken immediately after the jar was inverted. ased on total residue for a 100-ml grab sample taken 30 min after the jar was inverted. r--,,) _..... .J E--J [..~.~L ...J L J ))L..;j TABLE 15.Sealed jar bioassay:Effect of pentachloropheno~on tolerance.to hypoxia and time to hypoxic death for underyearling Arctic graylinga.acclimated to 15°C. a Mean (±SD)weight,10.3 ±1.1 g;length,10.1 ±0.4 cm;condition factor, 0.98 ±0.07. TABLE 16.Acute stress bioassay:Effect of suspended inorganic paydirt on blood sugar,hematocrit and leuco- crit values for underyearling Arctic graylinga acclimated to 15°C. Paydirt concentration Plasma glucose Hematocrit Leucocrit (mg·L-I )(mg %)(%)(%) nominalb initial final Mean SD Mean SD Mean SD suspended suspended residuec residued 0 (initial 5 8 105.8 23.7 32.4 1.4 1.18 0.22control) 0 (final 2 5 82.5 7.7 33.9 2.4 1.29 0.16control) 50 43 44 91.3 20.7 35.1 2.6 1.35 0.24 100 102 85 98.8 12.3 34.4 3.1 1.22 0.27 \JI 90.8 28.4 33.7 1.8 1.21 0.29 (J"I500281402 1,000 420 560 120.9 11.0 34.3 2.9 1.13 0.26 5,000 4,121 7,650 82.3 10.1 34.8 3.4 0.96 0.16 10,000 15,725 10,230 116.2 54.6 33.4 2.5 1.07 0.37 20,000 17,135 22,840 73.1 26.1 34.4 4.9 0.93 0.28 100,000 56,960 118,305 102.6 32.2 34.7 4.6 1.09 0.47 a bMean (±SD)weight,3.8 ±0.7 g;length,7.9 ±0.6 cm;condition factor,0.76 ±0.05. cBased on weight of paydirt added to a 50-L test volume. Based on total residue for a 10o-L grab sample taken from centre of vessel at 0.5 h after ~ediment is added. Based on total residue for a 100-ml grab sample taken from centre of vessel at end of test. TABLE 17.Acute stress bioassay:Effect of suspended organic overburden on blood sugar,hemato-ocritand1eucocritvaluesforunderyear1ingArcticgray1ingaacclimatedto15 c. Overburden concentration Plasma glucose Hematocrit Leucocrit (mg·L-I )(mg %)(%)(%) . 1 b final Mean SD Mean SD Mean SDnoml.na suspended residuec 0 (initial 3 67.4 5.7 31.0 1.6 1.25 0.34 VIcontrol)'" 0 (final 5 74.4 10.2 32.0 2.2 1.36 0.46control) 50 30 94.5 12.8 33.1 0.6 1.22 0.39 100 190 85.8 13.5 31.6 2.0 1.35 0.13 1,000 1,590 101.2 10.1 33.2 2.0 1.15 0.13 5,000 2,757 80.9 10'.4 32.8 2.1 1.15 0.14 10,000 4,696 103.3 28.0 31.4 2.5 0.95 0.29 20,000 12,296 88.2 21..9 31.3 1.6 0.94 0.33 ~ean (±SD)weight,3.2 i:0.8 gj length,7.5 i:0.6 cm;condition factor,0.78 i:0.06. bBased on dry weight of overburden added to test volume. aBased on total residue for a lOO-r grab sample taken fro1ll.centre of vessel at end of test. TABLE 18.Acute stress bioassay:Effect of pentachlorophenol on blood sugar,hemato- crit and1eucocrit values for underyear1ing Arctic gray1inga acclimated to 15°C. a Mean (±SD)weight,3.3 ±0.6 g;length,7.6 ±0.7 cm;condition factor,0.75 ±0.06 • .' r-J TABLE 19.Summary of threshold-effect concentrations of paydirt or overburden suspensions causing acute re- sponses for Arctic grayling. Bioassay test temperature tolerance sealed jar bioassay sealed jar bioassay Acclimation temperature (OC) 15 15 15 15 Exposure (h) 96 12 5 5 Response decreased fish survival decreased critical thermal maxima increased (paydirt)or decreased (overburden)time to death increased residual oxygen at death paydirt >100,000 100 (50-500) 4,407 (297-22,933) >100,000 overburden >50,000 8,471 (1,574->50,000) 161 (4,...615) >50,000 1eucocrit stress test blood sugar stress test 15 15 24 24 decreased 1eucocrit values increased plasma glucose values 51,651 5,843 (2,381->100,000)(2,092-29,107) <50 LC50 temperature tolerance sealed jar bioassay sealed jar bioassay 1eucocrit stress testd dbloodsugarstresstest 5 5 5 5 5 5 96 20 8 8 96 96 decreased fish survival decreased critical thermal maxima increased time to death increased residual oxygen at death decreased 1eucocrit values increased plasma glucose values >100,000 >50,000 >100,000 >100,000 C C e e e e e e ~edian effective concentration causing a net significant response for 50%of fish (95%confidence interval bin parentheses). Median lethal concentration. cUnab1e to calculate due to increased variance of data. dNot conducted as a stress bioassay (values based on those for fish surviving a 96-h exposure).e .Not mea.sured. Table 200 Water quality characteristics monitored at test site in Highet Creek and the control site in Minto Creek during the fish enclosure tests,August and Septemeber,19820 range Highet Creek Minto Creek August 1982 September 1982 August 1982 September 1982 9.0 4.8 12.8 6.3 1.8 (9)2.7(7)0.7(7)1.3 (7) 7.0 -12.0 1.0 -9.0 12.0 -14.0 5.0 -7.0 11.0 12.6 10.2 10.2 0.7(9)0.9(7)0.7(7)0.5(7) 9.7 -11.5 9.5 -13.5 9.4 -11.0 10.0 -10.7 7.4 7.2 7.3 6.7 0.3(9)0.2 (7)0.2(7)0.2(7)0'a7.1 -7.9 6.9 -7.4 7.2 -7.5 6.5 -6.9 51 636 1.1 0.9 52 ~~O)483 (60)0.3(48)0.3(44) 3 -250 100 -2250 0.7 -1.8 0.5 -1.8 61 421 22 10 46 (78)257 (60)34(48)7 (44) <20 -208 80 -1210 <20 -40 <5 -34 161 637 152 54 (78)354 (60)47 (48) 79 -294 189 -1900 122 -319 146 585 119 52 (78)342 (60)37 (48) 77 -270 171 -1800 92 -246 18.0 47 35 8.0 (78)20 (60)13 (48) <10 -40 18 -110 19 -73 mean SD2 range mean SD range SD range mean SD range mean SD range Statistic pH Variable turbidity (FTU) total residue (mg·L-I ) water temperature (OC)1 dissolved dxygen (mgoL-I ) total fixed residue (mg·L-I )·mean SD range total volatile residue (mgoL-I)mean SD mean SD range nonfi1trab1e residue (mg·L-I )mean ITemperatures recorded by Birtwe11 et ale (1983)for the same August period were 6.0-9.5 in Highet Creek and 13.0-14.5 in Minto Creek. 2Number in brackets indicates number of samples analysed. .. r-1 r-:-;r--J r-J r-1 r-J r-J r-J r:-J''''c-J j r::J c-J c-J [""""'""]r-l rJ :-1 ~tT'J [-]L J l.J .J L J I L.J L..l ..J TABLE 21.Hardness,alkalinity and metal contenta (mgoL-I·)determined for water samples b taken from Highet,Minto and Mud creeks during the fish enclosure tests,August and September,1982. Variable Highet Creek Minto Creek Mud Creek August September August September September EDTA hardness C 73-75 79.,...80 110-114 123 141 a1ka1ini ty C 50-52 51 100-104 112 120 arsenic (As)0.1-0.2 <0.05 0.01-0.07 <0.05 <0.05 boron (B)<0.001 <0.001 <0.001 <0.001 0.011 barium (Ba)0.047-0.049 0.041-0.049 0.066-0.069 0.068 0.094 cadmium (Cd)<0.002 <0.002 <0.002 <0.002 <0.002 chromium (Cr)<0.005 <0.005 <0.005 <0.005 <0.005 copper (Cu)<0.005 <0.005 <0.005 <0.005 <0.005 mercury (Hg)<0.0002 <0.0002 <0.0002 <0.0002 <0.0002 manganese (Mn)0.048-0.052 0.013-0.027 0.014-0.016 <0.001 <0.001 0\ ~ nickel (Ni)<0.02 <0.02 <0.02 <0.02 <0.02 lead (Pb)<0.02 <0.02 <0.02 <0.02 <0.02 tin (Sb)<0.05 0.11-0.17 <0.05 <0.05 <0.05 strontium (Sr)0.12-0.13 0.14 0.21 0.23 0.20 zinc (Zn)0.05 0.02 0.02-0.03 0.02 0.01 aluminum (AI)0.2-0.3 0.1 <0.05-0.06 <0.05 <0.05 iron (Fe)0.3-0.5 0.1 0.4-0.5 0.4 <0.01 silicon (Si)4.2-4.4 4.3-4.4 2.9-3.0 3.2 2.4 calcium (Ca)23.0 25.4-26.0 31.3-32.4 36.1 41.4 magnesium (Mg)3.3-3.4 3.5-3.6 7.3-7.5 8.0 9.1 sodium (Na)1.6 1.9 1.8 2.1 1.4 br 0tal metal concentration,based on inductively coupled argon plasma spectrographic analysiso 4 samples were analysed for each trip and study site except Mud Creek where a single sample was co11ec~ef 0 amg CaC03·L·0 -62 - TABLE 22.Particle size distribution for suspended sediment sampled from Highet Creek during August and September,1982. Particle August 1982 suspensions September 1982 suspensions si2:ea weight cumulative weight cumulative.(Jm)(%)weight (%)(%)weight (%) >400 0 0 0 0 >50 1.5 1.5 0.2 0.2 >25 54.7 56.2 61.0 61.2 >2 29.2 85.4 27.7 88.9 <2 14.6 100.0 11.1 100.0 a the pipet method (Anon.1975).Measured using - rL [ [ [ [ [ [ [ "[ [ [ [ [ [ [ r~ I.~ TABLE 23.Percentage survival of underyear1ing Arctic grayling held in Highet Creek or Minto Creek for 4-5 days during August or September,1982. Test site Test period No.of Source Length Weight Fish survival (%) fish of fish (cm)a (g)a 24h 48h 72h 96h 120h Highet Creek August 5-9 99 Minto Creek 5.3 1.3 100 100 100 100b (0.7)(0.6) Minto Creek August 5-9 92 Minto Creek 5.2 1.3 100 100 100 100b (0.8)(0.5) Highet Creek September 10-15 36 Minto Creek 7.4 3.6 100 100 100 100 100 (1.3)(1.9) 0'1 UJ Highet Creek September 10-15 32 Mud Creek 6.5 2.4 97 97 84 84 84 (1.1)(1.7) Minto Creek September 10-15 35 Minto Creek 8.1 4.5 100 100 100 100 100 (1.4)(2.3) Minto Creek September 10-15 28 Mud Creek 6.5 2.3 100 100 100 100 100 (0.5)(0.7) abMean (±SD)values,measured at the termination of the exposure period. Experiment terminated at 96 h. TABLE 2.4.Gill histopatho1ogiesa for underyear1ing Arctic grayling held in Minto Creek or Highet Creek during September 1982. Treatment Fish b Hyperplasiac Clubbingd Debris eHypertrophyParasites no. caged in Minto Creek 1 +H--1+-1+f -1+ for 4 days 2 -1+++-1+ 3 -1++H--1++H- 4 +H--1+-1+-1+ 5 -1+-1++-1+ '"~ caged in Highet Creek 6 +H--1+-1+++H-I for 4 days 7 -1+-1+-1++-1+ 8 -1+-1++-1+-1+ 9 -1+-1+-1++-1+ 10 +H--1+++-1+ seined from Minto 11 ++++ Creek 12 +++ 13 -1++ apositive values are based on a scale of 1 to 4,where +=slight;-1+=moderate;+H-marked; band -1+-1+=very marked. Increase in cellular size. ~Increase in cellular number. Thickening of distal ends of lamellae. ;Tentative1y identified as monogenetic trematodes. Not evident. .. 1'""---' l.J ·-1 , I TABLE 25.Mean (±SD)biological characteristics of underyear1ing Arctic grayling sampled from cages or directly from creeks during August and September,1982. No.(n)and source Date Treatment Length Weight Condition Hematocrit Plasma of fish sampled (cm)(g)factor (%)glucose (K)(mg %) Mud Creek (19)09/08/82 seined from creek 5.9±0.6 1.9±0.5 0.88±0.06 43.6±4.9 63.9±8.9 Minto Creek (20)10/08/82 caged in Minto Creek 6.0±0.5 1.7±0.5 0.76±0.08 46.8±4.3 60.7±9.5 for 4 days Minto Creek (20)10/08/82 caged in Highet Creek 6.1±0.6 1.8±0.5 0.81±0.07 46.1±5.6 81.2±8.8 for 4 days Minto Creek (30)14/09/82 seined from creek 7.8±1.0 4.2±1.7 0.86±0.08 44.7±4.3 95.2±19.1 c:"\ Mud Creek (10)15/09/82 \...'1 caged in Minto Creek 6.7±0.5 2.4±0.6 0.79±0.07 46.1±4.5 64.1±12.0 for 4 days Minto Creek (10)15/09/82 caged in Minto Creek 8.9±1.7 5.9±3.0 0.76±0.05 55.6±2.5 132.4±54.9 for 4 days Mud Creek (10)15/09/82 caged in Highet Creek 7.4±1.5 3.6±2.8 0.77±0.03 50.6±3.7 159.3±69.2 for 4 days Minto Creek (10)15/09/82 caged in Highet Creek 9.1±0.6 6.3±1.5 0.82±0.05 52.6±4.3 276.4±141.4 for 4 days E u ID .., l i E u on .0 N I I T .... 67 PUMP (10 L·min-1) FIG. 1. Illustration of recycle test BOTTOM OF NYLON MESH CAGE INSERT tanks. 75mm RED BLOOD CELLS ------SEA LANT ~--BUFFY LAYER (WHITE BLOOD CELLS AND THROMBOCYTES) LEUCOCRIT (%)=HEIGHT OF BUFFY LAYER X 100 HEIGHT OF TOTAL BLOOD VOLUME PLASMA OR SERUM HEIGHT OF PACKED CELLSHEMATOCRIT(%)=--------------X 100 HEIGHT OF TOTAL BLOOD VOLUME TOTAL PACKED CELLS- FIG.2.Illustration of derivation of hematocrit and leucocrit values from a centrifuged blood sample within a heparinized glass capillary tube. TOTAL BLOOD VOLUME- Enclosure Settling Ponds t.:.-=-:..O====-2 Km FIG.3.Map of site for in-situ caged fish studies. [' [ -73 - heavy gauge wire hook draw string ---.----ttt---------,~~ melal post 4cm diameter ,-' L. [ [ [ [ [ [ [ [ 1.8m O.84m O.30m -- --~-''-'--''1 wire ring IW .'"..... water depth =O.5m r~ L. [ [. ['. , l PLAN VIEW posf'---__iIIIi opening fIG.4.Schematic drawing of net enclosures for in-situ caged fish studies. -75 - FIG. 5. Study site at Minto Creek. Fish enclosures are shown in-situ. FIG. 6. Study site at Highet Creek. Fish enclosures are shown in-situ. FIG.7.Relationship of total reSidue,nonfiltrable residue and turbidity for suspensions of paydirt sediment in freshwater. -77 - -i C ::0m o -i-<-"-i C- 00 0.000 .000 00.000 200 50 2.000 500 20.000 ~.ooo ~o.ooo 0--0 TOTAL RESIDUE .---.NONF ILTRABLE RES I DUE -------TURBIDITY 20 5 o 20 50 100 200 ~OO 1.000 2.000 ~.ooo 10.000 20.000 50.000 100.000 NOMINAL PAYDIRT CONCENTRATION <mg·t l ) o ~-1 ... ,r // .~~1 , 14"/ ~//// /V I I'/ r .F / .J ;I / // oJ V"/:l:'"'"I II / /1 .:/ ,,'/./ /~~ OI!~..-..;~~ ~ " 50 100 200 ~oo 20 5 o 1.000 2.000 ~.ooo 10.000 ~o.ooo 100.000 -'j'20.000 :...J•CI-E T...J -•WCI E :::>-0 W C/) :::>w 0 a::: C/)wwa:::...Jm ...J <{ ~a::: ~0....I.L. Z 0 Z rL [ c- [~ L [ [ [ [ [ [ n c o "c c c [ [ FIG.8.Relationship of total residue,nonfiltrable residue and turbidity for suspension~of overburden sediment in freshwater. -79 - -'TI c!- 00 0.000 200 5.000 50 20.000 -I C 2.000 ::0 CD .000 0 ~ 100.000 0---0 TOTAL RESIDUE e---e NONFILTRABLE RESIDUE • •TURBIDITY 20 5 o 20 50 100 200 500 1.000 2,000 5,000 10,000 20,000 50,000 100,000 NOMINAL OVERBURDEN CONCENTRATION <mge[l) i\ / / ~'/I I / "/ /1/ 7 / /.,/~ I ,/ (~/ ./v",;/ ~~/A'," "f'.I "/ "/ I'.// "/// I''/"~"".. ~,/ " 50 20 5 00 100 200 500 1.000 5.000 2.000 10,000 20.000 50.000 100.000 -I...J e 01-E~-e UJ01 E ::>-0 UJ en UJ::>0:0 CJ)UJ UJ ..J 0:m ..J <t 0:~~0 iL:F-Z 0 Z [ [' [~ [ [ [ [ c c '"[ .> C C [ [ [ [ c- [~ [ I...J 10,000 10,000 mg'l-l•0-0'0-A>-D----~0 E -0=- 1J.J:::> 5,000 o3'g'l-ICI0-.-..0-J'~Cf)~ 1J.J 0::: ...J j:! 0 1,000r- [ [ [ [ [ [ D c [ [ 100,000 100 -81 - t:'100,00~---------------eo-----o-_----=<>=<>0-__;...----=-e>= [ [ o 0.5 ••I ....__-'-L.'L.'.....'.T 5 24 48 72 96 TIME (h) [ L- [- l FIG.9.Illustration of the stability of differing concentra- tions of suspended paydirt fines within recycle test tanks during a 96-h bioassay. FIG.10.Relationship of concentration of suspended inorganic paydirt to critical thermal maxima for underyear1ing Arctic grayling acclimated to 15°C. !I I !I I "I , I ,!!" Note:Points represent mean; bars,95%confidence interval for 10 fish. !!!!I.II 50 100 1,000 10,000 100,000 NOMINAL PAYDIRT CONCENTRATION (mg-L-') I !!!!!!!I 29.0 test test----28.0 I 2 I 2 On 2 Q test-----I 2 :0 :0 :0 ~2 Q Q .027.0 ~Q 26.0 -83 - 23.0 :r: ~wo ~ W 0:: :::>~25.0 0:: Wa..r5 24.0 I- -0 0- [ [ [ [ [ [ [ c u ...c [ F...•.U [ [ [ [ [- [. [ 28.0 :0:2-2 2 Q £oU- J:27.0 ""~2~control w 0 ~26.0 w Ci:Note:Points represent mean;::::>25.0~bars,95%confidence interval Ci:for 10 fish. wa..24.0~w I- -85 - !! I !!'"!!! !!!"I !!I ,!" a.........,'1 1111,,1, 50 100 1,000 10,000 NOMINAL OVERBURDEN CONCENTRATION (dry weight basis) 23.0 29.0 [ [ [' [ [ [ [ [ c ... [ [ [j [ [ r~ L FIG.11.Relationship of concentration of suspended organic overburden to critical thermal maxima for underyearling Arctic grayling acclimated to 15°C. [ [- [- [ FIG.12.Relationship of concentration of suspended inorganic paydirt to critical thermal maxima for underyear1ing Arctic grayling acclimated to SoC. -87 - I I I !I I "!!! !!!d f ! !!! !I!I!! I I!!!I Note:Points represent mean; bars,95%confidence interval for 10 fish. 50 100 1,000 10,000 100,000 NOMINAL PAYDIRT CONCENTRATION (mg-L-1) I controls I o 28.0 29.0 23.0 :::r:~27.0 wo ~26.0 w 0: ::::>~25.0 0: Wa.. ~24.0w I- -oU- [ [- r~ [ [ [ [ [ [ ...c [ G.·.•.LJ [ [ [ [ [- [- [ -89 - C 280 clntro,s [ [ [' n [ [ [ n l 360 340 320 300 Note:Points represent mean; bars,95%confidence interval for 10 fish. ... 9 9 , E- I 260~wo o 240 I- w ~220 I- 200 180 160 100,000 I I I I I !"I !!!!!"!!!!!!d o ___-.,.,"!II!!!I J 50 100 1,000 10,000 NOMINAL PAYDIRT CONCENTRATION (mg·C l ) FIG.13.Relationship of concentration of inorganic paydirt to time to death in sealed jar bioassays for underyear1ing Arctic grayling acclim- ated to 15°C and tested at 20°C. -91 - Note:Points represent mean; bars,95%confidence interval for 10 fish.2.4 2.5 2.3 -T..J 2.2e --controlsC' /E- z 2.1lLJ (!)>-x 0 2.0 0 lLJ>..J 0 1.9CJ) CJ) 0 ..J 1.8<{ ::> 0 CJ) lLJ 1.7a::: [ [ [, c [ [ c c o c~ .....i 50 100 1,000 10,000 NOMINAL PAYDIRT CONCENTRATION (mgeL-1) 1.6 1.5 I !!I I !I II !!!I!,Ii I I I I!!!I ! !! ! !", 100,000 FIG.14.Relationship of concentration of inorganic paydirt to tolerance to hypoxia in sealed jar bioassays for underyearling Arctic grayling ac- climated to lSoC and tested at 20°C. FIG.15.Relationship of concentration of inorganic paydirt to time to death in sealed jar bioassays for underyearling Arctic grayling acclim- ated to sOe and tested at lOoe. -93 - !I !I !!.I!I I !I I "!I I I I "I Note:Points represent mean; bars,95%confidence interval for 10 fish. 50 100 1,000 10,000 100,000 NOMINAL PAYDIRT CONCENTRATION (mg·C> I I ! !!!!d controls I 360 560 540 380 420 580 400 600 520-c: E-500 J: ~ L1J 4800 0t- L1J 460 ~ t- 440 [ [' [ [ [ [ [ [ c. [ [ E [ [ [ [ l- [. [ FIG.16.Relationship of concentration of inorganic paydirt to tolerance to hypoxia in sealed jar bioassays for underyearling Arctic grayling ac- climated to 5°C and tested at lODe. 50 100 1,000 10,000 NOMINAL PAYDIRT CONCENTRATION (mg-L 1 ) 100,000 I !!,.!!" Note:Points represent mean; bars,95%confidence interval for 10 fish. I I I !II" -95 - !!I I I!,II!I !! !"I controls I 1.00 1.80 1.90 0.80 0.90 2.00 -1.70~•0' E....., z 1.60 lJJ (!) ~ X 1.500 c lJJ ~1.400 (J) (J) c 1.30 ...J«::>c (J)1.20w 0:: 1.10 [ [, [ [ [ [ [ c [ .# [ [ [ [ [ [ [ [- [~ l [ [ [ I' L [ [ [ [ [ .. [ C C [ controls 260 _1_ 240 2 220-c E-I 200 ~w o 180 o ~ ~160 ~ 140 120 -97 - Note:Points represent mean; bars,95%confidence interval for 10 fish. [ [. [- [ 1001-1.,,'1-'.....1....'.'...Io'.'.l.I''u.I_...L......I.'...L.'...I.'...I.''''''''.I.'l.L"_.....I.__,I.o.,I,I...l'...l'I.I'..'.I.I"_.....I._...'...l'-'-'...''..,.'.w'Io501001,000 10,000 100,000 NOMINAL OVERBURDEN CONCENTRATION (mg-["I) (dry weight basis) FIG.17.Relationship of concentration of organic overburden to time to death in sealed jar bioassays for underyearling Arctic grayling acclimated to lSoC and tested at 20°C. FIG.18.Relationship of concentration of organic overburden to tolerance to hypoxia in sealed jar bioassays for underyearling Arctic grayling acclim-o 0atedto15Candtestedat20 C. !!! !!!d 100,000 (mg·L-') ! !!!!I "!! !!!!d Note:Points represent mean; bars,95%confidence interval for 10 fish. 50 100 1,000 10,000 NOMINAL OVERBURDEN CONCENTRATION (dry weight basis) controls _I 1.9 1.6 1.8 1.7 1.5 ..J 2.0 <{ ::>o (f)wa:: • C' E 2.6 -99- 2.7 2.5 2.8 z 2.3w (!) >-X o 2.2 ow ~o 2.1 (f) (f) o ...... -I..J 2.4 .J -101 - 1.7 final control I 1.4 Note:Points represent mean; bars,95%confidence interval for 10 fish. 1.5 ...."""""1')1"'-10,.....,..,j,'..I'..,j,'.I.'.1.1,,_......1..0.-.'......100'.'.1.'.',I"I"...I_......_~,..,j,'.....1..'1-'I.l''.I.'....1_....".....,j'.....'..,j1..,j'o.l'.I.'..'I 0.6 a r 50 100 1,000 10,000 100,000 NOMINAL PAYDIRT CONCENTRATION (mg"C') 1.6 0.7 1.8 0.9 0.8 FIG.19.Relationship of concentration of suspended inorganic paydirt to blood leucocrit values for underyearling Arctic grayling acclimated to ISoC and exposed to sediment for 24 h. '0 1.30"- I- 0:::1.2u 0u ::::>w ..J 1.1 -initial control 1.0 [ ['.-- [. [ [ [ [ [' [ [ [ [ [ [ ..c [ [ [ [ FIG.20.Relationship of concentration of suspended organic overburden to blood leucocrit values for underyearling Arctic grayling acclimated to 15°C and exposed to sediment for 24 h. !!I !!'"II 100,000 (mg-L-1) Note:Points represent mean; bars,95%confidence interval for 10 fish.-final control .1 ..IinitiO control _~II "",,1 "I'!!,1 I 11!!,d 0.50 a i 50 100 1,000 10,000 NOMINAL OVERBURDEN CONCENTRATION (dry weight basis) 0.90 0.80 1.20 1.30 0.60 0.70 lAO 1.50 1.60 1.80 -103 - 1.70 .-1.10 0::uou::::>1.00 UJ --l - "*'- [ [, [ [ [ [ [ [ n .-c [ c [ [ [ [ [. [~ [ FIG.21.Relationship of concentration of suspended inorganic paydirt to blood sugar values for underyearling Arctic grayling acclimated to lSoC and exposed to sediment for 24 h. 100,000 !I ! !!I!I -105 - Note:Points represent mean; bars,95%confidence interval for 10 fish. L..-.L.....II!!! !!!"!!!!,!,I !!f !!!!, 50 0 J 50 100 1,000 10,000 NOMINAL PAYDIRT CONCENTRATION (mg-L-1) 60 140 160 150 [ [ [' [ [ [ [ C 0:.'·.~ ~ C t> [ C [ [ [ [ [. C· [ !!!!!"I 100,000 -I(mg-L ) I I !I I II I -107 - Note:Points represent mean; bars,95%confidence interval for 10 fish. 50 100 1,000 10,000 NOMINAL OVERBURDEN CONCENTRATION (dry weight basis) 50 140 150 FIG.22.Relationship of concentration of suspended organic overburden to blood sugar values for underyearling Arctic grayling acclimated to 15°C and exposed to sediment for 24 h. 130 -a-e 0'120 E- lLJen 1100u:::> ...J c:>100 <{ ~en <{90...Ja.. 80 70 ~-finalcontrol -initial control 60 [ [ [; [ [ [ [ [ D ... [ [ L [ [ [ L L~ [. [ FIG.23.Relationship of concentration of suspended inorganic paydirt to blood sugar values for underyearling Arctic grayling acclimated to 5°C and exposed to sediment for 96 h. 50 100 1,000 10,000 NOMINAL PAYDIRT CONCENTRATION (mg·L-1) 100,000 I I!'I "II I I I!'"I I I I 1,,1 -109 - Note:Points represent mean; bars,95%confidence interval for 10 fish. I !I I I I "J20 220 200 180 -~1600 0' E....., UJ 140C/) 0u :::>-l 120<!> « :!: C/)100« -la. 80 60 f-contro, 40 [ [ -4[ [ [ [ [ [ [ ,.. [ [ c [ [ [ l [' u· [. [ 1100 1000 800 600 ~:::> -l-eu. 400 200 o TURBIDITY (FTU) TOTAL RESIDUE (mg-L-1) ,.-...... •_--~;......_...•It -_-_.,.. ..............""',,... 23301230 05/08/82 II 30 06/081 82 2330 1130 07 1 08 1 82 TIM E (h) 2330 1130 08 1 08 1 82 2330 1130 09 1 08 1 82 2130 HOUR DATE FIG.24.Illustration of concentration of suspended sediment (total residue) and turbidity within cages held in Highet Creek during August 1982. [ ." 2300 2000 1500 1000 ............ --------.TURBIDITY (nUl TOTAL RESIDUE (mg'L-I l ....,. "',,. '.' II I I I I I I I I I I I I I " I ,.'"I I • I ,• I "\I I ."" 1 ',Ii ...· I,' \, I I I I I I I I I I,I,,.._---- .' o 1720 2320 10/09182 1120 11/09/82 2320 1120 12/09/82 2320 1120 13/09/82 TIM E (hl 2320 1120 14/09/82 23 20 1120 1920 15/09/82 HOUR DATE FIG.25.Illustration of concentration of suspended sediment (total residue) and turbidity within cages held in Highet Creek during September 1982. -11 5 - FIG. 26. Gill filaments of underyearling Arctic grayling captured from Minto Creek during September 1982. Note normal appearance of secondary lamellae (a). 300X. ~ c I FIG.27. Gill filaments of underyearling Arctic grayling captured from Minto Creek and held in a cage within Minto Creek for 5 days during September 1982. Note moderate hypertrophy (increase in cell size) and hyperplasia (in- crease in cell numbers) of lamellar epithelium (b), and presence of large numbers of ectoparasites (c). 300X. -117 - APPENDIX 1.Summary of the aquatic biophysical characteristics for the Highet Creek and Minto Creek caged fish sites during August and September 1982. Site [ [ [ v [ p [ [ L~ I' L~ IL~ [ Variable riparian vegetation channel cover biota abundance predominant flow channel channel width (m) mid-channel depth (m) debris abundance bed material Type coniferous deciduous underbrush ground crown overhang aquatic plants stream invertebrates algae August September Minto Creek very few Picea very few dense Salix and Alnus continuous cover of grasses and berries nil low/moderate moderate moderate moderate glide 7.6 0.4 moderate 60%fines,40%gravel 0.69a ,0.43b 0.23d Highet Creek several patches of Picea several (Populus tremuloids, P.trichocarpa, and Betula) continuous cover of Salix and Alnus few patches of grasses and mosses moderate moderate nil low low riffle backwater 3.0 0.5 low 100%fines at surface 0.32c 0.22d [. [ ~DiScharge gauged on August 5,1982. Discharge gauged on August 10,1982. ~DiScharge gauged on August 7,1982. Discharge gauged on September 13,1982. '",r;r-l r-l r-:i r-J r-J r-J c--:J r-TI {~.r-J t\[T"'j L'1 r-J r-J r-l ["'""""l r-7 ~rJ ~I ~l _' ,...----,, I APPENDIX 3.Physical/chemical characteristics during 4-day survival test 0with15 C-acc1imated underyear1ing Arctic grayling exposed to organic overburden suspensions. Time Variable Nominal overburden concentration (h)(mg'L-l) 0 50 100 1 t OOO 5 t OOO 10 t ODO 50 t OOO temperature (OC)15.1 15.0 15.0 15.0 15.1 15.1 15.0 oxygen (mg'L-1 )9.2 9.3 9.2 9.2 9.2 9.3 9.2 0 pH 6.9 6.8 6.9 6.8 6.8 6.8 6.9 conductanced 15 15 15 15 20 25 30 temperature 15.4 15.1 15.3 15.3 15.3 15.2 15.3 oxygen 9.1 9.3 9.2 9.2 9.3 9.1 9.3 24 pH 6.9 7.0 6.9 6.9 6.9 6.8 6.9 conductance 18 18 16 20 25 30 35 I-' I-' 15.2 15.2 15.1 15.2 15.1 15.1 15.2 \0temperature 9.3 9.3 9.2 9.2 9.2 9.3 9.3 If.)oxygen ;/.~. 48 pH 6.9 7.0 6.9 6.0 7.0 6.9 6.9 conductance 20 20 20 20 25 35 40 temperature 15.1 15.3 15.1 15.2 15.1 15.1 15.2 oxygen 9.2 -9.1 9.2 9.2 9.1 9.2 9.2 72 pH 6.8 6.9 6.9 6.9 6.8 6.9 6.9 conductance 20 20 20 20 25 35 35 temperature 15.0 15.1 15.0 15.0 15.1 15.1 15.0 oxygen 9.3 9.2 9.3 9.3 9.2 9.2 9.2 96 pH 6.8 6.8 6.9 -6.9 6.8 6.8 6~9 conductance 20 20 20 25 25 30 35 d llmho'cm-1. APPENDIX 4.Physica1(ch eIlJ.ica.1 cnara.cteri~tic~during 4...day l;lurviva1 test wtth 5 0 C....acclimated underyearlingArctic grayling e:8:l?osedto inorganic paydirtsuspensions. .... Time Variable .Nominalpaydirt concentration (h)(ms'L-1 ) 0 1,000 5,000 10,000 20,000 100,000 temperature (OC)5.0 5.1 5.0 5.0 5.1 5.0 0 oxygen (mg·L-1 )10.8 10.8 10.9 10.7 10.8 10.8 pH 6.8 6.7 6.7 6.7 6.6 6.7 conductancea 18 18 20 30 20 40 temperature 5.2 5.2 5.1 5.2 5.0 5.2 24 oxygen 10.6 10.6 10.5 10.7 10.7 10.6 pH 6.7 6.7 6.8 6.7 6.6 6.7 conductance 18 1~20 30 25 50 ~ N 0temperature4.7 4.4 4.9 4.3 4.6 4.2 48 oxygen 10.8 10.8 10.9 10.7 10.8 10.9 pH 6.8 6.9 6.8 6.8 6.8 6.8 conductance 15 17 18 23 32 35 temperature 5.3 5.0 5.1 5.0 4.9 4.7 72 oxygen 10.9 10.7 10.8 10.8 10.8 10.8 pH 6.7 6.6 6.6 6.7 6.7 6.7 conductance 15 18 22 25 32 37 temperature 5.1 5.0 5.0 5.2 5.3 4.9 96 oxygen 10.8 10.8 10.9 10.9 10.6 .10.8 pH 6.7 6.7 6.5 6.5 6.6 6.6 conductance 18 21 22 29 32 40 a -1llmho·cm • -121 - APPENDIX 5.Residue and turbidity values within a cage held in Highet Creek during --<'the August 1982 in-situ fish survival test. \Date Time Sample Total Total Total Non-Turbidity ~\"(h)noo residue fixed volatile filtrable (FTll) (mg °L -1)residue resi~ye resid~r (mg'L-1)(mgoL )(mgoL ) 05/08/82 1330 EX 1 335 279 56 210 180 1430 EX 2 435 411 24 347 240 1530 EX 3 416 388 28 314 340 1630 EX 4 391 369 22 310 380 1730 EX 5 409 385 24 318 380 [1830 EX 6 381 355 26 29B.360 1930 EX 7 360 339 21 262 320 2030 EX 8 317 304 13 194 280 2130 EX 9 309 290 19 214 260 2230 EX 10 290 277 13 200 225 2330 EX 11 308 285 23 206 250 06/08/82 0030 EX 12 333 310 23 230 295 [0130 EX 13 335 307 28 242 310 0230 EX 14 349 307 42 250 325 fl 0330 EX 15 341 304 37 230 300 P 0430 EX 16 341 313 28 206 290 l 0530 EX 17 350 319 31 262 310 {0630 EX 18 360 333 27 298 310 [0730 EX 19 382 344 38 246 '.300 0830 EX 20 322 291 31 166 280 0930 EX 21 285 265 20 184 190 1030 EX 22 222 198 24 134 130 L 1130 EX 23 208 197 11 104 115 1230 EX 24 365 319 46 244 290 1330 EX 25 466 431 35 392 500 [1430 EX 26 713 668 45 608 950 1530 EX 27 584 543 41 486 700 1630 EX 28 510 482 28 424 700 [1730a EX 29 292 259 33 180 165 1830 EX 30 283 270 13 164 170 1930 EX 31 JJ 2030 EX 32 214 201 13 128 120r2130EX33 L.]2230 EX 34 294 260 34 208 250 2330 EX 35 [07/08/82 0030 EX 36 238 220 18 138 185 0130 EX 37 0230 EX 38 215 203 12 126 150 r--0330 EX 39 0430 EX 40 161 148 13 54 60L'"0530 EX 41 [~ [ -122 -[ r',.~, APPENDIX 5 (cont.)(, (>~~ Sample Total Total Total Non-Turbidity LDateTime (h)no.residue,fixed volatile filtrable (FTU)r(I!lg'.~-1),residue resi~ye residue L(mg.L -1)(mg·L )(mg.L -1) 07/08/82 0630 EX 42 192 '171 21 60 55 [(cont.)0730 EX 43 0830 EX 44 218 187 31 108 65 0930 EX 45 [1030 EX 46 185 173 12 _84 45 1130 EX 47 1230 EX 48 <L23 112 11 16 17 [1330 EX 49 -- 1430 EX 50 96 89 <10 <20 4.8 1530 EX 51 [1630 EX 52 109 86 23 <20 4.6 1730 EX 53 1830 EX 54 118 118 <10 '<20 15 1930 EX 55 35 [2030 EX 56 186 172 14 70 44 2130 EX 57 q 2230 "EX 58 202 195 <10 92 45 r2330'EX 59 U 08/08/82 0030 EX 60 190 195 <10 94 85 '-", 0130 EX 61 [0230 EX 62 208 202 <10 92 80 0330 EX 63 0430 EX 64 168 174 <10 74 65 [0530 EX 65 0630 EX 66 198 183 15 94 54 0730 EX 67 0830 EX 68 157 152 40 38 43 [0930 EX 69 36 1030 EX 70 1130 EX 71 L1230EX72918840<20 6.8 1330 EX 73 1430 EX 74 92 80 12 <20 3.0 [1530 EX 75 1630 EX 76 101 79 22 <20 3.6 1730 EX 77 1830 EX 78 79 79 <10 i.20 3.8 [1930 EX 79 2030 EX 80 105 96 <10 20 20 2130 EX 81 f'2230 EX 82 154 157 <10 50 26 I?L-"2330 EX 83 T' "L [ -123 - -. APPENDIX 5 (cant.) -~~ Date Time Sample Total Total Total Non-Turbidity '(h)noo residue fixed.,volatile filtrable (FTU) (mgOL-1)residue resi~'fe residue (mgoL -1)(mg °L )(mg °L -1) 09/08/82 0030 EX 84 149 159 <10 34 38 0130 EX 85 I'0230 EX 86·184 165 19 38 38 Lo 0330 EX 87 - 0430 EX 88 184 166 18 68 55 0530 EX 89 [0630 EX 90 178 157 21 72 55 0730 EX 91 0830 EX 92 197 176 21 82 55 [0930 EX 93 1030 EX 94 147 136 11 40 53 1130 EX 95 [1230 EX 96 112 105 <10 <20 20 1330 EX 97 1430 EX 98 115 97 18 <20 8 0 0 p 1530 EX 99 [1630 EX 100 117 92 25 <20 8.0 1730 EX 101 I 1830 EX 102 112 90 22 20 5.0 [1930 EX 103 2030 EX 104 111 87 24 <20 7.5 2130 EX 105 [ 2230 EX 106 169 131 38 58 30 2330 EX 107 10/08/82 0030 EX 108 [0130 EX 109 0230 EX 110 199 172 27 88 50 0330 EX 111 0430 EX 112 173 152 21 92 65 [0530 EX 113 0630 EX 114 190 168 22 66 68 0730 EX 115 [0830 EX 116 230 211 19 58 85 0930 EX 117 1030 EX 118 147 125 22 34 25 [1130 EX 119 1230 EX 120 108 77 31 <20 11 1330 EX 121 1430 EX 122 106 89 17 <20 14 1'--1530 EX 123 L<"1630 EX 124 106 97 <10 28 31 [,..,. [ -124 -[ APPENDIX 5 (cant.)[ ~Fish placed in cages. Not analysed ° Date Time Sample Total Total Total Non-Turbidity (h)no.residue fixed volatile filtrable (FTU) (mgOL -1)residue resi~1fe residue (mg °L -1)(mg·L )(mg.L-1) 10/08/82 1730 EX 125 (cant.)1830 EX 126 107 90 17 <20 15 1930 EX 127 2030 EX 128 142 126 16 54 35 2130 EX 129 2230 EX 130 142 130 12 54 30 =======================================================l~ [ [ [ [ [ [ o [ (~ C [ [ [ [ [ [ -125 - -, " APPENDIX 6.Residue and turbidity values within a cage held in Highet Creek duripg -,)the September 1982 in-situ fish survival test. ,j Date Time Sample Total Total Total Non-Turbidity (h)no.residue 'fued volatile filtrable (FTT]) (mg·L-1 )"residue residue residue (mgoL-1 )(mg·L-1 )(mg.L -1) 10/09/82 1530 EX 1 504 471 33 396 350 1630 EX 2 -b 1730a EX 3 429 404 25 308 320 1830 EX 4 1930 EX 5 384 360 24 270 270 2030 EX 6 2130 EX 7 260 241 19 144 150 2230 EX 8 2330 EX 9 333 307 26 236 210 11/09/82 0030 EX 10 ~0130 EX 11 321 295 26 214 220 0230 EX 12",0330 EX 13 329 299 30 226 230 0430 EX 14 0530 EX 15 454 419 35 336 370 9 0630 EX 16 0730 EX 17 492 452 40 326 420 L 0830 EX 18 0930 EX 19 1030 EX 20 1130 EX 21 .;,;.- 1230 EX 22 373 340 33 108 245 1330 EX 23 292 262 30 222 150 1430 EX 24 1530 EX 25 600 556 44 404 410 1630 EX 26 1730 EX 27 599 550 49 484 570 -~1830 EX 28 1930 EX 29 592 545 47 470 510 2030 EX 30 L.J 2130 EX 31 819 754 65 700 1000 2230 EX 32 2330 EX 33 724 671 53 610 820 12/09/82 0030 EX 34 0130 EX 35 629 582 47 490 750 l~"0230 EX 36 0330 EX 37 494 454 40 384 460 0430 EX 38 I'0530 EX 39 688 635 53 558 720 i 0630 EX 40L......,~ 0730 EX 41 492 463 29 358 400 [~0830 EX 42 [ -127 - -129 - APPENDIX 7 (cont.) =',J Date Time Sample Total Total Total Non-Turbidity (h)no.residue fixed volatile filtrable (FTU) (mg ·L:"l)residu~resi~ye resid~r (mg.L-1)(mg·L )(mg·L ) 08/08/82 0820 43 (cont.)0920 44 122 101 21 22 1.3 ,.-,1020 45 1120 46 130 101 29 20 0.8 1220 47 1320 48 139 115 24 20 1.4[1420 49 1520 50 128 106 22 40 1.6 1620 51 [1720 52 126 95 31 <20 1.3 1820 53 1920 54 149 117 32 <20 1.1 [2020 55 - 2120 56 148 115 33 <20 1.1 222.0 57 •2320 58 145 114 31 <20 1.1 [09/08/82 0020 59 r 0120 60 148 116 32 <-20 1.1 0220 61 [0320 62 141 119 22 <20 0.8 0420 63 0520 64 141 121 20 <20 0.7 C 0620 65 0720 66 147 113 34 <20 0.8 0820 67 [0920 68 152 114 38 <20 1.4 1020 69 ~)1120 70 149 101 48 24 1.1 1220 71 [1320 72 146 109 37 24 1.3 1420 73 1520 74 141 106 35 <20 1.3 [1620 75 1720 76 1820 77 L 1920 78 131 95 36 <20 1.3 2020 79 2120 80 135 96 39 <20 1.1 2220 81 [-2320 82 [~. L -130 - APPENDIX 7 (cant.) Date Time Sample Total Tot.a1 Total Non-Turbidity (h)no"residue fixed volatile filtrable (FTU) (mg"L-1)residue resi~'te resid~r (mg"L '-1)(mg"L )(mg"L ) 10/08/82 0020 83 0120 84 133 98 35 <20 1.1 0220 85 0320 86 130 96 34 22 1.1 0420 87 0520 88 137 105 32 24 1.1 0620 89 0720 90 "154 104 50 24 1.6 0820 91 0920 92 153 110 43 <20 1.6 1020 93 1120 94 147 100 47 <20 1.2 1220 95 1320 96 130 100 30 <20 1.2 1420 97 1520 98 144 114 30 <20 1.2 1620 99 1720 100 141 113 28 <20 1.8 :~Fish placed in cages. Not analysed" [ t [ [ [ [ C [ t "(\"t [ [ [ [ [ L J~ L [ .,...132 -[ l' APPENDIX 8 (cant.)L l (-, Date Time Sample Non-Turbidity L. (h)no.filtrable (FTU) residue [(mgoL-1 ) 12/09/82 1230 43 6 0.8 [(cant.)1330 44 1430 45 14 0.9 1530 46 [1630 47 0.9 1730 48 <5 1830 49 0.9 [1930 50 5 2030 51 16 1.5 2130 52 2230 53 <5 0.8 C23305412 13/09/82 0030 55 1.5 0130 56 C0230570.6 0330 58 0430 59 1.2 t0530608 0630 61 to- 0730 62 0.6 [108306380.8 0930 64 <0.5 Jj 1030 65 1130 66 <5 [1230 67 10 10 " 1330 68 1430 69 <5 0.9 [1530 70 1630 71 <5 1.0 1730 72 D183073<5 1.0 1930 74 <5 2030 75 0.7 2130 76 n 2230 77 <5 0.8 U 2330 78 14/09/82 0030 79 <5 0.8 U0130806 0230 81 0.6 0330 82 fi043083e-:.-J 0530 84 (1 :td"'~ U U r,>I . l~'- r l, [ [ [ [ [ c l [ [ [ r-' L r L [ -133 "- APPENDIX 8 (cant.) .Date Time Sample.Non-Turbidity (h)no.filtrable (FTU) residue (mgoL-1) . 14/09/82 0630 85 (cant.)0730 86 <5 0.7 0830 87 0930 88 1030 89 1130 90 0.9 1230 91 0.9 1330 92 5 1430 93 <5 1.2 1530 94 1630 95 <5 1.2 1730 96 1830 97 <5 1.1 1930 98 <5 2030 99 1.1 2130 100 <5 0.8 2230 101 2330 102 8 15/09/82 0030 103 0.7 0130 104 0230 105 11 1.2 0330 106 0430 107 0.5 0530 108 0630 109 6 0.5 0730 110 0830 III 12 0.9 0930 112 <5 1030 113 <0.5 1130 114 1230 115 <5 <0.5 ~Fish placed in cages. Not analysed. -134 - APPENDIX 9.Comparison of suspended sediment and turbidity values for triplicate water samples taken from within or outside of a Highet Creek cage during the August and September 1982 in-situ fish survival tests. Date Sample Nonfiltrable Total Turbidity no.residue residue (FTU) (mgoL-l)(mgoL-l) 06/08/82 EX 28 a b 424 482 700 28-2 364 436 580 28-3b 340 456 600 06/08/82 EX 30 a 164 270 170 30-2b 424 519 300 30-3b 354 455 280 07/08/82 EX 46 a b 84 173 45 46-2 147 316 45 46-3b 147 254 70 07/08/82 EX 55 a b 88 176 35 55-2 98 187 42 55-3b 58 158 39 08/08/82 EX 69 a 34 122 36 69-2b 244 272 50 69-3b 216 375 70 11/09/82 EX 22 a 108 340 245 22-2b 264 380 250 22-3b 314 438 225 13/09/82 EX 79 a b 304 1160 1550 79-2 586 1280 1550 79-3b 514 1160 1550 abSample collected by Isco automatic pump sampler from within cage. Sample collected manually just outside of the cage,at the time that sample "a"was taken. n D n o D [ [ InC,J c [ [ [ [ [ [