V'Hfi 'E‘ELANSJ’ER 0F RADECPHOSQHORUS WEFHN A S'!‘§'.EAM AND ”’5 RELATIORSHIP TO .LEGH? RNWNSITY ANS ENSEC? MIGRATION Thesis {or the: Degree 05" M S. mam/w STATE UiffiEVERSETY Thamas A. ‘Woéfi‘afik 1963 THESB LIBRARY Michigan State University .....l. -rllill' .'1 IO?» ABSTRACT THE TRANSFER OF RADIOPHOSPHORUS WITHIN A STREAM AND VITS RELATIONSHIP TO LIGHT INTENSITY AND INSECT MIGRATION by Thomas A. Wojtalik The present study was conducted to determine the agencies of transfer of phosphorus in a stream ecosystem. On July 10, 1962 twenty- three millicuries of P32 were added to the West Branch of the Sturgeon River. Samples of the water mass were fractionated and eight fractions were analyzed for amounts of radioaCtivity present in the water passing given points in the study area. By comparison of the amounts of tracer present in particular fractions at the different points, loss from the water mass, uptake by the different fractions, change in mode of trans- fer and possible agents of that change were calculated. In earlier work on this stream, stream organisms had been investigated with refer- ence to their specific concentration of stable and radioactive phos- phorus, but not with reference to the possible modes of transfer of the element other than whole organism ingestion by other organisms. Trans- fer modes were investigated indirectly by analyzing the water fractions for soluble organic materials and insoluble organic materials. A study of the movement of aquatic insects showed that an appre- ciable amount of radiophosphorus moved upstream through the migration of both immature forms and the flights of adult insects. Thomas A. Wojtalik Light measuring and integrating instruments were developed and the relationships of light energy reaching the stream to production of periphyton and uptake of P32 in a light area, a dark area, and an inter- mittently lighted area was studied. THE TRANSFER OF RADIOPHOSPHORUS WITHIN A STREAM AND ITS RELATIONSHIP TO LIGHT INTENSITY AND INSECT MIGRATION BY Thomas A. Wojtalik A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Fisheries and Wildlife 1963 ACKNOWLEDGEMENTS During the preparation of this thesis and the gathering of data for it, many people have aided and encouraged the author. To these people, the author would like to express his deep appreciation. The author is particularly indebted to Dr. Robert C. Ball for his constant encouragement and enthusiasm as well as his timely advice on many as- pects of the research conducted, and to Dr. Frank F. Hooper whose many ideas and words of encouragement have proved valuable throughout the study period and during the authors undergraduate work. In the field, the author was assisted by Jack Bails, Eugene Buck, and Jerry Hamelink. For their assistance and help during the field work and while working up the data the author is deeply grateful. To Mr. Clint Harris who designed our light meters and assisted in their placement after calibration, the author gives special thanks. Dr. W. Carl Latta and his staff at the Pigeon River Trout Research Station also contributed a great deal to the project and author by their help and advice. The author also wishes to thank Dr. Phillip J. Clark for his assistance with the statistical analyses. Two fellow graduates who contributed their help and ideas were Michael E. Bender and James Bacon, Jr. who did much of the separation work in 1962 and assisted the author in his studies of migration during the 1961 study, the author is deeply grateful to both. The study was made possible through a grant held by Dr. R. C. Ball ii and Dr. F. F. Hooper from the Atomic Energy Commission [AT (ll-l)655]. The author was supported by a Graduate Research Fellowship from the Institute for Fisheries Research of the Michigan Department of Conservation. iii TABLE OF CONTENTS Page INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Introduction to the Stream and Study Area . . . . . . . . . 5 Sampling Stations . . . . . . . . . . . . . . . . . . . . . 7 Station 3 . . . . . . . . . . . . . . . . . . . . . . . . 8 Station A--Shaded Area . . . . . . . . . . . . . . . . . 8 Station 5 . . . . . . . . . . . . . . . . . . . . . . . . 11 Station B--Light Area . . . . . . . . . . . . . . . . . . 11 Station 8 . . . . . . . . . . . . . . . . . . . . . . . . 11 Station 12 . . . . . . . . . . . . . . . . . . . . . . . 12 Station 14 . . . . . . . . . . . . . . . . . . . . . . . 12 100 Yards Above . . . . . . . . . . . . . . . . . . . . . 12 200 Yards Above . . . . . . . . . . . . . . . . . . . . . 13 300 Yards Above . . . . . . . . . . . . . . . . . . . . . 13 400 Yards Above . . . . . . . . . . . . . . . . . . . . . 13 500 Yards Above . . . . . . . . . . . . . . . . . . . . . 13 METHODS AND PROCEDURES . . . . . . . . . . . . . . . . . . . . . 14 General Methods . . . . . . . . . . . . . . . . . . . . . . 14 Special Considerations with the Isotope . . . . . . . . . 15 Combined Study Methods . . . . . . . . . . . . . . . . . 16 The Isotope . . . . . . . . . . . . . . . . . . . . . . . . l7 IsotOpe Introduction . . . . . . . . . . . . . . . . . . . . 17 iv Page Measurement of Radioactivity . . . . . . . . . . . . . . . . 20 Calculation of Activity . . . . . . . . . . . . . . . . . . 21 Correction Factors . . . . . . . . . . . . . . . . . . . . . 21 BIOLOGICAL SAMPLING PROCEDURES . . . . . . . . . . . . . . . . . 24 Water Samples . . . . . . . . . . . . . . . . . . . . . . . 24 Water Sampling . . . . . . . . . . . . . . . . . . . . . 24 Water Processing . . . . . . . . . . . . . . . . . . . . 25 Transect Collections . . . . . . . . . . . . . . . . . . 29 Processing for Activity . . . . . . . . . . . . . . . . . 3O Mud Samples . . . . . . . . . . . . . . . . . . . . . . . . 31 Mud Sampling . . . . . . . . . . . . . . . . . . . . . . 31 Processing for Activity . . . . . . . . . . . . . . . . . 32 Periphyton Samples . . . . . . . . . . . . . . . . . . . . . 32 Periphyton Sampling . . . . . . . . . . . . . . . . . . . 33 Processing for Activity . . . . . . . . . . . . . . . . . 34 Stream Macrophytes . . . . . . . . . . . . . . . . . . . . . 35 Plant Collections . . . . . . . . . . . . . . . . . . . . 38 Processing for Activity . . . . . . . . . . . . . . . . . 38 Plant Standing Crop and Biomass . . . . . . . . . . . . . 39 Invertebrate Samples . . . . . . . . . . . . . . . . . . . . 40 Invertebrate Collections . . . . . . . . . . . . . . . . 41 Migration Collections . . . . . . . . . . . . . . . . . . 42 Adult Insect Collections . . . . . . . . . . . . . . . . 42 Larval Insect Collections . . . . . . . . . . . . . . . . 46 Drift Collections . . . . . . . . . . . . . . . . . . . . 46 Processing for Activity . . . . . . . . . . . . . . . . . 46 Page Larval Forms . . . . . . . . . . . . . . . . . . . . . 46 Adult Forms . . . . . . . . . . . . . . . . . . . . . 47 Biomass Collections . . . . . . . . . . . . . . . . . . . 47 Biomass Enumeration and Weights . . . . . . . . . . . . . 48 Fish Samples . . . . . . . . . . . . . . . . . . . . . . . . 48 Collection . . . . . . . . . . . . . . . . . . . . . . . 48 Processing for Activity . . . . . . . . . . . . . . . . . 49 Fish Biomass . . . . . . . . . . . . . . . . . . . . . . 49 RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . . . . . 51 Water Samples . . . . . . . . . . . . . . . . . . . . . . . 51 Sediment Samples . . . . . . . . . . . . . . . . . . . . . . 70 Periphyton or Aufwuchs . . . . . . . . . . . . . . . . . . . 80 Macrophytes . . . . . . . . . . . . . . . . . . . . . . . . 104 Macrophyte Biomass and Production . . . . . . . . . . . . . 138 Invertebrates . . . . . . . . . . . . . . . . . . . . . . . 141 Migration . . . . . . . . . . . . . . . . . . . . . . . . . 163 Fish Samples . . . . . . . . . . . . . . . . . . . . . . . . 175 Fish Biomass and Concentration of P32 . . . . . . . . . . . 186 Light-Dark Study . . . . . . . . . . . . . . . . . . . . . . 190 Light Measurement . . . . . . . . . . . . . . . . . . . . 193 Calibration . . . . . . . . . . . . . . . . . . . . . . . 194 Area Comparisons . . . . . . . . . . . . . . . . . . . . 199 SUMMARY . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203 LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . 205 vi Table 2a. 2b. 10. ll. 12. 13. LIST OF TABLES Radioactivity levels of the water fractions at Stations 3, 8, and 14 in corrected micromicrocuries per milliliter Concentration of isotope, amount retained, amount leaving each station Total millicuries retained and leaving each station . Radioactivity levels in eight fractions of water taken as P32 spike passed Stations 3, 8, and 14 . Concentrations of radiophOSphorus in diatoms and bacteria Comparison of uptake by water fractions in different years at the different stations Stations 8 and 12 sediment activity in sand and mud fractions, their losses, and gains Sediment fractions uptake by area and rate Drift diatom composition by station, by week, between July 9, 1962 and August 20, 1962 Summer composition of drift diatoms and mean weekly composition during the first six weeks of the study . Periphyton radioactivity in corrected counts per minute per gram of wet material Radioactivity levels of plant material from the West Branch of the Sturgeon River collected in 1962 . Variance and standard deviation of plant activity levels per species Macrophyte composition and production of three major Species in 1962 . vii Page 54 56 56 59 66 71 77 79 82 83 90 136 137 139 Table 14. 15. 16. 17. l8. 19. 20. Macrophyte production figures per unit area and phosphorus pool contained in each species Insect radioactivity levels in micromicrocuries per gram of wet insect material at weekly intervals Radioactivity levels of adult insects collected by ultra-violet light traps Radioactivity levels in immature insects by date and distance upstream above the point of isotope introduction in 1961 Radioactivity levels in immature insects by date and distance upstream above the point of isotope introduction in 1962 Trout population estimates of West Branch and their concentration of isotope. . . . . . . . . . . Data on light reaching stream . . . . . . . . . . . viii Page 140 144 165 168 170 187 198 Figure 10. ll. 12. 13. 14. 15. LIST OF FIGURES Map of the West Branch of the Sturgeon River with sampling stations and stream study areas designated . Stage readings of the stream gauge by date A.Ma1aise directional trap Total activity passage past Stations 3, 8, and 14 Illustration of the tracer accumulation among the sediments at Stations 8 and 12 Mean number of diatoms of nine major genera by weeks Mean number of drift diatoms by station for total summer and mean radioactivity of periphyton by station . Activity levels of periphyton at Stations 3, 5, and 8 . Activity levels of periphyton at Stations 12 and 14 Activity curves of substrates spiked with radioactive phosphorus at Stations 3 and 5 respectively, and then moved to Allan Creek for the remainder of the study . Periphyton radioactivity levels and corresponding water levels Comparative Station 8 uptake and loss for Chara sp. Comparative Station 8 uptake and loss for Fontinalis antipyretica Comparative Station 8 uptake and loss for Potamogeton sp. Levels of standing crop of macrophytes as related to fertilization . ix Page 18 43 57 73 85 87 91 93 97 100 106 108 110 114 Figure 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. Stations levels Stations levels. Stations Stations Stations Stations Stations Stations Stations levels Stations levels Stations Simulium Stations 3, Sp. 3. Ephemerella 5, and and 14 5, and and 14 5, and and 14 5, and and 14 5, and and 14 Chara sp. activity levels 8, and 14 8, and 14 needhami. Stations 3, Ephemerella 8, and 14 cornuta . Stations 3. 8, and 14 Atherix variegata . Stations 3, 8, and 14 activity fishfly larvae, Chauloides Sp. activity activity activity activity Stations 3, 8, and 14 activity snail, Physa sp. Stations 3, 8, and 14 activity stonefly, Pteronarcys Sp. Stations 3, 8, and 14 activity Hexagenia Sp. levels levels levels levels levels levels levels levels 8 Fontinalis antipyretica activity Fontinalis antipyretica activity 8 Potamogeton sp. activity levels Potamogeton sp. activity levels 8 Chara sp. activity levels 8 Ranunculus sp. activity levels Ranunculus Sp. activity levels 8 Nasturium officinale activity Nasturium officinale activity of the blackfly, of of of of of of of the the the 32 the the the mayfly, mayfly, snipefly, in the pouch riffle mayfly, Curve of individual variability within a particular species at a given station demonstrated with muddlers at Stations 8 and 12 Page 116 118 120 122 124 126 128 130 132 134 146 148 150 152 154 156 158 160 176 Figure Page 35. Regressions of activity with time in days . . . . . . . . 179 36. Regression of activity with time in days . . . . . . . . 181 37. Regression of activity of fish by time in days . . . . . 183 38. A light sensing unit in its immersible covering, with glass hemisphere and cord to shore counter and power source . . . . . . . . . . . . . . . . . . . . . . . . . 196 xi . u. INTRODUCTION In many aquatic ecosystems phosphorus has been found to be a consistently limiting factor to the productivity. Its limited avail- ability was evident at all trophic levels in the West Branch of the Sturgeon River and it appeared to be especially limiting at the primary level since all phOSphorus fertilization experiments indicated a rapid uptake and subsequent bloom of periphyton near the source of fertilizer introduction which then gradually decreased with time. In the fertilization studies conducted on the West Branch of the Sturgeon River a series of phosphorus additions were conducted over a period of more than five years. These additions were at its source, Hoffman Lake; at the lake outlet, and at several points approximately five to eight miles downstream. Each study indicated phosphorus uptake was rapid in the immediate area of introduction, but little uptake in any of the trophic levels could be found in downstream areas. There- fore in 1958, additions of tracer phosphorus were made using the radio- active material P32. It was intended that pathways of phosphorus transfer be followed from the inorganic form introduced, through the primary producers to the primary consumers, secondary consumers and all other levels up to and including fish which were considered the terminus for the ecosystem. A secondary consideration of the fish terminus was from a public health view of contamination due to fallout over watersheds which furnish both food and water to humans. 1 2 The West Branch of the Sturgeon River is a representative of the small, cold-water trout streams which formerly were typical of Michigan. Its source is a marl lake, but most of its summer flow of approximately 49.7 cubic feet per second at nine miles from its source, is supplied by ground water fed out of underground springs present in the surrounding forest covered moraines. Its morphology is one of nearly equal proportions of riffles and pools or runs which are under- lain by a marly gravel interspersed with larger marl concretions. The average width is 28.3 feet and the average depth is 1.4 feet for the section used during the present study. From its source to the lower end of the study area, a section approximately nine miles long, it winds through heavy cedar swamps and densely forested valleys. Its summer steady state of phOSphorus concentration was approxi- mately 7 p.p.b. with spring peaks of up to 20 p.p.b. (Correll, 1958) during the high runoff period. Oxygen content was always high due to the large amount of oxygenation by the large nhmber of riffles which keep the stream at least in equilibrium with the atmosphere. Alkalinity was exclusively of the methyl orange variety and maintained itself at 180 p.p.m. Turbidity and color were low during most of the three months of the study, only immediately after heavy rains did the turbidity in- crease appreciably. Large amounts of organic detritus were moved down- stream by continuous saltation and sliding of the fragments over the bottom substrates. In several of the years of the fertilization study the pulse of periphyton close to the source of introduction indicated a rapid uptake of phOSphorus which created a large exchange pool in that area alone. Therefore in the radioactivity studies of 1958, 1959, and 1960 3 pre-tracer additions of two to three times the normal phosphorus load were made to increase the exchange pools. During the 1961 and present Study no pre-tracer additions of any fertilizer were made. Each of the additions of tracer was used to further our knowledge of phosphorus pathways in a lotic ecosystem, trophic levels were indicated (of which some had long been established by other means of study and which were only confirmed with the tracer study), new ones were established and food webs were constructed. Recycling was indicated as was a reserve phosphorus pool in the primary producers (periphyton in particular) which maintained ratios of nitrogen to phosphorus as high as 10:1 when it was commonly believed this ratio was 5:1 (Correll, 1958). Diatoms were believed to be the major primary producers during the early studies because filter feeders like Simulium sp. were always found to be filled with them. The bacterial contribution to a food chain was hard to evaluate and misleading when only stomach analyses were run. Therefore to definitely isolate the bacterial contribution of phosphorus to filter feeders and other parts of the food chain or web the 1961 study -(Bender, 1962) was conducted using Escherichia coli 0111 as the organic bacterial carrier of P32. The results were negative and helped confirm a definite diatom base for the food web of the stream and the pyramid of numbers built thereupon. The 1961 study also raised a question con- cerning the food of the fishfly larvae, Chauloides sp., since they accumulated from 10 to 100 times the activity recorded in earlier studies when the tracer introductions were of an inorganic nature. Their head capsules and mouthparts are definitely not suited to filter, scrape, or catch bacteria. Each of the several inorganic introductions of P32 and the single 4 incorporated organic tracer addition had told much about the food chains and stream food web, but none defined exact proportions con- tributed by different agents that were probably present in the water. Nor did they indicate modes or amounts of change between sampling areas or with increased distance for organic fractions. Therefore, the present study was initiated to do much of this type of isolation of fractions. Water was fractionated into diatom; bacteria; adsorbed; soluble organic; non-water soluble, non-ether soluble; non-water, non- ether, non-acid soluble organic; and small "particulate" fractions. Amounts of P32 were determined in terms of the radioactivity passing a given point while in a given fraction. Changes were followed as well as sources of possible contributions to the total organic fractions, such as plankton contributions and periphyton, plants, insects and fish. Modes of uptake among the sediments were investigated as were areas of deposition of the organic materials and carriers. Movement of the isotope due to insect activity was investigated using drift organisms and actively migrating adult and immature insects in areas above the point of isotope introduction. Movements due to fish activ- ity as well as their possible physiological activity were investigated from the same areas above the point of tracer addition. A secondary consideration concerned the pools of exchangeable phosphorus present and the amount by which they could be altered by fertilization methods especially designed for a given stream. Dry weight and phosphorus conversion factors were determined to milligrams per individual in previous years, especially by Zettelmaier (1961). Productivity figures were evaluated for a lighted, open area of stream and a densely shaded tunnel using submersed light meter units 5 in both areas of similar depth and nearly similar current. The light- dark study was conducted with respect to possible use as a management tool along non-productive, shaded streams. Introduction to the Stream and Study Area The West Branch of the Sturgeon River originates from Hoffman Lake, a 128 acre marl lake located in Charlevoix County, Michigan. From its origin, the general pattern of flow is in and out among wooded moraines that create a northeasterly directed valley. Its entire length from source to confluence with the Sturgeon River south of Wolverine, Michigan is 14 miles. This study was conducted upon a 3230 yard section approximately five miles above its terminus. During the course of its flow the West Branch of the Sturgeon River passes through Cheboygan, Otsego, and Charlevoix Counties, each of which is heavily wooded, glacial country where the river flows. The vegetation about the stream is chiefly of birch, aSpen, alder, and balsam fir on the higher moraines. At streamside it is chiefly cedar, tamarack, aspen, alder, and ninebark. The river is one of the few cold-water trout streams left within the state, its temperature varies between 520 F. and 580 F. during the summer low water months (Clifford, 1959). It remains open throughout the winter and has a flow little increased from that of the summer lows. The majority of summer flow is from underground water supplied from surrounding pockets and springs part way up the side of gravelly moraines. The actual study area was located in T. 33 N, R. 3 W. Its average width was 28.3 feet and its average depth was 1.4 feet. The gradient 6 was sufficient for a morphology of half riffles and half pools or runs. The bottom varies in composition from sand and a marly gravel to silt and detritus. The vast majority was of marly gravel and large marl concretions. General stream conditions were as follows: the stream flow had a mean flow of 43.75 cubic feet per second (Knight, 1961), it entered the area at 38.75 cubic feet per second and left Station 14 at 49.37 cubic feet per second. The gain in flow was from two small tributary creeks and 14 small springs along with underwater contributions of ground water. Rains rarely affect the summer level, its color or turbidity. The alkalinity was normally 180 p.p.m. of methyl orange, and carbonates precipitate in concretion rings of about 3/8 of an inch per year. The total phosphorus concentration during the summer months was approximately 7 parts per billion. A pH of approximately 7.9-8.3 was present during the summer. A high dissolved oxygen content existed due to the riffles and excellent diffusion. Aquatic organisms were abundant, especially aerophilic groups. The chief aquatic vegetation present was in dense beds, of such species as Qhaga sp.; the water moss, Fontinalis antipyretica; and water cress, Nasturium officinale; with scattered clumps of Ranunculus sp.; and a Potamogeton Sp. The aquatic plants were abundant in most of the stream, an exception was a stretch from 300 yards to 600 yards below the point of isotope introduction. It was an area of nearly com- plete canOpy which passed less than 500 foot-candles of sunlight during the day except for scattered light flecks. The trophic structure was topped by three salmonids: the brown trout, Salmo trutta fario; the rainbow, Salmo gairdnerii; and the brook 7 trout, Salvelinus fontinalis in their respective order of abundance. An end in itself as well as a part of the structure, was the sculpin group composed of two members: the eastern slimy sculpin, Cottus cognathus; and the northern mottled sculpin,Cottus bairdii. Most of the typical trout stream insects were present, i.e. members of the Diptera, Ephemeroptera, Megaloptera, Neuroptera, Plecoptera, and Tricoptera. Species represented were ones common to clean, cold water that was swift and well oxygenated. Coleoptera, Hemiptera, and Odonata were not represented in large numbers. Oligochaetes and Hydracarina were also not abundant. Gammarus was represented only at the lowermost station (14), and from there down- stream. No crayfish have ever been found by the author or his associates. A table of area per station, flow, and depth follows: Station Total Area Average Depth Flow 3 17430 ft.2 12.8 ins. 38.74 cfs. 5 21000 ft.2 13.5 ins. 40.15 cfs. 8 37924 ft.2 17.2 ins. 43.50 cfs. 12 134245 ft.2 14.3 ins. 45.56 cfs. 14 _ 52992 ft.2 13.3 ins. 49.75 cfs. Sampling Stations Typical trout streams provide a variety of habitats that must be sampled in their entirety to give a valid representation to an ecosystem nutrient study. In other years, 16 stations had been selected with reference to flow, bottom type, vegetation, and shade. Of these, five were used in 1962 along with six new ones established for particular 8 phases of study. The old stations used were 3, 5, 8, 12, and 14. The new stations were five for migration studies, at 100 yard intervals be- ginning at the point of isotope introduction and proceeding upstream, and a new shade area 130 yards below Station 3 and its opposite light area 200 yards above Station 8 (Figure 1). Samples of water, periphyton, plants, insects and fish were taken at Stations 3, 5, 8, 12, and 14. Collections of insects and fish ‘were made at 100, 200, 300, 400, and 500 yards above the transect of isotope introduction. Light readings and periphyton substrates were collected in both the shaded area and the light area. Area descriptions of the sampling stations are as follows: Station 3 A site located 240 yards downstream from the point of isotope introduction. The immediate sides were well shaded, while the upstream section down to the sampling point was open to the sun. All the section was log strewn. The logs were covered with mats of Fontinalis which grew out of the crevices and marl concretions on the logs. Scattered areas of plants consisted mostly of Chara sp. were present, as were small amounts of Ranunculus sp. and a little greater amount of Nasturium officinale and Potamogeton Sp. Flow was about 38.74 cfs and the average depth was 12.8 inches (Zettelmaier, 1961). The shallow, rapid flow helped concentrate the invertebrates in backwaters and on projecting log surfaces depending upon their requirements. Station A--Shaded Area A 200 foot section of complete or nearly complete canopy over a bare marl gravel with no aquatic vegetation other than periphyton. Its Fig. l.--Map of the West Branch of the Sturgeon River with sampling stations and stream study areas designated. .. 1,0- 1 a / .fl%. 1 .. i wZ_mm>n_03 _ a. wJ:2 Exam 3358 To 59:22 $2500 $9635 .2, my”. .2 mm H HommHn HmUOu N HE\053.M Houou N HB\od:.m Hmuou N Ha\u:= N l cowuumum ea coaueum m coaumum m coaumum .hmooo mam masouwxomn you mouuouuoo .uouHHAHHHB Hod mdapddOhUaa nouowe omuoouuoo a« «H can .w .m coaumum um macauomuw nouns mnu mo mHo>oH hua>auouowumM::.H HAMmoa N w”\oss HB\o=: cowucmuom N moawmuom . ~E\u== o>aumfiaazo .wwwwq vowwmwwm m>aumasaso Hmuoa N waa>mon N .ocoo aowumum .GOHumuw 30G” mgfixrfimd Una.“ UmfiHQH—OH mmHHDUfiHHHE HGUOHII.n—N HEM. .uoUHHHAHHB you moauaoouowsouoaa vouoouuoo .cowumum some wcw>mo~ uaaoam .vocamuou ucsoam .odouomw mo :oHumuucoocoU-.mu mamm~ 0u0>wuumofiwmm-.m 0400uomOvam :00umum .08 00009 aowumuuaoocou IOHOHZ HNuOH Umufl—JOU .EmHGmwuo you 0000500000800008 vmuumuuoo .000muomn can maoumwv 60 wnuonmmonm00vmu mo ma00umuucmocoo-n.q uAm0uo< 009000024 .0800: .Ha\o= .060 .v0 :0 aowuuom HmuoH mo newuuomoum .v0 .60 use meuaD mxmuab 00009 $004 .umuHHHHHHE 00m 00003000008 vmuuouuoo .0C000000 ucoummwwv 650 as 00060 ucmnmwmwv :0 mGOHuomum 000w: mp oxmums mo acmwquEoo-u.m mqm0000 00050000 N0 000 0 00000000-u.0 m00<8 78 Holden, (1961) found an exponential decrease of tracer phosphorus in lake water, which he attributed to sediment incorporation. He has suggested that the loss or rate of removal was proportional to the con- centration present. From his data he developed the formula: where: K = the rate constant c = concentration t = time d = differential interval where the rate constant K can be calculated as the percentage loss in concentration per day. ‘This only applies to the case where the phos- phate was removed by a small unit area of the surface, the surface being the surface of the bottom deposits. If the lentic time factor were changed to distance in a lotic environment the values obtained by the formula may also serve to describe the uptake of available radio- activity. The few measurements available from this study only suggest this possibility (Table 7). Holden further discovered that when 700 micrograms of phosphorus per square centimeter were applied, 85 per cent of the phosphate was taken up by the aerobic zone of the sediments, and only six per cent could be extracted with deci-normal hydrochloric acid. His suggestion was that most of the added phosphorus had been converted to a stable organic form. He further suggests that much of the removed phOSphorus may be converted to forms that were unavailable for later release. Although he did not separate his sediments into graded fractions the 79 .00000000 .000 0 .0G HU .000000000000 .000000000 0000000000 000 00 0000>oo 0000 00005000m u 000< 00009 N0 .000 0000 000 000 000000000 00000000560 000 0003000 0000 0000B n 000< 0000H0 00000. 00000. 00.0 0:: 00 00000. 00000. 00.0 000.000 0000 00 0000.0 0000.0 00000. 00000. 000.0 000.00 00.0 002 0 0000.0 0000.0 00000. 00000. 000.00 000.00 00.0 000.00 0000 0 .004 .UGH .mfi< .UGH .035 .053 Amumfiwummv .um .60 N00000: :0 00000000 .00E\Nc0\oss 0.00< 0.000 0000.00 0000< :000000h 0000000 0003000 000000000: 0000 00000: 000< 00009 N 00009 .00000000 0003000 0000 000 000 000000030 00 0000 0000B .0000 000 0000 00 00000: 000000000 u:050000--.0 00000 80 results of the lake studies were nearly comparable to the results of this study. Both suggest that once the material was incorporated from 'the dissolved inorganic state it was not readily released in an in— organic form. It was also indicated that in both experiments the re- leased adsorbed material was bound to an organic compound. In dis- agreement, the two experiments do not indicate the fact that aerobic sediments take up activity only at a slow rate. Our study indicates an uptake rate of available radioactivity greater than twice that of either the plankton bacteria or diatoms, and more than their combined radioactivity. Periphyton or Aufwuchs Within the West Branch of the Sturgeon River periphyton, com- posed primarily of diatoms, was the major primary producing unit. Every part of the surface exposed to the water and to light was covered. Because of its nearly universal coverage, and the indications from stomach analyses that it was the base of the food chains in the eco- system, periodic samples were taken to determine the amount of peri- phyton and its incorporated P32 accruing on plexiglass substrates. The composition had been examined in previous years, particularly by Clifford (1959) who found that Synedra ulna was the overwhelmingly dominant form in two separate studies one year apart. szbella sp.; Navicula sp.; Cocconeis sp.; and Gomphonema sp.; were other principal diatoms of the plexiglass substrates (Clifford, 1959). The time of sampling and the sampling interval were important to the colonization of the artificial substrates as indicated by Butcher (1932), Young ,(1954) and Blum (1957). Peters (1959) reported a marked seasonal 81 periodicity of algal organisms which attached to artificial substrates in the Red Cedar River, Lansing, Michigan. Of particular interest was the report of Butcher (1932) who could not observe any differences in sessile algae on artificial substrates (glass slides) from that found on natural substrates in the river. Periodicity of species on substrates was not observed during the present study. Diatom chains broke off in the current and dead diatoms were suspended for some distance before settling out. These were sampled quantitatively at all stations at weekly intervals. Table 8 indicates the types collected, relative abundance, and any possible differences in composition at each sampling site. Using a plankton net of No. 25 silk bolting cloth and sampling for two minutes at each station an average of 16.43 diatoms per milli- liter were collected in the drift. These diatoms were of nine major genera. A two-way analysis of variance indicated no significant dif- ference between stations at the 95 per cent level of confidence. By comparison of the types present in 1958 and 1959 with those of the present study a change in composition was evident. Navicula sp. had replaced Synedra ulna as the overwhelming dominant form and two other genera; Fragilaria sp. and Cocconeis sp., respectively were more abun- dant than Synedra in number of occurrences by frequency analysis. Table 9 represents the weekly frequency distributions and composite summer frequency distribution. From Table 9 it can be seen that certain genera may have been changed in numbers present, but with the exception of szbella sp., and Diatoma sp. the other seven genera show no change, in either percentage composition or total numbers, which would be in- dicated by a higher frequency. This would signify cyclic changes of 82 TABLE 8.--Drift diatom composition by station by week, between Based on per cent July 9, 1962 and August 20, 1962. frequency of occurrence of the total composition. Per Cent of Total Composition . July July July July 30 Aug. Aug. weEk Of' 9-16 16-23 23-30 Aug. 6 6-13 13—2 Station 3 Synedra 12.5 2.63 5.71 17.65 7.69 3.85 Navicula 12.5 36.84 45.71 32.35 30.77 23.08 Fragellaria 62.5 68.18 11.43 0 15.38 69.23 Cymbella 6.25 2.63 2.86 O 7.69 0 Tabellaria 0 0 5.71 O 0 O Diatoma O O 2.86 11.76 23.08 3.85 Cocconeis 6.25 2.63 14.29 23.53 15.38 0 Cocinodiscus O O 8.57 5.88 0 O Surrirella 0 0 2.86 5.88 O 0 Station 8 Synedra 17.86 22.22 7.32 4.76 16.67 0 Navicula 64.29 66.67 48.78 52.38 83.33 68.75 Fragellaria 3.57 0 12.20 4.76 O 0 Cymbella 3.57 11.11 0 0 0 O Tabellaria 0 O 14.63 9.52 0 0 Diatoma 0 O 2.44 14.29 0 O Cocconeis 7.14 O 12.20 0 0 12.50 Cocinodiscus O O 0 4.76 0 12.50 Surrirella O 0 2.44 4.76 0 6.25 Station 14 Synedra 0 0 8.33 4.26 O 10.34 Navicula 44.44 16.67 38.33 42.55 0 24.14 Fragellaria 33.33 33.33 30.00 36.17 0 O Cymbella 11.11 16.67 6.67 6.38 O O Tabellaria O 0 O 0 0 0 Diatoma 0 0 O O 0 31.03 Cocconeis O 8.33 6.67 4.26 O 24.14 Cocinodiscus O 0 1.67 0 O O Surrirella 0 8.33 1.67 2.13 O 6.90 83 02.4 o oo.~ on.“ o om.o o~.H maomfieoauooo o o wm.~ mo.m o o oh.H mflumflfimnma mm.q an.o mH.m «m.H wo.~ o H~.~ mHHmuapusm mo.HH so.“ mm.a mm.H o o N¢.q ascomaa o om.~ oo.H Na.m oH.oH wo.m mm.q mflfimneso mn.q «N.mu em.m NH.HH as.“ oo.~ 50.x muemasm ca.m mH.m NH.HH om.a ¢N.N Hm.¢ Ho.m mumcooooo wo.m~ mH.m om.HH q~.¢H mH.mN mm.- BH.mN muumfiflmwmum oo.wm um.oo mo.h¢ «m.oq om.o¢ om.~¢ m~.~q «Hagu>mz zoom sand sue soon eumu nuofi .w=< .w:< .w:< mafia kHSH kHSh cowwMMMMJMU muocoo meowumum.HH< wcoa< coHuHmoaaoo zaxmoz Hauoa mo R .mvsum mnu mo mxoma xwm umufim mnu wcwusv cowuwmomEoo maxmo3 cams paw mEOumHv umwuv mo cowuwmomaoo umEE:m::.m mqmwu0< cam: Nam mm~.N mam Hme owe was Nam am New mas -~.H mm Nee mum awn ow mmm mmm mad mes ems Hon mam mH om ma mms man on was mmH o umsws< cam mmo.H me mas “on man man on fins omm mas mom can can oa~.~ mm mN4.N osa.m mma som.a Hoe emm.m Haw.¢ SH mam mae.m mam.w Hmm.ms sea.n Ha.sfiaw A.Sw\8mov mua>wuo< humongous sumuannuu .mes omum .mes ommu .mes oeoa .mum can .mes can m ceauwum m coaumum «a coaumum NH aonumum m acaumum n coaumom .m coaumum .hmoov can vcsouwxomn you vmuoouuou .Hmwuoums 003 no swam you manage pom mucsoo vmuoouuoo aw muu>wuomowvmu cOumnmwuomuu.oH mum ---.. 2001 4‘ \\ \\ ”—‘ uuc/gm A- ‘ \\\ \u \ \ V A\/ 20, 3 I '2 3 4 Weeks 108 Fig. l3.--Comparative Station 8 uptake and loss for Fontinalis antipyretica. Corrected micromicrocuries per gram of wet plant material. 2009‘ 1961 1982 ---s --- ,-””' \. / / \ \ \. \~ / /\\\\\\\ \u\ / \ / 200 ‘/ A \/p uuc/gm 4\/v/ 20 0 2 3 4 57— Weeks 110 Fig. l4.--Comparative Station 8 uptake and loss for Potamogeton sp. Corrected micromicrocuries per gram of wet plant material. 2000« 1959 -------- 1961 ‘——————’ 1982 ---b --- \ \ \ \ \ \ \\ A\ \ \ //\ \ / \ 200 \ ,/ \ \ / \ ,// \,/'/ uuc/gm 20 2 . . ‘ 0 l 2 3 4 Weeks 112 plants collected during 1962. They are comparisons of the trends of activity during 1959 (Knight), 1961 (Bender) and the present study. Generally, the patterns of uptake agree in all three years with the agreement between the present year and 1959 especially good. Differ- ences between the two years, 1959 and 1962, are only in order of magnitude as are the differences between the 1961 data of Bender and the present study. All species except Nasturium officinale exhibit their largest concentration of activity at all stations on July 11, 1962 then pro- gressively decline to a low value between August 6th and 13th. This seemed to be a period of initiation of another increasing cycle of activity accumulation which reached its peak on August 27th. Only Station 8, is illustrated in Figures 12, 13, and 14. Trends for all five plant species during the study were indicative of the initially lower concentrations available at the progressive down- stream stations. The figures also indicate a slow decline in accumu- lated activity for five or six weeks, then a secondary accumulation of activity. Nasturium officinale, the exception mentioned above, had a one week lag in activity accumulation. Other exceptions were on individual dates, i.e. July 30, for Ranunculus sp. and QEEEE.SP° which were eval- uated and found to have a level of tracer accumulation much higher than either the preceding or following weeks. No measurement or machine error was indicated or found by rechecking. The two samples were pro- cessed as usual which includes a washing to remove silt and periphyton, therefore it was presumed that entrapped detritus or invertebrates did not account for the difference, nor did adsorbed materials since the plants were washed with dilute acid before mineralization. I can 113 offer no explanation for the erratic values exhibited. There was a strong indication that fertilization with low con- centrations or low analysis fertilizer can cause an increase of tagged phosphorus utilization in macrophytes for as much as a mile or more downstream within a three month period. The increase was only an in-' dication from the data (Figure 12, 13, and 14) as was the possible increases in biomass with similar fertilization (Figure 15). With .large amounts of fertilizer added or a high analysis fertilizer it would appear that a disruptive pulse of stable phOSphorus was produced which affected P32 uptake in macrophytes adversely. The pulse of stable phosphorus may have been stored in a dense periphyton mat immediately below the point of introduction or it may 'have been stored as a precipitated floc of tricalcium phosphate imme- diately below the point of introduction (Keup, M.S., 1958). The excess P31 may have been cast into new pools or reservoirs which formed and then periodically released some of the stable and some of the tagged phosphorus by unknown methods, with no particular pattern other than a possible correlation with the first major summer rain after the iso- tope introduction. The high concentration of phosphorus (P31) provided by the fertilizer may have made either nitrogen or some minor element a limiting factor which was renewed by runoff from rains. - Figures 16 through 25 exhibit the pattern of tracer uptake by macrophytes during eight weeks of the summer of 1962 and Table 11 gives the specific uptake and Table 12 gives the mean uptake and standard deviations. They all indicate a high initial incorporation of radio- phosphorus followed by a gradual loss for a period of five to six weeks and then a minor secondary accumulatibn of P32. Fontinalis antipyretica 114 Fig. 15.-~Levels of standing crop of macrophytes as related to fertilization. 24,000. 20,000 18,000, lbs/acre 12,000, 8,000. 4,000. Fertilization l—n—-fi Year Pounds formulation Dates 1957 410 21-53-0 Aug. 8-17 July 20-26 1958 230 21-53-0 July 3 1959 300 12-12-12 June 29-July 6 1960 240 21-53-0 June 23-29 1961 none - - _gao none -- - 1959 1980 1981 1962 116 Fig. l6.--Stations 3, 5, and 8 Fontinalis antipyretica activity levels. Corrected micromicrocuries per gram of wet plant material. Allie/fan 500 400 300 200 100 Station 3 25 2 16 723 30 8 13 20 27 Station 5 25 2' 16 13 20 27 50C 400 300 200 Station 8 mu! 2% June 16 July 23 30 8 ~ 13 20 27 Auguuflw 118 Fig. 17.--Stations 12 and 14 Fontinalis antipyretica activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/gm 500 Station 12 4007 300. 200. 100- ‘\ 2'5 '2 9 18 23 30 8 13 20 27 500 Station 14 400 300‘ 200, 100. /\ . , '1--." ; . .. . “h"- 25 2 9 18 23 30 8 13 20 27 June July August 120 Fig. 18.--Stations 3, 5, and 8 Potamogeton sp. activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/gm 500 400 300 200 100 225 Station 3 8 OK 500 400 300‘ 200. 100 Station 5 25 500, 400‘ 300. 200 1001 Station 8 _ \ 25 June 4 1'3 20 27 August 122 Fig. 19.--Stations 12 and 14 Potamogeton sp. activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/gmj 500 Station 12 400 300 200 100 2'5 2 9 18 23 30m 8 13 20 27 500. Station 14 400- 300, 200. 100- 25 2 9 16 23 30 6 13 20 27 June Jul . August 124 Fig. 20.--Stations 3, 5, and 8 Chara sp. activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/gm7 500 Station 3 400 300 200 10c /\ / 25 2 9 16 23 30 6 13 2O 27 500 Station 5 400‘ 300. 2001 100 \,\ \\\/\/ 25 2 9 16 23 30 6' 13 20 27 500; Station 8 400, 300‘ _ 200‘ 25 2 9‘ 16 273 30 8' 1'3 20 27 June July August 126 Fig. 21.--Stations 12 and 14 Chara sp. activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/gm 500J Station 12 400« 300 . 200 . 100 . 25 2' ’ f8 23 30 8 13 20 27 500 Station 14 4001 3001 2001 100% W 25 2 18 23 30 8 13 20 27 June July ' August 128 Fig. 22.--Stations 3, 5, and 8 Ranunculus sp. activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/sm. 500, 400 300 200 100 Station-3 9 16 23 30 8 13 20 27 500 400- Station 5 w #25 9 18 23 30 8 13 20 27 50C 400 (a) O C) 200 100 Station 8 7I: J June 2 9 18 23 30 8 13 20 27 July August 130 Fig. 23.--Stations 12 and 14 Ranunculus sp. activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/gm 500 400 300 200 100 Station 12 25 500 400 300 200 100 a Station 14 June 16 July 30 b' 13 20 August 132 Fig. 24.--Stations 3, 5, and 8 Nasturium officinale activity levels. Corrected micromicrocuries per gram of wet plant material. uuc/gm 500. Station 3 400‘ 300‘ 200. 100' W 25 2‘ 18 23 30 8 13 20 27 5001 Station 5 400, 300; 200, 100, 25 2 9 18 23 30 8 13 20 27 50G Station 8 400L 300 200 100 25 2 9 16 23 3O 6 13 2O 27 June July August 134 Fig. 25.--Stations 12 and 14 Nasturium officinale activity Corrected micromicrocuries per gram of wet plant material. levels. '1'. uuc/gm ' 5“) Station 12 400 300 200* 100, 25 25 2 500‘ Station 14 400, 300 200, ' 100‘ ’,,-- ..-—— 4\______——”"’ 25‘ 25 2 9 18 23 30 8 13 20 27 June July August 136 0.00 m.00H 0.HH «.0 H.Hmm 0.0H 0.H 0.~m 0H H.0N 0.0m m.NH 0.0m m.mm n.0m 0.00 0.Hm w «.00 0.NN H.H0 m.m N.0H 0.00 0.50H w.~m m 0 .am mSHSUcdamm 0.00 m.~m 0.5m H.0 n.0H w.~ 5.0 0.0a 0H 0.00 a.mH 0.0a a.mH H.NH 0.0a 0.m5 5.HOH m 0.00 0.mm 0.00 0.0a 0.00 0.0m 0.0H N.mmN m 0 .mm couoonmuom «.00 N.0N H.NH 5.5a a.mm 0.HH 0.5 m.HN 0H 5.5m «.mm m.5m N.HH m.HH5 5.H0 0.Nw N.N5 w 0.H0 N.0m 0.0H 0.m 0.0 0.00 0.00H H.0m m 0 onGHonmo ESHpSummz --- m.mm 8.8m ,8.Ns a.mmz m.ms --- 5.8m 01 0.00 0.00 0.5m m.w~ 5.Hm 0.Nw m.NOH 5.N0 w 5.00H 0.00H 0.NOH 5.5mH 0.05H 0.00 0.H00 0.0H0 m * mowumummwucm memCMusom 0.00 m.5m 5.mN 0.0H w.mmm N.0H m.~0 0.50 0H 5.00 ~.00 m.m0 N.mH 0.5m N.0m 0.0m m.wH0 w 0.N5 H.Nm 5.05 5.0m 0.0N 0.05 0.05 5.5mm m 0 .am mumso 5N omswsa 8N omsmsa 81 888884 8 08=m=< on 5488 mm 5488 81 5488 as mass unmam u03 mo Emma you mmwusoouowEouofiz .Hmwnouma ucmHa umB mo Emuw pom moproouoHEonowe vouoouuoo 00>Hm commnaum can 00 soamum one: ozu Eoum Hmfinoume uamfim mo mHo>oH muw>wuooowvmmuu.aa mgmoc45H N .884 8884.8 .884 8888.8 .884 8884.8 8884 .48 888888 .804 0050.0 .804 0040.0 .804 0504.0 N004 .5 54:0 mmmmw 000000800om 0440:40com .03 040 00 8040008 50 400 40008 000.0 040.4 04.54 00.00 00.0044 0.0000 04 000.0 000.0 54.0 05.0 40.000 4.0000 N4 500.0 000.0 54.0 00.0 05.400 0.5004 0 000.0 000.0 00.0 44.0 04.0 0.004 0 000.0 000.0 00.4 05.0 00.00 0.000 0 .8000 040008 800040 500\0400\804 500 400 .804 .804 00 00 0040 040: .804 :4 c4 04 0040000 000\NE\8E040 m00\NE\8E040 400 .03 540 0000 .03 040 .03 040 .03 003 .8040008 0000 04 000400000 4000 8=4o008000 000 0040 04:: 400 8043040 4040050040 000000400211.04 04049 141 The area production and rate of production between Stations 3 and 5 was extremely low due to thereduced light, while the area between Station 12 and 14 was one of high light intensity, favorable substrates and currents for the macrophytes. Comparison between the area production rates of this stream and the Red Cedar River near Lansing, Michigan, a warmswater, enriched stream with the same volume of summer flow an average production of 2.6 gm-cal/mz/day shows only the lower most section studied by us, was of the same order of magnitude (personal communication with Robin Vannote, research fellow at Michigan State University). Invertebrates Invertebrates, especially insects, were the major organisms which transferred energy and nutrients from the primary producers to the con- sumers in the West Branch of the Sturgeon River. Generally, the classic trophic system of primary producer, consumer, and decomposer or omnivore was present in the stream. Knight (1961) and Zettelmaier (1961) divided the invertebrates into particular niches for the stream. Using their classification nine invertebrates were studied during the summer of 1962. They included a filter feeder, Simulium sp.; a netspinning caddis, Hydropsyche sp.; two periphyton scrapers or grazers, Ephemerella needhami and g, cornuta; two secondary consumers, Atherix variegata and the fishfly larvae, Chauloides sp.; two omnivores, the pouch snail ghzggpsp.;.and the riffle stonefly, Pteronarcys sp.; and the detritus feeder, Hexagenia sp. Each has been found to occupy a similar niche in studies conducted elsewhere. A single exception to the above classification, Chauloides sp., was found in 1961 when it was found to be an extremely efficient 142 accumulator of radioactive phosphorus during the single year when the P32 was introduced as an organically bound complex (bacteria). The question was not resolved by the present study because with an intro- duction of inorganic tracer again, the fishfly larvae again had only a low level of incorporated P32. The difference in magnitude was striking in view of the head construction and structure neither of which suits the organism to adapt it to feeding on bacteria. Observed feeding habits also contradict the accumulated activity levels present during the 1961 study. Numerous separate observations of the feeding habits had shown the organism to be a secondary carnivore on such organisms as Hydropsyche which they stalked at dusk among riffle substrates. From specimens collected at random, stomach analyses data reveal only insect head capsules were present. It was recognized that normal cur- sory investigation of the intestinal tract does not reveal even large clumps of slime that might contain bacteria. No insect sampled by in- vestigators during the past four years (1959-1962) possessed enough activity in 1961 to contribute the amount present in the 1961 fishfly larvae, yet the same organisms were present in the intestines of pre- served specimens from each year before and during 1961 and in those collected during the present year when the activity level was again low. Either the fishfly was eating some detrital bacteria with its normal food or it greatly concentrated small amounts of the organically bound phosphorus from its food organisms which it had never done before nor did it during the present study. It was a changed form of intro- duction in 1961, but the high degree of loss to the organisms and sub- strate of P32 in the first 240 yards of stream in 1962 would seem to 143 nullify the advantage of changed form. During the present study much more activity was available in the upstream areas and much of its in- corporation was into bacterial forms that were endemic to the stream while in 1961 the tagged bacterial form was foreign to the stream. The activity remained in the upstream areas for a long time with only gradual losses therefore, only a changed form of introduction (bacteria) cannot be the whole answer to this question. In general, the principal mode of tracer accumulation in inver- tebrates is the same as that of fish namely, through the ingestion of food organisms. Radioactive materials are incorporated physiologically into the invertebrate tissue directly. In studies conducted by Robeck, g£_al,_(l954) in the Columbia River it was indicated that radioactivity levels in most aquatic invertebrates were dependent upon the particular organism's metabolic rate and the radioactive material ingested. In this study it was assumed that no radioactivity was contributed by physiological transfer across membranes since no adsorbed activity was washed from the external parts of the invertebrates by 0.01 N hydrochloric acid. Figures 26 through 33 exhibit the pattern of tracer uptake in the order of trophic levels. Hydropsyche sp. was not included because weekly samples could not be systematically collected because of emer- gences that left too few for a weighable sample immediately after the emergence. Also, the replacement organisms had a very different meta- bolic rate and subsequent tracer accumulation. The caddisfly, Brachycentra sp. not previously mentioned had a similar emergence and replacement, but some of the stations were sampled for them after the first three weeks. Their activity levels were initially high, nearly 000 0004 050 000 0004 000 00 05 04 500 0500 0000 000 0000 040 000 000 0 0004 4400 5004 040 0500 0044 050 000 0 0 .08 8004040000 nun- 4004 0000 0500 5045 0500 0400 400 04 0000 0000 0000 0000 40004 00004 0000 0000 0 0000 4000 5000 0050 04054 00004 00000 0000 0 0 0800400 0440405000w 0404 0000 0004 4000 0000 0000 4040 005 04 0000 0050 0000 0000 50044 00004 5000 0005 0 0000 0000 5500 0004 04404 05000 00000 0040 0 0 4800000: 04404080000 0044 000 0044 4004 0000 0000 0500 0000 04 000 0004 0004 0004 0500 0000 0000 0005 0 000 050 540 050 0000 0440 0000 0000 0 0 .08 50440840 88 888884 88 088880 84 umswa< 8 888880 88 84:8 88 84:8 84 84:8 44 84:8 4044080: 80080H 803 00 E040 400 8044000404804042 .0003 00 0040008 00 084>4800 mo 80040> 5084x02 .840>40804 040003 80 40440806 800804 803 00 E040 400 8044000404504048 04 840>04 084>480004004 800804-u.04 m4m x44008< 146 Fig. 26.--Stations 3, 8, and 14 activity levels of the blackfly, Simulium sp. Corrected micromicrocuries per gram of wet insect material. Corrected for background and decay. uuc/gm, 30000 25000 20000 15000 10000 SOOQ Station 3 15 23 30 13 20 27 30000 2500C 2000c 1500 1000a SOOC Station 5 ll 16 23 30 I3 20 27 3000Q 25000 20000 15000 10000 5003 *TV Station 8 16 July 23 3O 6 I3 20 Angust 27 148 Fig. 27.--Stations 3, 8, and 14 activity levels of the mayfly, Ephemerella needhami. Corrected micromicrocuries per gram of wet insect material. uuc/gm 30000‘ Station 3 25000 20000 15000 10000 5000 30000. Station 8 25000 20000 15000 10000 5000 30000 Station 14 25000 20000 15000 10000 5000” m r 11 16 23 30 6 13 2O 27 July August 150 Fig. 28.--Stations 3, 8, and 14 activity levels of the mayfly, Ephemerella cornuta. Corrected micromicrocuries per gram of wet insect material. uuc/gm 30000‘ 25000 20000. 15000 10000 5000 i Station 3 30000 25000 20000 15000 10000 5000 300001 25000 20000. 15000. 100001 5000. Station 5 M T V V' 11 15 23 30 0 13 20 27 Station 8 ‘_ /1/\ il 16 ”231 30 6 13 20 27 July August 152 Fig. 29.--Stations 3, 8, and 14 activity levels of the snipefly, Atherix variegata. Corrected micromicrocuries per gram of wet insect material. 23 30 75090 13 20 '2 / 2500 1500 (H O O Station 8 3500' 2500. Station l4 154 Fig. 30.--Stations 3, 8, and 14 activity levels of P32 in the fishfly larvae, Chauloides sp. Corrected micromicrocuries per gram of wet insect material. July 11 3500. 3000- 25061 20004 1500. 1000 1: -" u U ‘J‘ Station 8 3500 3000. 2500 2000“ 1500 1000 500 July 11 Station 14 16 23 30 Auo o 13 20 27 156 Fig. 31.--Stations 3, 8, and 14 activity levels of the pouch snail, Physa sp. Corrected micromicrocuries per gram of wet insect material. 3003 Station 3 2500.. uuc/gm 1500.. SOC / July 1 Io 25 JO Augxo 13 20 27 3500 Station 3 30 Aug.o 13 20 27 Station 14 '20 27 158 Fig. 32.--Stations 3, 8, and 14 activity levels of the riffle stonefly, Pteronarcys sp. Corrected micromicrocuries per gram of wet insect material. Station 3 2500 uuc/gm‘ 1500 500 ,’ ______ / ’/ ~-~ / -_“,w ‘~‘/ July 11 10 23 30 fiugoc 13 2O 27 “500. J Station 3 25001 1500, 500. July 11 lo 23 30 Aug.6 13 20 27 50. 3 0 Station 14 2500 160 Fig. 33.--Stations 3, 8, and 14 activity levels of the mayfly, Hexagenia sp. Corrected micromicrocuries per gram of wet insect material. uua/gm Station 3 2500 t1500 i 500 r: — -— -— r- — dfnei—r-r'z‘ . . July 1'1 1'5 2'3 30 1,109.0 13 20 27 3500 , Station 8 2500 1500 500 J --.... “3M ; July 11 lo 23 30 Aug.5 13 20 27 350 . 2500 Station 14 162 that of the blackflies, followed by a rapid tapering off of the activity levels. Samples were taken before the isotope introduction and indicated no significant activity above background levels. The two grazing mayflies, Ephemerella needhami and E. cornuta, from Station 3 exhibited the highest levels of accumulated tracer one week after the isotope introduction. Both then indicated a gradual loss of activity during the following six weeks. The blackflies at Station 8 had the second highest levels of P32. The difference between the highest peak of activity at Station 3 for the grazers and Station 8 for the filter feeders was presumed to be due to the difference of feeding habits and the activity incorporation by drift forms at the two stations. Each of the secondary consumers, Atherix and Chauloides, exhibit a lag of accumulation to a high peak level of P32 on the third or fourth week after the introduction. A secondary accumulation follows after a grad— ual loss of P32. The two omnivores, Physa and Pteronarcys, indicate their feeding habits, reaching a high peak of activity on the first week after introduction when it may be presumed that they have been feeding on the periphyton and grazing mayflies while later they reach an equal secondary peak activity when it may be presumed they were feeding on excretory products and more invertebrates than periphyton or its grazing fauna. The lowest levels of activity were as expected in the detritus feeders, Hexagenia sp. A progressively lower level of activity downstream was exhibited as was expected due to the lower level of available activity. No secondary peak of activity accumulation was exhibited in any of the diatom feeders, it was only exhibited by the trophic levels above the primary consumers. This was anomalous since the periphyton was grown 163 on substrates, collected, counted, and found to exhibit a secondary peak of tracer accumulation. This would seem to offer strong evidence of the secondary accumulation coming from organic sources that were precipitated upon the substrates or were ionically bound colloids and not utilized by the primary consumers. Above the primary consumer level the secondary activity increase may only be due to the recycling generated by detritus and omnivorus feeders which were eaten to a greater degree when the other food organisms were less available due to emergence and drift losses. Migration For years it has been known that aquatic stream organisms were normal members of the drift complex moving with the current and along the bottom by saltation due to eddy currents. Several studies have been made relating this drift to the feeding habits of fish (Needham, 1929; Ide, 1942; and Muller, 1954). Dendy (1944) appears to have been the first to formulate the concept that the drifting of stream organisms was a normal process in all streams, even in the absence of floods or adult stream insects and found females with mature eggs flying in a predominantly upstream direction. Muller (1954) proposed the term "colonization cycle" to describe this series of phenomena. In the case of insects and some crustaceans without a flying phase he recog- nized that the question of upstream repopulation was unanswered, he considered this question particularly significant with the crustacean, Gammarus, which has been observed in the drift in very large quantities. With this in mind it was decided to use our tracer phosphorus as an aid to answering Muller's question. The advantages were obvious 164 and the only drawback was that a large series of samples must be counted to determine the background radiation before the tracer was introduced. The methods of isotope introduction in 1962 was such as to effectively create a definite curtain barrier of activity free water from that bearing radioactivity. The curtain was such that anything upstream of the point of introduction that contained radioactive phos- phorus must have flown, swum, or crawled to the point where it was collected. The logical explanation seemed to be a colonization flight, therefore a light trap series using ultra-violet light was setup for collecting adults. The first was 100 yards upstream from the point of tracer addition, and was masked with aluminum foil so it effectively was uni-directional in the upstream direction. Anything attracted to it would have to have flown upstream past it and then circled back or flown downstream. Any P32 carrying specimens would have to have flown past and turned back into it. The second light trap was located at a roadside overhanging cedar about 1/2 mile upstream and was not direc- tional, the third and fourth were located, at irregular times, at successive roadside bridges upstream. Since we had no way of knowing the possible length of flights, nor their intervals, the lights were operated on about every third night. Of thirty sample nights for at least two traps operated for a minimum of eight hours only ten weighable samples were collected after sorting to order. The points of activity levels on given nights are given with distance upstream in Table 16. Samples of adults collected in a Malaise trap which was direc- tional were of little use, presumably because the net was too large 165 TABLE l6.—-Radioactivity levels of adult insects collected by ultra- violet light traps. Corrected counts per minute per gram of insects. Date Type Distance Activity 1961 7-27 mixed 100 yds 88.8 cpm/gm 7-27 mixed 200 yds 85.8 cpm/gm 7-27 mixed 300 yds 19.5 cpm/gm 7-28 mixed 100 yds 3.1 cpm/gm 7-28 mixed 200 yds 42.3 cpm/gm 8-9 mixed 100 yds 0.0 cpm/gm 8-9 mixed 200 yds 39.2 cpm/gm 8-9 mixed 400 yds 14.2 cpm/gm 8-15 mixed 1800 yds 8.7 cpm/gm 8-15 mayflies 1800 yds 105.3 cpm/gm 1962 7-31 mayflies 1800 yds 0.0 cpm/gm 7-31 mixed 100 yds 99.8 cpm/gm 8-3 mixed 1800 yds 23.8 cpm/gm 8-3 mixed 100 yds 138.0 cpm/gm 8-6 mixed 100 yds 151.2 cpm/gm 8-6 mixed 1800 yds 930.6 cpm/gm 8-10 mixed 100 yds 45.8 cpm/gm 8-12 mixed 100 yds 92.1 cpm/gm 8-16 mixed 100 yds 51.8 cpm/gm 8-17 mixed 100 yds 58.5 cpm/gm 8-20 mixed 100 yds 68.4 cpm/gm 166 mesh to collect small mayflies and caddis. ‘The activity accumulated by composite samples of semi-aquatic and aquatic specimens indicated no directed flight pattern nor did weights of the insects collected. Examination of Specimens with reSpect to egg carrying females on one side of the net or the other were also inconclusive of a directed flight. The question of a Species like Gammarus, led us to set up sampling sites at 100 yard intervals upstream from the isotope intro- duction point. They were only sampled as a precautionary measure in 1961, since we had no crustaceans in any of the upper stream study sections and none could be found in the area above the point of tracer addition. A few aquatic beetles and Hydracarina, both of which did not fly, but might move upstream, being large, active, and predatory. Two genera of snails present in the stream were also considered pos- sible migrants since they might also be strong enough to overcome the current, eSpecially along the edges and on logs. In 1961 the isotope was introduced in the form of organically incorporated phosphorus within Escherichia coli 0111. The results of which were reported by Bacon (M.S., 1962) and Bender (1962) who both concluded that the majority of activity was not retained within our study area, but remained tied up within the bacteria and was washed out of our study area which was over two miles long. It was to our advantage that this happened because any activity found in adults or immature specimens that were above the point of isotope introduction, had to have accumulated their activity from the low level of radio- activity that was not incorporated by the bacteria, but was available for incorporation in the stream (about 10 per cent was so available, 167 as reported by Bender, 1962), and was then moved upstream. Among the immature forms sampling was conducted on the same Species upstream of the tracer addition point as those collected downstream of the point. It was found that the immature forms accumulated radioactivity and then progressively moved it upstream for more than 500 yards above the addition point over a period of seven or eight weeks. The data was tested for statistical significance and found to be a significantly greater than background radiation at the 90 per cent level. Further- more it could be shown that the progressive increase in activity was along trophic lines to a culmination in the stronger, large bodied organisms that could be presumed to move more rapidly upstream. The most radioactive species was a stonefly of the family Isoperlidae, an active, large, predatory group. Second in radioactivity levels was the fishfly larvae, Chauloides, also large-bodied, active and con- sidered a predator. Tables 17 and 18 indicate the radioactivity levels present in the respective studies, 1961 and 1962. The difference in timing of the movement was presumed to be due to the difference in mode of isotope introduction. Logically there were two methods of moving the radioactivity up into the area above the point of isotope introduction other than insect flights or crawling immature forms; mechanically by the investi- gators carrying debris on their equipment and possible fish movements. During the 1962 study we again conducted the study, but with the changed method of isotope introduction and form of the isotope. The change from a point source in 1961 to a boom with several point 168 .mmomv cam wcsouwxomn HOW Umuuwhhoo mUCDOU H o o o o o o o o w o o 0 o o o o q 0N mN ma Ndm mm :m nn nu nn nn nn nn nn nn nu nu nn nn n o 0 un nu nn 0 0 .mm manmwmxom mqu o o o o o o o o mmH o me o o o 0 an o Nm 0 Nmmwwuomm 05m nn nn nn nn nu nn nn nn un nn nn n nn 0 nu nn nn nu mo qmmcfiaquOmH No N o o o o o o m 0 5H m o NH m N m o o o N.am mmoumcououm mNN o o o o o o o 0 mm o o o o o o o o «m o ,N.aw mnokwmopvhm mN o o o o o o o o o o o o o o o o moH o o N.mm mwucoomsomum mm o o o o o o o o m mmofi mm o 50 an N N o mmN o {hudm movonSmsu nn nn un nn nu nu nn nn nn nn nn nn n nn nn nn nn nn nu H0 .mw ESHHSEwm 03 q o o o o o o o o o NH 0 m0 mm o 00 o o o Nmumwmwuw> xwum£u< wq N0 om wN HN 0H wq N3 om wN HN 0H 5 ad N0 on mN HN 5H m ammoamm umum¢ mxmn mmmmaom umum< mmmn mmmoamm umum< mmma m>on< mummy oom m>on< mvumw CON o>on< mvumw OOH .oumu mm vmwuooou mum muadoo umsuo H~< .Hm>oH ucwo Hod om osu um unmowmwcwwm was wcsouwxomn cam onEwm coo3uon uasoo aw mucoummmwv mnu Lows: cw mmosu mum wmuaomoum monam> um: vmuomHHoo mmumunmuuo>cH Emmuum mo Aswuw pod ouscwe you .HomH .HH mash commoaou odouOmH .mmmmamu Mmm mo muwm onu Eoum Emmuum H muasoo vmuoouuoov uumcmv muu>wuofluom wo unsoEm unmowmwawwm m on: maowumum mmmsu mo Luon Eouw mmHaEmm .mmumv Ham co vowhamcm oumB mcumh oom can 00¢ um mGOwumum Eouw mmHQEmmm .vasom uo: was zuw>wuom mo uczoew uamowmwcwwm m nan .mmumv Ham co woumfiwcm mums mpumm oom new 000 um mcowumum Eoum mowomam mflsu mo moHdEmmN mnu umuwm want on >ufi>fluom mo unsoem unmowwwcwwm m cm: mwumk 005 um cowumum Eoum monEmm 170 o 03 0 on m m3 m3 533 HNH mm mm 3m NH 3 mom mm 0 ON m3 mu .. .am mmNSm mo 00 NH own NH NH NH «NH mm wH m 3H «H o HmH mHH ON oN 3H NH w chmwmxmm om o wN mN nn NH oH «NH o 05 0H nn NH «H o 5N0 HN mN oH 0N wN muscuoo mHHmquMmmm mm co NH mNN HH O OH NqH o Nm NN mH N HN nn o5 o oHH Nq mH o Hewscmoc mHHmuoEmsmm nn nn nn 0 6H nu nn HHH nn N nn nn HH nn nn nu un un nn nn 0 mmcHumHaomH mHN om mm a mH mm 0N NqH mH 0N NH 5H mN mm qu mo mm NN Hm H3 N .dm mmoumcououm mmH 03 mm mN No mN 3H qu wNH mo nn 0 H3 05 qu mm nn nn Ho mm 5 .am msohmmouvmm Hmu Nm HHH omu 3 03 .. 33 mm 0 mm SH 0 mm mou mm mo Hm NN NN m3 .aw muucmomsomum No 00 NH oN NH nn HN HNH 5w HN mm o o mH m5 wq 0H 0 ON 3 N .mm mmpHoHsmno Nm Hq MN om NN Hq HH ow No 05 mm 5m wH om nu NNH m5 m5 Ho «N 33 .am EsHHaeHm 0N mm MN mH 5m N5 NN wNN 0 ON mm om Nm m5 NH mm H5 N3 mm mH m mummeum> xHumnud wq N5 mm wN HN «H N wq N5 mm wN HN «H N wq N0 mm wN HN «H N mmmonm umum< mzmn mmonmm noum< mmma ommmHom umum< mmmn moHoQO m>on< mpumw oom o>on< mvumw CON m>on< mpumw ooH. .moumu Na pmucmmmuamu mum muasoo umauo HH< .Ho>oH ucmo pom mm.mnu um unmoHMchHm mmz vcsouwxoma cam deEmm consumn mucsoo :H mocmumHMHp map soHSB 0H omozu mum cmucwmmud mmaHm> .NomH .oH NHSW vommmHou maouomH .mmmmHmu Nmm mo muHm man Eoum Emouuma: wwuooHHoo moumunmuum>cH Emmuum mo Asmuw you oudaHE nod mucsoo vmuoouuoov huHmcmw huH>Huo¢nn.wH MHm HH< NO 0 mm me -n 03 33 33 Nm Nm m mm N m3 .am mmNnm HON NO ON OH HN OH ON O3NH NO NN NN 3H OH OH chmwmxmm HO O3 OO OH O NNH OH O NHH ON O 3 O OH muscuoo mHHouoEonmu ONO 3N NNH OH OH NH OO OO O nu O O N NN Hamnvooc mHHmumEmsmm OO O nn OO OH O N NO OO NN O3 un nn nu mmcHHumOOmH OOH 3O O3 N 3O N3 O HO 3O ON OH ON OH OH .Om mhwumcououm OHOH 3O HOH nn O3 NN OH NON O3 OO OOH 33 nu O .Om 050mmmouwmm NONN NOH OO NH HO HO 3O OOH HON OO O nu O3 N .Om muuamommomum OOO Om O3 NH ON ON OH HOO OO OH 3N OH NO O .Om mmOHoHamzo om 3m m3 ON mm mm om mNN on mm mm 33 m3 O3 .am EDHHDEHO HN mm Om ON ON HO O3 ON HO O HH 3 OH OO mumonum> xHuosu< O3 N3 OOn ON HN 3H N O3 N3 OO ON HN. 3H N mmmoHom umum< mOmn mmmoHom uwum< mOma m>on< mcumw OOO m>og< mvumw O03 172 sources was initiated to effectively change the activity pulse from a cone with 3° slopes to a curtain barrier with little or no slope. It was done to create a transect beyond which any insect carrying activity would have to have moved a definite minimum known distance rather than travelled a possible distance governed by the slope of the intermixing cone of activity. A table or graphic representation of the radio- activity levels is erratic because with the large amounts of drift and normal sedentary forms present the probability of always getting radio- active specimens was quite low. As previously stated the Malaise trap was ineffective for aquatic adults and only the data from light traps were used to draw conclusions about the adult movements. A precaution exercised in 1962 was that before entering the area upstream of the point of introduction, the personnel entering the area allowed the current in riffles to wash the debris off their boots and equipment. Mechanical hand washing was effected if needed, when they moved directly into the upstream area from downstream. Other- wise, they moved up to the area only by land and entered with no con- tact with the downstream waters at all. The problem of fish passage of activity was met by shocking specimens in the areas of insect collection and counting them for radioactivity after processing. No activity above background levels was found in any of 125 Specimens of fish of the three major salmonids five weeks after isotope introduction. The longer interval before sampling was allowed for the fish to build up activity after feeding on smaller fish and insects. The fish downstream which have been measured for radioactivity in the last four years have always shown a 173 slow accumulation of activity which was delayed one to two weeks after isotope introduction, presumably due to feeding habits and trophic level changes or transfers of the radioactivity. Robeck, e£_al. (1954) have shown this to be due to the fact that fish take up activity apparently only through the gut, and ingestion was the only means they have of getting it to the gut. If they went upstream after this the feces may have been released and been radioactive. Other workers have shown little or no movement from home areas to be a common salmonid behaviourial pattern, barring Spawning runs in late fall or the early spring. Miller (1954) concluded for cutthroat trout little or no movement took place into or out of several 600 foot sections of stream. Logan (1963) working with cutthroat and rainbow trout had fewer than 40 per cent of his tagged fish move more than 400 feet. In fact, the average per cent of recaptures in excess of 400 feet for all fish within the study section was 6.8 per cent. It therefore was assumed that fish passage of activity could be ruled out. Light traps in 1962 definitely proved that the adults do move upstream in huge flights at different times throughout the summer, dependent upon emergence periods and weather conditions. Again on 30 separate occasions the traps were used, but only captured twelve weighable samples. These weighable samples were always predominantly composed of egg laiden females and were collected as pulses that would fill a pint jar in short periods of time before dusk. Other samples of unweighable numbers collected by hand nets at bridges upstream showed high activity as much as five miles upstream close to the Hoffman Lake source of the stream. Thus quite possibly Muller's colonization cycle was an accurate hypothesis. 174 Immature forms collected in 1962 indicated more activity in more forms and more rapid movement into the upstream areas. Chauloides activity level was practically nil for the first few weeks and some of the omnivores had activity as rapidly as did the herbivores and pred- ators. A possible explanation was that on the night of isotOpe intro- duction there was a large emergence of adult stoneflies and they may have carried some activity upstream in nuptial flights or incorporated in their egg masses. The lack of a time lag at the upper stations above the point of tracer addition may have been the result of this flight or the result of more radioactivity available in all forms immediately below the point of introduction. Thus the organisms didn't have to move as far upstream in 1962, from lower areas where the cone of radioactivity had reached them in the year before. Particular reference to the cone of activity was made because we cannot envision the immature forms moving upstream in areas of full exposure to the currents, instead they appear to move along the sides, along, over, in, and around the numerous logs which studded the area and the weed beds inbetween. A study by Neave (1930) on the mayfly, Blasturus cupidus Say in an intermittent stream revealed similar progressive movements of the immature forms upstream to the extent of as much as one mile, but his conclusion was that they were moving up the intermittent stream in response to crowding and competition for food. He recorded movements as great as 600 feet per day by swimming activity of the nymphs in a current amounting to more than 85 to 110 inches per minute. Or they moved by crawling along the sides and bottom, and when the current was uncomfortable for them they even partially or wholly quit the stream 175 margin, crawling and wriggling forward along the wet border. Fish Samples Systematic sampling of the four major fish species: Salmo trutta fario, Salmo gairdnerii, Salvelinus fontinalis, and Cottus sp., was not a primary consideration during this study, therefore only four collections for tracer levels were made of the fish, beginning on July 16th five days after the isotope introduction. It was found that there was much variation within individual species on a given date as well as between Species of a single collection. Figure 34 is a repre- sentative plot of tracer concentrations of the Cottus. Both Species of sculpin, Cottus bairdii and Cottus cognathus were present and will be referred to here as "muddlers". The individual sample variation was typical of all the fish samples, without regard to species. It appears to be a function of the immediate past feeding history, age, condition, and habitat. Knight (1961) regards the variability as a function primarily of the opportunistic nature of feeding within the four species and the drift composition of the mass passing each fish, Zettelmaier (1961) and Bender (1962) concur in this assumption as does this author. From stomach analyses of the fish used for radioactivity determinations this author would have to agree on both the opportunistic nature of feeding and the drift feeding, but would not place as much emphasis on drift feeding as a general food source at all times. The majority of trout examined had fed upon many bottom organisms that do not drift readily. These organisms would have to be assumed to have been selected and actively picked from the bottom or logs. Two out of every three 176 Fig. 34.--Curve of individual variability within a particular species at a given station demonstrated with muddlers at Stations 8 and 12. Corrected counts per minute per gram of wet fish. 177 unawjfi NN ON NH cowumum m ao3uuum OOH OON com 003 COO OOO OON OOO EO\an 178 specimens that had food in their stomachs contained a species of stone- case caddis which when displaced mechanically, only drifted a few inches before reattaching to the substrate. A few specimens contained several snails of the genus, Ehysag which the investigators could not find readily when sampling specifically for the snails for radioactivity processing. According to Koster (1937) in a New York stream the diet of sculpin consisted largely of larval insects in all stages of development. Bailey (1952) and Dineen (1951) agree with this finding for different types of streams. Knight's work (1961) reveals this to be true of the West Branch of the Sturgeon River specimens. Pentelow (1932) found brown trout to-be largely piscivorous which was not shown by either Knight's work (1961) or the present study which revealed the contents of the stomachs to be mostly insect material with the remainder arachnids. Neil (1938) agrees with the insectivorus diet of brown trout. The discrepancy may be with the size of the specimens sampled in the three streams. Because of the high accumulations of activity in Salmo trutta fario it would seem more probable that the tracer concentrations resulted from an insectivorus diet. Generally, the brook and rainbow trout can be classed as insec- tivorus fish. A regression line for the tracer accumulation by species at the different stations was computed and reveals the difference in concen- tration of phosphorus 32 between trout Species at a given station on a given date. Figures 35, 36, and 37 are the plotted regressions of tracer activity with time at Stations 3, 8, and 14 where comparisons of available water-borne tracer fractions have been made. All the fish 179 Fig. 35.--Regressions of activity with time in days. Salvelinus fontinalis was a negative regression at Station 3. Corrected micro- microcuries per gram of wet weight. uuc/gm 1 HaiflLOW A \O C) C 10 7006 Brown 3000 1000 LT] O C) 100 Station 3 uly'lo 30 August13 27 181 Fig. 36.--Regression of activity with time in days. Corrected micromicrocuries per gram of wet weight. Section 8 plots are given. uuc/gm Rainbow 13003 c 11000 9000 Brown 0 7003 5000 Muduler 3000 1'3300' Statio.: $1 Brook _ 100 . - l//// 16 July 30 August 13 27 Sept. 10 183 Fig. 37.--Regression of activity of fish by time in days. Values from Station 14 samples. Corrected micromicrocuries per gram of wet weight. uuc/gm 15000 Station 3 13000 Brown 11000 a Rainbow 900g Muddler 7000, 5000‘ 30004 1000 , “ ’ 3 . , . .uJuly 30 finayustIJ 20 , Ejept.10 185 were washed with hundredth normal hydrochloric acid and the wash counted for radioactivity. The results of the counted washed showed that no activity was present as adsorbed phOSphorus on the fish at any time during the study. These data reveal no between station difference in rate of accumu— lation as tested by a two-way analysis of variance. They also revealed a trend of upstream concentration of tracer in the rainbow trout, which was followed closely in activity levels by the brown trout, then muddlers, and the least was found in brook trout. This trend was con- tinuous downstream as concentrations built up with time, the only change being more activity in brown trout than in rainbow trout at the lower stations.’ A regression based upon parr would probably be very much dif- ferent, but none was calculated because of the lack of enough samples at each station. When specimens were collected occasionally they had high concentrations of P32 in all species. The much higher concentra- tions in parr and fry has been established by Olsen and Foster (1952) who state that younger, more rapidly growing individuals accumulated relatively greater amounts of radioactivity than the older, more slowly growing fish. They attributed this to a reflection of the more rapid metabolism of the younger fish. In this study two to three times as much was accumulated. Stress has been placed on food chain concentrations of tracer an ingestion by fish because in several types of studies conducted by Davis and Foster (1958), Phillips (1959), and Podoliak (1961) all re- vealed little incorporation of dissolved inorganic or organic phos- phorus by sorption through either the gills or the skin. If one 186 considers loss of tracer through the feces and soluble wastes, a large amount must be so concentrated and returned to the stream in variously combined forms. Podoliak (1961) found indications that in water of 51° F. food which was dyed remained in a trout's stomach up to four days; accumulation in the tissues was slow, but efficient. At higher temperatures the time of contact was less as was the efficiency of removal of phosphorus from the food. The indication was that in 51° F. water and up to 66° F., two milligrams of phosphorus exceeded the digestive and absorptive capacity of the fish and was released only ylittle changed. Fish Biomass and Concentration of P32 Fish populations were determined by using the Petersen formula, from collections made with a 220 volt D.C. shocker. The results in- dicate that brown trout are the numerically dominant salmonid and brook trout are the least abundant (Table 19). By use of the Bailey formula for variance.(Bailey, 1951) the population variance and standard devia- tion were calculated. From the regression slopes the rate of accumulation was inferred for any given time, as well as the total concentration of tracer at that time. Total stable phosphorus was determined by applying the conversion factor obtained by Zettelmaier (1961) for phosphorus per species per unit dry weight. A mean rate of accumulation was calculated, applied to the estimated population per unit area and the total accumu- lation derived. A minimum population was estimated since the samples for estimation were taken late in the summer after some losses to juvenile mortality, cannibalism, fishing pressure, and sampling. 187 TABLE 19.-- Trout population estimates of West Branch and their concen- tration of isotope. 7/16 7/30 8/13 7/16 Station 3 Rainbow Brown Area population 412 563 Area weight in gms. 6226.6 8514.9 if uuc/gm 37.995 114.570 121.295 17.515 Area species accu- 5 5 5 5 mulation by date 2.36x10 7.13x10 7.50x10 1.5x10 Station 8 Area population 1623 1082 Area weight in gms. 19138.6 12759.1 5? uuc/gm 58.367 98.383 488.600 4.679 Area species accu- 6 6 6 4 mulation by date 1.1x10 1.9x10 9.4x10 5.9x10 Station 14 Area population 1228 12213 Area weight 11247.5 111903.5 i uuc/gm 9.562 16.455 308.026 7.004 Area species accu- 5 5 6 5 mulation by date 1.1x10 1.9x10 3.4x10 7.8x10 Total accumulation . 5 - 6 6 . 6 by date in uuc .3.9x10 1.34x10 1.22x10 1.2x10 TABLE l9.--(Cont.) 188 7/30 8/13 7/16 7/30 8/13 Brook 68 1022.7 74.456 54.399 7.224 29.794 5.845 6.34x105 4.6x105 7.4x1o3 3.0x104 5x104 358 4227.4 388.741 461.453 5.849 48.329 270.027 4.9x106 5.9x1o6 2.5x1o4 2.6x105 1.1x1o6 437 4003.4 25.890 304.300 3.064 6.066 11.412 2.9x106 3.4x107 1.2x104 2.4x1o4 4.5x1o4 7.05x106 1.64x10 9.0x105 3.1x106 3.8x107 189 From the calculated values in the Table 19 it is apparent that brook trout are not abundant, and because of their low tracer activity level they do not account for much of the phosphorus transfer within this ecoxystem. Brown trout by gross numerical abundance, weight, and a very high tracer level dominate the production terminus in the stream. Rainbow trout are a close second, but fall off because of reduced numbers of large fish, low stable phosphorus pools, and the period of feeding (diurnal) when insect activity or movement is slight. Rainbow trout seem to have a definite point of growth, beyond which they migrate downstream to larger lentic water areas. For this stream it is approximately nine inches and above. The larger fish are only present during spring Spawning runs and shortly thereafter. Accumulation of activity by muddlers is high and probably a large part of it never goes directly beyond their trophic level. It is assumed that they transfer their incorporated tracer by being eaten by larger trout. They are known food fish and are not large in size so that they don't outgrow their predators. By number alone they account for a tremendous quasi-terminal level of phOSphorus accumula- tion. Closer examination of fish biomass data (Table 19) and tracer levels reveals a series of trends that indicate the fish at the sta- tion located one-third of the longitudinal distance down the stream study section reached a peak of concentration in five days. Downstream levels were a function of the low tracer concentrations upstream. Station 14 accumulations were less, as were those of the upstream stations, 3 and 5, which had a large part of the available activity removed by various means. At the time of the second sampling, 19 days, 190 tracer was still accumulating at all stations within the fish. These tracer concentrations were building up the fastest at Station 8. How- ever, by the third sampling period at 33 days, the tracer was being 'lost from Station 3, the uppermost station samples. It is assumed that adsorbed and incorporated tracer were being recycled and passed down- stream, with continued accumulation evident at the lowermost station. No attempt was made to isolate areas, within fish, that might accumulate more tracer than other areas might. Knight (1961) has a complete description of the level of tracer concentration within a series of fish of each Species. He found that by ranked order of magnitude, bones are highest in activity, followed respectively by the head and gills, viscera, and muscle tissue with the least activity. From a public health standpoint, the amount concentrated in edible fish flesh, i.e. the muscle, was well within the amount given by Donaldson and Foster's (1957) suggested figure of 7 x 10"4 microcuries per gram, being slightly more than half, i.e. 3.7 x 10.4 microcuries per gram of phosphorus 32. I believe that similar values of concentration were maintained during this study period. Light-Dark Study An area of stream was selected for light-dark study of periphyton production on artificial substrates. The first segment was a 200 foot section between Stations 3 and 5 which was characterized by a nearly complete canopy of cedar and tamarack, no higher aquatic vegetation, a riffle over gravel and few logs or large stones for substrates. The mean noon level of sunlight was between 500 and 700 foot-candles de- pending upon the amount of sunflecks striking the light-meter sensing 191 unit directly and the amount of reflected light that was scattered. The second segment was between 400 and 500 yards further downstream between stations 5 and 8. It also was 200 feet long, but it was com- pletely open to direct sunlight and no shore vegetation was present that was higher than one foot high for a radius of 125 feet in all directions. The flow was only half riffle in the sense that the bottom tilted from the left bank to the right facing upstream, and the right side was a deeper run. Numerous logs were present in this section. An open field off to one side of the light area served as a control area and a comparison for the light readings obtained under the water's surface. The clearing was an oval area of approximately 200 feet in a north-south direction and about 280 feet in an east-west direction. The areas were of slightly different flow characteristics, the "shade" zone being quite swift and the "light" zone being swift only on one side. The difference was not presumed significant since the supports for the light measuring units and periphyton substrates in both areas were placed randomly with the aid of a table of random numbers. An equal number of quadrats represented swift and slow habitats in each larger area during the course of the study. The periphyton complex grew upon artificial plexiglass sub- strates attached to a cross-bow on a steel stake. Eight substrates of 1.2 dm2 each, were used to accumulate periphyton for one-week inter- vals, at which time they were removed and replaced by 8 more substrates and the unit moved to a new randomly selected site. Each stake with its 8 substrates had a light meter attached to it during the week interval and all the incident light penetrating to a point one inch above the 192 substrates was recorded continuously by the solenoid recorder. The daily accumulated total readings were recorded each morning between nine and ten o'clock. Maximum and minimum temperatures were taken at the downstream. point of each area. At no time did they differ by more than 3° F. and no consistent pattern of difference was observed. The differences were presumed to be due to available light during some periods heating slow water areas, and to cold ground water contributions to the flow. The substrates were collected in each area and transported to the laboratory in quart jars after the invertebrates were removed. They were frozen overnight and proceSsed for radioactivity the following day. Processing for radioactivity was only slightly different from that of the processing previously described. The 8 substrates were scraped into a common large beaker and the periphyton was suspended in enough 95 per cent alcohol to make the solution up to 50 millimeters. The periphyton and alcohol were allowed to stand for 1 hour, while other substrates were scraped, in a dark container. Then the solution was drawn through a 0.45 micron millipore filter in a millipore apparatus. The filter was wet and pre-weighed. Ten seconds after the filter appeared water free, 3 milliliters of 0.01 N hydrochloric acid were added, followed immediately by a 5 milliliter distilled water wash- The suction was withdrawn after the filter appeared water free again for 15 seconds. The filter was removed and weighed, after which it was placed in a numbered deep-wall planchet. The filtrate was transferred to a Klett cuvette and read using a 660 millimicron filter. Then the filtrate was evaporated to a volume of 2-3 milliliters, added to a deep-wall planchet containing the filter and both were digested with 193 concentrated nitric acid and heat on a hot plate. After complete mineralization the planchet and contents were heated to a cherry red in a muffle furnace set at 6000 C. After cooling, the planchet was counted in the Omni-guard system for radioactivity. Light Measurement A simple, portable, and inexpensive field instrument was built for us by Mr. Clinton Harris, an electronic engineer of Ann Arbor, Michigan. These instruments were built for long continuous operation in the field with a probable error of i_20 per cent. The main requirement was for a system which would measure the total amount of energy falling upon the sensing device over a period of time rather than measuring instanteous intensities. Therefore, the light striking a selenium photovoltaic cell was fed through a transistor, which Served as a matching device and permitted the photovoltaic cell to work through a low resistance. The result of which was a linear response to a wide range of light values in the spectral range which covers the wavelengths involved in biological activity. The selenium cell also allows comparison with the work of other investigators who have used this type of cell. The photovoltaic cell and transistors were packaged as a unit. Temperature considerations were important as related to the transistor Since high ambient temperatures could damage it, therefore, the sensing unit and transistor were packed in a water-tight unit and placed directly in the stream which remains at a near constant temperature. Current from the photovoltaic cell fed through the transistor and resistance was fed into a timing capacitor. The pulsed output from 194 the timing capacitor was directly proportional to the intensity and was sent through a second transistor to energize the solenoid in a five place mechanical counter. The second transistor was silicon to guard against high ambient temperatures since the timing capacitor, transistor, and counter were maintained in a box on shore, as were the batteries which powered the unit. Four 6 volt Sport lantern batteries (NEDA 918) were used to power the 24 volt DC General Controls 5 digit, non-reset counter. Calibration The determination of the lower limit of usefulness of the light and dark units was found to be "dark current" which with proper selec- tion of the component parts could be made such that the device was re- liable down to a light level of well under 1 foot-candles incident directly on the photovoltaic cell if a reasonable counting rate was employed (2 counts per second). By inherent limitations the device stopped counting with a little under 0.1 foot-candles directly incident upon the photovoltaic cell. Because selenium cells are non-linear at high light intensity and because light intensities in excess of 1000 foot-candles damage the cells it was decided that a maximum of 500 foot-candles should be incident upon the photovoltaic cells at any given moment. A range of l to 200 foot-candles was considered ideal and by filtering the cell Should be representative of the stream. To reduce the intensity of the incident light upon the cells neutral density filters were employed. Interchangeable filters were 'provided for the "light" and ”shade" units. 195 Filters with a factor of 50 (density 1.7) were used in the "light" units. Thus the incident light range of 50 to 10,000 foot- candles resulted in a unit incident light on the photovoltaic cell of from 1 to 200 foot-candles. In "Shade" areas where lower light levels were important in terms of their aggregate contribution, advantage was taken of the fact that very high light intensities were not expected. Therefore, filters with a factor of 5 (density 0.7) were used in the Shade units. In order to reduce the error resulting from angle or incidence the photovoltaic cell and the filter were mounted and sealed against moisture behind a glass hemisphere with a finely ground surface. The counting interval was established as about 1 second for a light intensity of 200 foot-candles incident upon the cell. The devices were calibrated with the sensing unit immersed in water of the same temperature as that of the stream in which they were to be used in order to eliminate errors arising from the temperature characteristics of the photovoltaic cells and their matching transistors. .Figure 38 is a photo of a complete unit without the battery and counter housing (a wooden box). Table 20 is a representative series of instantaneous light readings taken at 20 foot intervals downstream from the upstream edge of the dark area. The levels of incident light were measured with a Weston light cell which measured the light on an upper and lower scale with a range on the scale from zero to seventy foot-candles. The readings tables were on the lowest scale setting and were read directly. They are noon readings on a Slightly hazy day when maximum open field light was 7000 foot-candles from the same meter. 196 Fig. 38.--A light sensing unit in its immersible covering, with glass hemiSphere and cord to Shore counter and power source. 11 x... l : o>og -.80\0q._ z m 2 55322 .oflmnoq E 3353an 23m 5032: mooEom nozmgomfl meegog OESGOHOSN 198 TABLE 20.--Noon light readings in foot-candles, as measured by Weston 756 light cell. Taken at 20 foot intervals (longitudinally) on a slightly hazy day. Distance From North Stream Bank, in Feet Transect (Right Bank) 3 6 9 12 15 18 21 1 20 30 20 5 5 10 - 2 36 42 42 42 4O 35 30 3 25 25 30 30 30 26 - 4 10 35 45 45 36 15 10 5 35 3O 28 28 15 10 5 6 10 30 25 20 12 10 - 7 25 ' 30 5— 1 3 5 - 8 40 40 25 10 6 4 - 9 70- 70- 70- 70- 60 56 55 10 60 70 7O 60 55 50 45 Light reflecting off water--5 foot-candles. Open sunlight 7000 foot-candles. 10 A.M. readings averaged 5-10 foot candles less. 3 P.M. readings averaged 10-15 foot candles higher. 199 Area Comparisons The total light intensity values of replicated data from randomly located stakes from the two Sites (light and dark) were computed by a matched observation method, the results of which are given in the appen- dices. These tests indicate that the two areas were significantly dif- ferent at the .99 per cent level in the total light received. From the same stakes the substrate weights of periphyton and radioactivity accumulated during given intervals were measured and the weight of periphyton versus radioactivity tested for the light and dark area by a covariance analysis modeled after that given by Walker and Lev (1953). The results are given in the appendices and indicate that the two Slopes are not significantly different, i.e. the rates of accumulation in both areas were the same. A test of covariance of the intercepts given in the appendices indicated that the intercepts were significantly dif- ferent at the .99 level, i.e. the rate of accumulation in both areas was the same, but the amount of activity present from measured time intervals were different between areas. The light to dark area rela- tionship was one where the higher weights present in the dark areas had less radioactivity associated with them at any given time than did the light areas. It would appear then that the ratio of bound phosphorus to wet weight of periphyton of similar Species in the two areas differ, with the dark area generally having a lower value than the light area. With the rate of weight accumulation and radioactivity accumulation the same in the two areas (light and dark) the lower amount of phOSphorus com- pounds present at the time of measurement must be due to some type of organic phosphate degeneration. This may be due to the significantly 200 different amounts of light energy reaching the two areas. The light area receives nearly full light (95 per cent) while the dark area re- ceives on 13 per cent of full light. The lesser intensity in the dark area may be compensated for by more of the light being of long wave- lengths due to the canopy filtration favoring passage of the red end of the spectrum, but it apparently does not compensate by increased chlorophyll production, with associated phosphorus uptake. Increased efficiency of production may account for the difference in production on the substates since it is known that the rate of photosynthesis will increase with the light intensity at lower light levels up to a certain level of light intensity after which it rapidly falls off. Maximum efficiency is often reached at only one-third to one-fourth the peak intensity (Meyer and Anderson, 1952). These factors must enter into the area production just as the fact that phosphorus deficient land plants tend to store excessive sugars and inorganic nitrates rather than proteins with their asso- ciated phospholipids and nucleoproteins. If it can be assumed the same physiological and biochemical mechanisms and processes are present in aquatic plants as in land plants then the above factors may account for the differences in organic weight accumulated in the two areas, but not the difference in radioactivity. The production of more material per se would have the same incorporation rate normally, as it does, and a higher total incorporated amount of phosphorus should re- sult. The difference must lie in the total energy received in the two areas, they are known to be significantly different with the dark area receiving much less external light energy. Therefore, the energy re- quired must come from stored products through the energy cycle utilizing 201 ATP and ADP and all the intermediate stored phosphorus compounds. The energy demand for the high rate of production must be met by a high respiratory rate. Calvin and Benson (1950) have shown that phospho- glyceric acid is both an anabolism and catabolism constituent and may well be a respiratory waste. The intermediate phosphates which were stored are tranSported to new building Sites and used to build more phospholipids and nucleic proteins with organically bound phosphate constituents. During the building the stored phosphates are changed from the old forms to new forms for reincorporation by the high energy system utilizing ATP and ADP and the total result is a recycling of stored phosphates for the energy they can supply when the external en- vironment does not supply enough energy for maintaining the photo- synthetic rate. With respect to the possible difference in efficiency of pro- duction between the two areas the work of Emerson and Arnold (1932) may well explain the observed differences in the West Branch of the Sturgeon River data. They exposed cultures of Chlorella to intermittent illumination at the rate of 50 flashes per second, the periods of illumination being much shorter (0.0034 second) than the intervening dark periods (0.0166 second), and obtained a photosynthetic yield per unit of light which was increased by about 400 per cent as compared with the rate in continuous light. Assuming the photochemical reaction comes first, the results were concluded as follows: when illumination is continuous the products of a light reaction are formed faster than they can be utilized in a relatively slower dark reaction. When the light is intermittent, all or most of the products are removed from the photochemical reaction, and the photosynthetic output per unit of 202 light is considerably greater. They measured the dark reaction and found it to proceed in less than 0.04 second at 25° C., and to be greatly influenced by temperature. The light reaction takes place at a Speed of about 0.00001 second and is unaffected by temperature. With low light intensities and adequate carbon dioxide, a photochemical reaction is limiting and temperature will have little effect on the rate of the process. With high light inten- sities and adequate carbon dioxide, but low temperatures, the rate of photosynthesis is limited by the dark reaction, and will increase con- siderably with a rise in temperature. Considering the stream as a whole the water is well aerated and rich in carbon dioxide as well as being uniformly cold. The shaded areas were well interspersed with light flecks which changed instan- taneously and continuously. This would well fit the conditions described by the above authors and in the light areas the dark reaction may well be limiting, whereas in the dark areas the photochemical reaction may be the limiting factor due to reduced energy, but still greater in terms of the amounts of products produced. SUMMARY The form of tracer available to the organisms of the ecosystem was Shown to be an important factor in the time and amount of phos- phorus take up by living and dead materials. Uptake by the sediments was high within the experimental zone, with larger amounts being found in the biologically incorporated State than in the adsorbed State. These differences increased with distance downstream from the point of application. The particulate fraction and the organic fractions of the water were Shown to be very important to the exchangeable phOSphorus that may be recycled as it moved downstream. The amount of tracer incor- porated in organic fractions increased in direct proportion to the in- crease of the producer organisms. Bacteria were shown to incorporate as much tracer as the diatoms when both were endemic in the stream and in the same volume of water, but the magnitude and rate of diatom incorporation appear to be more important to the transfer of inorganic phosphorus to the trophic structure of the Stream than that of the bacteria. Mbvement of radio-activity was demonstrated in connection with the terrestrial and aquatic phase of the large, mobile aquatic insects, and to movements of snails which moved readily upstream even against rapid currents. Distances of 500 yards for the crawling forms and five miles for the flying forms were recorded. Fish movement was very 203 204 limited and accounted for very little dispersal of phosphorus. The volume and number of invertebrates drifting downstream was high and contributed considerably to the downstream movement of the isotope. Light controlled the productive structure of the stream where it was on the border line of the duration and intensity required for peri- phyton growth. The levels of light were so low in the areas having a complete vegetative canopy that the food reserves built up during certain intervals of peak illumination were called on to supply the energy for continued maintenance at the low efficiency periods which occurred during much of the daylight hours. LITERATURE CITED Bacon, James P., Jr. 1962. Translocation of radiophosphorus in a stream by bacteria and inorganic ions. Master's Thesis, Michigan State University. Bailey, N. T. J. 1951. On estimating the size of mobile populations from recapture data. Biometrika 38:293-306. Bailey, Jack E. 1952. Life history and ecology of the sculpin, Cottus bairdii punctulatus, in southwestern Montana. Copeia, 4:243-253. Bender, Michael E. 1962. The bacterial translocation of radioactive phosphorus through a lotic ecosystem. Master's Thesis, Michigan State University. Blum, John L. 1957. An ecological study of the algae of the Saline River, Michigan. Hydrobiologia 9(4):36l-408. Boroughs, Howard, Walter A. Chapman, and Theodore R. Rice. 1957.' Laboratory experiments on the uptake, accumulation, and loss of radionuclids by marine organisms. Nat. Acad. Science - Nat. Res. Council Publ. No. 551:80-87. Butcher, R. W. 1932. Studies on the ecology of rivers. II. The miCroflora of rivers with Special reference to the algae on the river bed. Ann. Bot. 46:813-816. Calvin, M,, and A. A. Benson. 1949. The path of carbon in photo- synthesis IV. Science 109:140-142. Chase, Grafton D. 1960. Principles of radioisotope methodolOgy. Burgess Publishing Company, Minneapolis, Minn. 286 pp. Clifford, Hugh F. 1959. Response of periphyton to phOSphorus intro- duced into a.Michigan trout streams Master's Thesis, Michigan State University. Correll, David L. 1958. Alteration of the productivity of a trout stream by the addition of phosphate. Master's Thesis, Michigan State University. ~ Davis, J. J. and R. F. Foster. 1958. Bioaccumulation of radioisotopes through aquatic food chains. Ecol. 39:530-535. Dendy, J. S. 1944. The fate of animals in stream drift when carried into lakes. Ecol. Monographs 14:333-357. Dineen, Clarence F. 1954. A comparative study of the food habits of Cottus bairdii and associated species of salmonidae. Amer. Midl. Nat., 46:640-645. 205 206 Donaldson, Lauren R. and Richard F. Foster. 1957. Effects of radia- tion on aquatic organisms. From the Effects of Atomic Radiation on Oceanography and Fisheries, Publ. No. 551, Nat. Acad. of Sci. Nat. Res. Council, 96-102. Einsele, W. 1941. Die umsetzung von zugfuhrten, anorganische phOSphat in eutrophen see und irhe ruchwirkungen auf seinen gesamthaushalt. Fischerei. 39:407-488. Grzenda, Alfred R. 1956. The biological response of a trout stream to headwater fertilization. Master's Thesis, Michigan State University. Grzenda, Alfred R. 1960. Primary production, energetics, and nutrient utilization in a warm-water stream. Ph.D. Thesis. Michigan State University. Hayes, F. R. and J. E. Phillips. 1958. Lake water and sediment. IV. RadiophOSphorus equilibrium with mud, plants and bacteria under oxidized and reduced conditions. Limnol. and Oceanogr., 3:459- 475. Holden, A. V. 1961. The removal of dissolved phosphate from lake waters by bottom deposits. Verb. Internat. Verein. Limnol. 14:247-251. Horton, P. A. 1961. The bionomics of brown trout in a Dartmoor stream. J. Animal Ecol., 30:311-338. Ide, F. P. 1935. The effect of temperature on the distribution of the mayfly fauna of a stream. Univ. Toronto Studies, Biol. Series, No. 39. . 1942. Availabiligy of aquatic insects as food of the speckled trout, Salvelinus fontinalis. Trans. No. Amer. Wild. Conf. 7:442-450. Keup, Lowell Edward. 1958. Biological Responses of Fertilization in a lake and stream. Master's Thesis, Michigan State University. Kinsman, S. 1957. Radiological health handbook. U.S. Dept. of Health Education, and Welfare, Public Health Service. Robert A. Taft San. Eng. Center, Cincinnati, Ohio. 355 pp. Knight, Allen W. 1961. The translocation of radiophosphorus through an aquatic ecosystem. Master's Thesis, Michigan State University. Koster, Wm. J. 1937. The food of sculpins (Cottidae) in central New York. Trans. Am. Fish. Soc. 66:374-382. 207 Krumholz, Louis A. and Richard F. Fosrer. 1957. Accumulation and retention of raiieactivity from fission products and other radio materials by freshmwater organisms. Nat. Acad. Sci.-- Nat. Res. Council. Publ. No. 551:88-95. Labaw, L. W. , V. M. Mosley, and R. W. G. Wyckoff. 1950. Radio- active studies of the phosphorus metabolism of Escherichia coli. Jour. Bacteriol. 59:251-262. Logan, Sidney M. 1963. Winter observations on bottom organisms and trout in Bridger Creek, Montana. Trans. Amer. Fish. Soc. 92:140-145. ' Lund, J. W. G. 1950. Studies on Asterionella Formosa Hass. II. Nutrient depletion and the Spring maximum. Ecol. 38(1): 1-350 Miller, R. B. 1954. Movements of Cutthroat Trout after different periods of retention upstream and downstream from their homes. J. Fish. Res. Ed. Can., 11(5). Muller, Karl. 1954. Investigations on the organic drift in north Swedish streams. Rept. Inst. Freshwater Res., Drottningholm, 35:133-148. Neave, Ferris. 1930. Migratory habits of the mayfly, Blasturus cupidus Say. Ecology 11(3):568-576. Needham, P. R. 1929. A quantitative study of the fish food supply in selected areas. N. Y. Cons. Dept., Suppl. to 17th Ann. Rept. p. 192-205. Neill, R. M. 1938. The food and feeding of the Brown Trout (Salmo Trutto L.) in relation to the organic environment. Trans. Roy. Soc. Edinb. 59:481-520. Olson P. A. and Foster, R. F. 1952. An incident of high mortality among large rainbow trout after treatment with pyridylmercuric acetate. Pentelow, F. T. K. 1932. The food of the brown trout, (Salmo trutta L.). J. Animal Ecol., 1:101-107. Peters, J. C. 1959. An evaluation of the use of artificial substrates for determining primary production in flowing water. Master's Thesis, Michigan State University. Phillips, Arthur M., Jr. 1959. The known and possible role of min- erals in trout nutrition and physiology. Trans. Amer. Fish. Soc. 88:134. 208 Podoliak, Henry A. 1961. Relation between water temperature and meta- bolisms of dietary phosphorus by fingerling brook trout. Trans. Amer. Fish. Soc., 90:398-403. Robeck, Gordon C., Crosswell Henderson and Ralph (1 Phalange. 1954. Water quality studies on the Columbia River. Robt. A. Taft San. Eng. Center. Cincinnati, Ohio. Rodhe, Wilhelm. 1948. Environmental requirements of fresh-water plankton algae. Symbolae. Bot. Upsalienses, 10, No. 1, 149 pp. Roos, Tage. 1957. Upstream migration in adult stream dwelling insects. Inst. of Freshwater Research, Drottningholm. Rept. No. 38. Waters, T. J. 1961. Standing Crop and Drift of stream Bottom Organism. Ecology 42(3):532-537. Walker and Lev. 1953. Statistical inference. Henry Holt & Co., Inc. New York. Welch, Paul S. 1948. Limnological Methods. Blakiston Co. Whittaker, R. H. 1961. Estimation of net primary production of forest and shrub communities. Ecology 42:1 Jan. 1961. Young, 0. W. 1945.. A limnological investigation of periphyton in Douglas Lake, Michigan. Trans. Am. Micr. Soc. 64:1-20. Zettelmaier, John L. 1961. The translocation of radiophOSphorus through a lotic ecosystem. Master's Thesis, Michigan State University. APPENDIX Expansion of data presented in thesis. Y- radioactivity (P32) X1 - light X2 - weight Y x1 x 2 384.9 399040 .0255 142.0 114515 .0925 603.2 985500 .0285 0.0 109918 .0240 0.0 1075932 .0175 451.4 112735 .0125 309.0 490060 .0270 322.2 109987 .0385 247.3 700255 .0520 401.8 108464 .3050 161.8 929244 .0241 322.3. -LlZfleél 3.92% 3215.8'5314815 .7595 300.0 2.9.15.2 3035.6 X1 980592 78006 1564066 247032 2088576 149624 1285472 136030 1446384 105506 1056936 _105363 10233577 x2 .0335 .0290 .0415 .1030 .0375 .0285 .0320 .0696 .0880 .0932 .0475 3.0.81.1 .6843 733.7 176.1 408.8 324.4 71.9 146.6 307.7 300.0 309.1 198.2 170.0 350,8 3497.3 X1 880000 127734 1106180 259590 1089296 122176 458936 95577 955144 104185 633920 ”122.121 5952851 AhEA light .0255 334.9 .0330 400.2 . 570 733.7 .1150 1513.8 dalk .0923 142.0 .0293 327.5 .0520 175.1 .1175 045.0 light .0235 503.2 .0415 42.0 .0303 403.8 .1300 1054.0 dark .0340 0.0 .1030 414.7 .0790 324.4 .2000 739.1 light .0175 0.0 .0375 313.9 .0355 71.9 .0905 335.3 dark .0125 451.4 .0235 152.2 .0405 140.0 .0375 750.2 light .0270 309.9 .0323 300.0 .0325 307.7 .0915 917.0 dark .0335 342.2 .0095 310.4 .0420 300.0 .1500 952.0 light .0020 247.3 .0330 197.5 .0145 309.1 .1045 753. dark .3050 401.3 .0932 75.3 .0020 193.2 .5202 075.3 lignt .0241 131.3 .0475 300.0 .0341 170.0 .1057 031.8 dark .0425 300.1 . 811 201.2 0 8 350.8 &L710 22212 sum x .7596 3216.8 .6843 3035.6 .5637 3497.3 Sum x2 .1527 .0476 .0296 1269*55.6 924642.6 1389806.9 (x)2 .5770 .4683 .3178 11687609.1 9214367.4 12231107.3 141‘ .0481 .0390 .0265 n 973967.4 707905.0 1019258.9 x2 .1046 .0086 .0031 295733.2 156736.9 320548.0 Treatments 1 240.0 grand totals " 2 169.7 x Y " 3 112.7 x 1.9516 9877.5 583.4 x2 .4718 8992650.2 , (x)23.8087 97565006.3 (x fin, .3174 8130417.2 KIN/\Afi r\ Matched observations test of significant difference between obser- vations. l 2 light 339040 930502 9a5500 1534020 1075932 2033573 490030 1235472 750250 1445364 ’92iipt. 1051293) dark 114315 I30,. 109913 247032 112735 143024 109937 130030 108464 105503 112464 105333 X 15662725 3 ; 2543737.R t ; §_;_g ; 3.49 with 11 df. Sd Therefore: significant at .99 level I 330000 1100130 ‘r‘r' .~.’ 3".) ‘ 4w. \IQ J4.tu 955144 3101734 035920 2020100 % 113} _ __~ _~____ difference area lign‘ 127734 320355 1339377 14.17 259553 015540 3029190 10.91 122170 364535 3858969 9.04 95577 341594 1893374 15.25 104151 318121 2843663 10.00 120.41 337908 2282132 12.90 x2 44303458904219 X :13.06% 1412-2521’é..."044 4 25%;»... n 0 SS 419370/0000000 _85 2870788900000 n-l b S 574157700C00 S 757731.9 .rld .. VJ 1.; f)' 40' Covariance analysis of SlOpCS of Leignt vs. raoioactivity in the light vs. oarK areas. 5 x2 xy y2 N 12000.0 .0005 3.3 77705.9 3 355.7 .0075 3.5 194C3.4 3 -3oea.7 .0003 - 2.2 lo2420.4 3 5303.0 .0033 17.5 95121.9 3 9000.0 .0003 2.7 54033.9 3 ~1C200.0 .0005 - 5.1 00313.4 3 0.0 .0000 0.0 54.0 3 500.0 .0005 - 0.3 950.7 3 -l4o4.3 .0023 - 4.1 0551.4 3 379.5 .0550 43.? 54330.0 3 5333.3 .0003 1.0 12022.1 3 ~31ll.l .0009 --2.3 11401.7 3 Test of whether 12 5'5 are significantly different from one another. . .2 . ‘ . ‘r‘. r. rind sum of o apout eacn legICSSlon Ly: o‘ z y‘ --b xy .\) \1 \1 6 \CJ I \1 \I. (A ‘. O C I I H C ‘4 .L\.. 1...; 1o2423.4 -- 3033.7 ; 1:4359.7 ’ 951210\9 - 24d'3205 : 2319.4 54055.9 - 24300.0 3 29733.? Covariance analysis of slopes 01 Leignt vs. Iauioactivity con't. U Q I O (3 5%.0 3.3.7 - 133.: ; 73;.7 JEJbloq - V‘L):J.\J - 2Q708 xx \ o‘)ind _: 223914.0 11431.7 - 3711.1 - 2750.0 b .2 2297.2." A: - o - Chg. “1.1.2.2.. 92:3... 2. 94708 average 2x1 + ......1th ”2 .0720 2 of. - (n-k-l) - (’o- 2-1) ; 23 of. , 2 2 zkzd) £231 + "°°°+E\ll; " 1v éxyl * 000*2Xy ) average ' (JVCIC’U'E: ; 554301.3-947. (33.0) ; 5543Cl.3 --05393.2 ; 432903.6 2 2 unbiaseo estimate of sijma 2 _-_-_ 2 _—_-2£o)in_g: _-_ of ind ’-— " -. J ' 2 c‘ 2 or) 'x" ’ 0"" ' - - uncidseo estimate of Sigmal : 31 -; £94103.c - 173/3.2 ; H7032jr4_ ll 11 F 27 n E .087 11_d£& d 403 l 12 of. bus :1 I 2.72 with 11/12 CCVLG€S of freedom J 576 leve Therefoze the shepes are not significantly different. Analysis of covariance: Test of intercepts that are assumes to be equal, adapted from Walker and Lev (1953). Peripnyton weignt vs. iauioactivity sum X sumXY sch sam X2 sum Y2 i. light .1150 05.04 1513.3 .0050 840023.7 .0337 uaxk .1175 31.79 345.0 .0121 115343l.5 .0392 light . 300 43.40 0&».u .0001 532721.7 .0433 aark .2000 03.3n 733.1 .0174 2772ll.5 .0037 light .0905 1%.32 335.0 .0033 103702.3 .0302 dark .0875 10.80 751.2 .0031 243418.4 .0292 light .0915 27.97 217.0 .0023 230717.3 .0305 dark .1500 47.53 952.0 .0031 31157o.3 .0502 light .1545 37.19 753.9 .0118 l95700.4 .1734 cark .5202 105.90 375.5 .1453 20639b.b .0352 light .1057 23.95 031.3 .0040 145079.2 .0352 mark .1713 45.97 352.0 .0117 253884.3 .3142 bum (sum X) - 1.9515 Sum(sumx¥) - 530.23 Sumksum X2) ; .8299 Oumlsum Y2) ; ’550479.7 "fir .: 1.1x :_ 1.22.1.9. - .0242 _ N 35 YT ; 11.2 : 2L7? :2. ; 274.4 N 30 average Uetermine adjustéeumegns by \/ 1 _'_'_ YT - 121‘ ()KT - X1) (A) (J) ;;_: C2.) 0.) (A) Covariance of 274.4 274.4 274.4 274.4 274.4 274.4 274.4 274.4 274.4 274.4 274.4 274.4 947.8 -94703 947.8 947.8 947.8 947.8 947.3 947.8 947.8 Calculate one Y;YT-bT'(xT-x intC2cepts con't.-- (-0117‘4) grano regiession equation pooling all data. 1) where: :- §§§22§_;_§§5,4Z Z Q0,Zg__: 4 .2299 -. 058 .1241 C_ H I I H) H .'\) l C.“ ._; % y—i l l C O I). 9945.5 - 425.1 (52.75) - 5527517.2 F .: 21.152.12.22 - 44340229. ll __2 1----12 zmgzgissli ; 27.08 with 11222. 21273.4 23 of. gab424.2 23 intercepts ale significantly miffeicnt at the .999 level. Three-way analysis of variance f0: gates, groups, insects during 1901 immature movement study. A: da es 7, 14, 21, 28, 30, 42, and 48 days I: - 54130;)?» - fitngi‘ix, sis-4.6;;- L ; distance - 100, 200, 300, 400, anu 500 yaius Source dun of at mean squaies squaie m 4220.40 0 704 40 b 7440.20 5 1488.05 C 4801.10 4 1200.23 AB 23339.31 30 794.54 50 21429.31 23 1071.49 A0 25071.17 24 1039.03 ABC 100309.12 120 835.91 no significance fiom any source at alpha :quotient .05 and uistance in 'T1 0.843 1.780 1.435 0.951 1.282 1.230 Light - 399540 985500 1075932 49oooo 750255 929244 114e1s 109913 112735 109937 108454 112454 DJ with oark matched observations 980592 55 050 2033575 1235472 1440334 1055930 73000 247032 149024 880000 1100180 1039295 453931 955144 flf.A{ Significant at .99 Revel 4253524 2240453 3101734 Anon n AUZUlU' r\.,-. \‘ 347 \ L; J r L) W 415540 ’. 1 1C"; 1 J41 1v )4 3379o8 sum X - SUI“ X2 - (2.2-113 n 2 SS : -19.... .: n-l S2 - s - ) 1939077 3029190 384535 3808930 1591274 2a43aa3 2282132 _ 15302725 _ 44303452904219 251020044425525 ,. U -m--—_ I f O 574157700000 _ 757731.9 Test of heteroyenity of 1692888100 between («I Area UP XL 1 11 .1040 2 11 .0030 3 11 .0031 Total 33 .1103 Total reduced sum of squares accounts for tne reouceo some of squares for each area degrees of freecom. differences between Therefore: of Total 32 Areas 30 Uifference 2 XY 42.2 VA 1—4 |\) O‘- \1’ c1 m () kw O (.— 773073.1 Remaining M q u m (,0 (IN. J5 (,1 H (0 OLA) (DI-~-I -< X 0.} 02) l\) 0) U" (_n (A) O -. \O (11) \C I U‘ (.3 r0- 0 a l 01‘ areas. .0 2070.0 49430.0 UégIGOS of freedom represen t the 3 1891885100 coefficients. H15 24121.4 oegrees of only 3 x 10 _-_ 30 UP 10 10 10 32 Replicates 110 55 x 334.92 :__,__.__.__.._:___L,04;;__>g___30;_,._1.1 - _1_,__9_5;15 :3; 9377.5 F“ N (A), j \JU eror 52.9 - Q-40o.7) - 0.3 3 452.) . ~ ‘ .4 Of. 2; k? 4y 'r b-X byx y ya2 Replicates 12 -.00o5 -400.7 -1900731.g Treatmed.s 12 .0070 0.0 - 50057.7 . 709.2 4515.2 05272.9 Error 33 .1020 452.0 2545474.5 2475.9 1120592.3 1724531.5 Treatment plus error 45 .1905 458.0 2784815.9 3245.1 1125207.5 l790154.7 Test significance of regression error SS of MS E Regression ll20592.3 ll lf1372.0 1.89 Error l724881.8 32 53902.5 Test significance of theoifferences between area means (treatments). Treatment plus l790154.7 45 error Error l724881.3 32 53902.5 0.15 Difference 55272.9 8 8919.1 Adj . Treat- ments 4015.2 11 Be -Bt 50557.7 Simultaneous S Mean values of the four replicates are microcuries per milliliter Left ><| 0.0193 0.0193 0.0183 0.0015 0.1035 0.8555 2.8228 2.0790 1.5290 0.2500 0.0275 0.0088 R ; mean activity 8 ; standard deviation "V 5 0.0084 0.0059 0.0123 0.0184 0.0343 0.2070 0.2527 0.5035 0.1225 0.1807 0.0429 0.0080 across amp feet from the rignt tank. 0.1015 0.0273 0.5245 0.1048 0.1383 0.4578 1.2840 1 .0305 1.5250 0 .4473 0.1033 0.1223 les were taken at 4 feet from the left bank, of water. 0.5558 1.0110 1.6545 1.0418 0.0475 0.1240 0.1903 the center, stream samples from 4 repeated transects. given in terms of micro- Right S 0.058 0.1032 0.0200 0.0593 0.1481 0.1097 0.2914 0.1718 0.1992 0.0305 0.0023 0.1895 ano 4 {31,9 flaw 051‘: 0.1.1 ”'Clliilnjlljiflljflll [1111111111111 fill“