PRODUCTEON AND NUTRIENT REMOVAL ‘ BY MACROPHYTE COMMUNITIES- [N ARTIFKSIAL STREAMS Thesis for the. Degree of M. S. MICHIGAN STAIE UNWERSITY GLENN A. DUDDLES 1967 TH EEEE LIBRARY \\ WWW L ,# W. .sc... WM 3‘2 39°36-_ W ABSTRACT PRODUCTION AND NUTRIENT REMOVAL BY MACROPHYTE COMMUNITIES IN ARTIFICIAL STREAMS by Glenn A. Duddles Production and nutrient removal relationships were stud- ied for three macrophyte communities in two continuous flow artificial streams. The streams were constructed for outdoor study to simu- late natural growing conditions. The artificial streams were established with water from the Red Cedar River which was stored in a large holding tank. Con- trolled enrichment in the form of phosphate and nitrate was added to stream A during two separate study periods. Daily upstream- downstream measurements were made to record nutrient removal within the system. The macrophytes selected were Ceratophyllum demersum (L. ), Vallisneria americana (Michx.) and Elodea canadensis (Michx. ). These were gathered from local stream areas and planted in the streams on an artificial wire mesh substrate. Glenn A. Duddles Community production measured by increase in dry weight was doubled in the enriched environment. There was not a noticeable difference in production between 4 mg/l and 2 mg/l enrichment levels. The Ceratophyllum community had the greatest cropped production (3. 7 g dry wt/mZ/day) and the Elodea community the greatest cropped plus uncropped production (5. 4 g dry wt/mZ/day). The phosphate level was reduced by an average of 18. 7% and the nitrate level by 19. 5% in the artificial streams. A greater percentage of organic phosphorus and nitrogen was found in the macro- phyte communities of the enriched stream. The average protein value of these communities was 19% including a maximum of 25% The N/P quotient for all the communities was relatively constant at 4. 5. PRODUCTION AND NUTRIENT REMOVAL BY MACROPHYTE COMMUNITIES IN ARTIFICIAL STREAMS By Glenn A. Duddles 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 1967 ACKNOWLEDGMENTS I offer my sincere thanks to Mr. Vernie A. Knudson for his supervision of this study, for his guidance. as my advisor, and for his unselfish contribution of time. I wish to thank Drs. Niles R. Kevern and K. L. Schulze for their time given as members of my guidance committee, and Dr. Peter I. Tack for his helpful assistance in the initial stages of my graduate study. I also thank Dr. Gerald W. Prescott for his aid in taxonomic problems. My appreciation is extended to many of my fellow gradu— ate students for their discussions, opinions, and suggestions relating to the study. To my wife, Marilee, I give my thanks for her patience and understanding through the years of study, and I express my sin- cere appreciation to my parents for their continual encouragement. I also am grateful for financial assistance from a re- search assistantship under the Michigan State University Agricultural Experimental Station. ii TABLE OF CONTENTS INTRODUCTION MATERIALS AND ME THODS Artificial Streams Flow Design . Substrate Aquatic Macrophytes Nutrient Addition . Community Production Daily Tests . PhOSphate . Nitrogen Light, Temperature, and pH RESULTS AND DISCUSSION . Study Conditions Community Production Individual Sections . . Total Macrophyte Communities Bottom Deposits Nutrient Observations Phosphate . Nitrogen . Organic Nutrient Levels SUMMARY LITERATURE CITED . APPENDICES iii Page 10 10 13 13 15 15 16 16 17 17 22 23 26 29 29 29 31 35 4O 42 45 Table LIST OF TABLES Identified algae from the artificial streams Macrophyte community production by individual sections for period I and II Total community production for each macrophyte community over the 30 day study Total deposited bottom material for each macro- phyte community over the 30 day study . Average nutrient level and percent removal during period I and II Percent organic phosphorus and nitrogen in cropped material from period I Percent organic phosphorus and nitrogen in cropped material from period 11 . Average organic nutrients and percent protein for each macrophyte community . iv Page 18 24 27 30 31 35 36 38 Figure LIST OF FIGURES Photograph of the entire artificial stream . Photograph of the feed line assembly . Diagram of the sections of plants within streams A and B Photograph of the chemical feed pump assembly . Page 12 14 Appendix I. II. III. IV. LIST OF APPENDICES Daily pH, temperatures, and environmental conditions for period I Daily pH, temperatures, and environmental conditions for period 11 Original starting plant mass and formulas for developing uniform origin Daily nutrient levels and percent removal for period I and 11 vi Page 45 46 47 50 INTRODUCTION Eutrophication is defined by Ruttner (1953) and Hutchinson (1957) as the natural process of lake maturation. This process pro- ceeds at an infinitely slow rate under natural conditions. However, in recent years there has been increased alarm over an accelerated aging of many of our lakes and a degradation of our streams. This change is a direct result of pollution connected with civilization. The first evidence of these problems occurs with the appearance of nuisance growths in the form of algae and higher aquat- ic plants. Such growths inevitably interfere with the municipal and recreational uses of the water. A major source of the problem is excessive enrichment of natural waters with nutrients essential for plant growth, mainly nitrogen and phosphorus. In any nutrient problem, careful consider- ation must be given to the major sources which contribute the nutri- ents. A partial list of these would include nutrients from sewage, sewage effluents, industrial wastes, land drainage, applied fertil- izers, precipitation, urban runoff, and release from bottom sedi- ments. The greatest contributor of nutrients from this list would be that from sewage and sewage effluents. For many years it was not uncommon for untreated sewage to be regularly discharged into rivers and lakes. The ramifications of this practice involve more than merely enrichment problems, and as a result of public interest, this type of pollution has been largely eliminated. Most sewage is now treated or at least partially treated before discharge into receiving waters. However, in terms of enrichment, sewage treatment does not solve the problem involving eutrophication. The mechanical and biological sewage purification procedures used today remove only a small part of the effective nutritive material for plant growth. Or- ganic phosphorus in the sewage plus simple and complex phosphates from synthetic detergents result in phOSphorus concentrations far above the requirements for plant growth. Treatment or organic wastes re- sults in an abundance of nitrogen in all forms. Ammonia and nitrite are transient forms and generally are oxidized to nitrates upon de— composition. Mackenthun (1964) reports secondarily treated sewage effluent from the Madison, Wisconsin, metropolitan area of 85 square miles with a population of 135, 000 to annually contribute 8. 5 pounds of nitrogen and 3. 5 pounds of soluble phosphorus per capita. This extreme enrichment has a tremendous effect on the growth of aquatic vegetation. Mackenthun (1965) indicates that submerged aquatic plants could be expected to produce at least 7 tons per acre containing 32 pounds of nitrogen and 3. 2 pounds of phospho- rus per acre. This represents a significant removal of nutrients from the water by aquatic plants. William Beck (1962) removed 30 to 90% of the total nitrogen in a treated sewage effluent at Orlando, Florida, using beds of algae. Oxidation ponds utilizing plants and algae for treatment have become widely used in small community operations. Bartsch and Allum (1957) studied the variability in design and loading for this method of treatment in the northern plains area. Although this treat- ment works sufficiently for stabilization of raw wastes, it does not eliminate the enrichment problem. The increased mass of algae and plants within the pond die and in time become a source of nutrient release within the ecosystem itself. Loehr and Stephenson (1965) discovered that the nitrogen and phosphorus content of a treated efflu- ent was generally increased after time in an oxidation pond used as tertiary treatment. The purpose of this study was to measure the effect of nitrogen and phOSphorus enrichment on three higher aquatic plants and, conversely, the effect of the plants on the nutrient concentration in terms of nutrient uptake. This was done utilizing two separate 14 day growth periods during August 1967 at East Lansing, Michigan. The three plants selected were Ceratophyllum demersum, Vallisneria americana, and Elodea canadensis. These were chosen on the basis of their availability and variety of growing nature. All three are vascular, submerged aquatic macrOphytes found frequently in enriched environments. The CeratOphyllum, often referred to as coontail, was included because experiments by Schulze (1966) indicated that it had the greatest active production in a similarly enriched sit- uation. MATERIALS AND METHODS Artificial Streams A number of studies have been made concerning produc— tion by higher aquatic plants in natural and laboratory conditions, but none have incorporated the use of an artificial stream. Odum and Hoskin (1957) studied the production of microorganisms in an artifi- cial laboratory system. Stokes (1960) and Kevern (1962) measured primary production by periphyton in a completely recirculating artifi- cial stream. The classical method for measuring growth by higher aquatics such as that used by Rickett (1922), Schuette (1928), or Pen- found (1956) entails random cropping from a given area in a natural situation. A certain degree of error is inherent in growth measure- ments of this nature. Wetzel (1964) points out the importance of the rooting portions of macrophytes in determining production. Natural situations make accurate collection of entire rooted macrophytes ex- tremely difficult. To minimize these problems, an artificial stream was constructed for this outdoor study to enable better control over the substrate area and certain growing conditions. The channel dimensions were 104 feet in length, 2 feet in width and depth. The channels were constructed from 3/4 inch exte- rior grade plywood sheets attached in 8 foot lengths with wood screws. All joints were reinforced on the inside with 1 X 2 pine stripping and additional screws. Further strengthening was attained with a liberal application of Weldwood Plastic Glue to all fitted joints. This water insoluble glue also aided in making the channels watertight. The eight foot sections were placed together upon the flat concrete edges of two large parallel reservoir tanks. These parallel units were then connected by two additional sections supported on a wood framework making a U-shaped channel out of 13 eight foot sec- tions (Figure 1). All of the sections were firmly joined together using 1 X 6 pine boards with wood screws and sealed with glue. Crossbars were placed across the open top of the channels at four foot intervals to keep the sides from spreading. A wall was constructed in the midpoint of the base, sepa- rating the structure into streams A and B. Throughout the study, stream A was used for experimentation while stream B was main- tained as a control. The interior of the streams was coated with two applica- tions of Glidden Nu Pon Coat Epoxy Resin Paint which yielded a dur- able waterproof finish. All exterior surfaces were coated with a high quality exterior oil base paint. Figure 1. Photograph of the entire artificial stream. Flow Design The streams were constructed so that water pumped in at each side of the separating midpoint would flow to the ends of each stream in a continuous non-recycling pattern. The water was dis— charged after overflowing the desired 18 inch depth at the end of streams A and B. The rate of discharge was in this way directly controlled by the incoming flow. The water supply for this study was pumped from the Red Cedar River into the large storage tank alongside stream A. This was done with a large irrigation pump and 5 inch aluminum pipe. The storage tank was replenished as was necessary during the study period. A Little Giant Submersible Pump, model #12NR was used to pump water from the storage tank into both streams. A 3/4 inch rubber hose attached to the pump was divided beyond an overflow shunt into feed lines for each stream. The overflow shunt and both feed lines contained compression valves to control the incoming flow (Figure 2). Throughout the study, the incoming flow was maintained at 1. 2 gpm which established a 16. 5 hour retention time within streams. This flow was regularly checked with a volumetric con- tainer and a timer. Figure 2. Photograph of the feed line assembly. 10 Substrate In order to minimize the sorption, storage and release of phosphorus and nitrogen by substrate particles, an artificial substrate of 1/2 inch galvanized wire mesh was used in the streams. The mesh was formed into eight foot lengths and placed into each stream four inches from the bottom. To insure no contamination, the wire mesh was thoroughly coated before being placed into the channels. This was first attempted using the same epoxy paint which sealed the inner surfaces of the streams. However, this non-flexible coating cracked and chipped at the slightest stress on the wire. The mesh was then covered with "Flexiblac, " a water insoluble paint which dries to a flexible, yet completely inert finish. Aquatic Macrophytes The plants used in this study were Elodea canadensis, Vallisneria americana and Ceratophyllum demersum. All were gath- ered from local streams and stored until planted in the artificial streams. At that time, each plant was hand picked to eliminate any damaged portions and wet weights were obtained for the plants placed in each section of the streams. The plants were separated into the six equal eight foot sections within streams A and B. These sections were numbered 1 11 to 6 beginning at the head of each stream. Both streams thus con— tained two separate sections of each type of plant (Figure 3). All of the Vallisneria was collected from the Grand River below the Waverly Road Bridge south and west of Lansing. These plants grew in abundance there scattered among others and were in excellent condition. Each plant was placed individually through the wire mesh with its root stock downward. This procedure, although tediously slow, had satisfactory results with minimum loss after planting. The Elodea came from the Red Cedar River near the Zimmer Road Bridge west of Williamston, Michigan. It was planted in small clumps with the rooted portions pushed through the mesh 3 or 4 inches. Within a short time the clumps spread laterally, form- ing dense beds. The Ceratophyllum also came from the Red Cedar River with half of it collected below the Zimmer Road Bridge and half at Ferguson Park in Okemos, Michigan. This plant had to be threaded down through the mesh and back out with both ends then extending up- ward. This procedure was necessary because of the non—rooting nature of this plant and was best accomplished with the wire out of the water and the plants placed in spaced rows. 12 I I 1 C I I 1 C 2 2 V V 3 3 E E 4 4 Stream A Stream B C C ‘—T 5 5 V V 6 6 D '1' E E ‘P D I = Incoming Flow C = Ceratophyllum V = Vallisneria D = Discharge E = Elodea Figure 3. Diagram of the sections of plants within streams A and B. 13 Nutrient Addition The line of incoming flow into stream A passed through a small pump house at which point a concentrated nutrient feed solu- tion was added. This was done with an electronic impulse regulated diaphragm feed pump from The Precision Chemical Pump Corpora- tion, model #1201-1. This unit allowed accurate nutrient addition in amounts as small as 5 ml/min (Figure 4). The nutrients added in this manner consisted of phosphate as the dibasic salt, KH P04 and nitrate nitrogen as potassium ni- 2 trate. The phosphate and nitrate concentrations were maintained at 4 ppm during the first growth period from August 1 to the 15th and at 2 ppm for the second period from August 18 to the Blst. Community Production Community production was measured by the increase in dry weight during the two week growth periods. The plants in all sec- tions were cropped at a uniform height on the morning beginning a growth period. This was done utilizing a jig designed to rest on the sides of the channel with extensions down into the water and an ad- justable horizontal edge. This adjustable edge was set at a level 6 inches above the wire mesh substrate and slowly moved down the stream while all plant material in contact with the edge was clipped l4 Figure 4. Photograph of the chemical feed pump assembly. 15 with hand grass cutters. This procedure was repeated at the end of the growth period and all cut material in each section was collected and wet weights obtained. The croppings were then dried in an oven at 105°C for at least 24 hours, after which they were cooled in a des- sicator and constant dry weights taken. At the end of the final growth period, all plant material was stripped from the artificial substrate. This, along with all other organic material from the bottoms and sides of each section, was weighed and dried for analysis. Daily Tests Water samples were taken at the inflow and outflow of each stream every day of the testing period. These samples were analyzed and the differences recorded daily. Phosphate Ortho-phosphate was measured daily using the ammonium molybdate-amino napthol sulfonic acid method as described in. ”Stan- dard Methods for Examination of Water and Waste Water, " APHA ' AWWA ' WPCF ' , 1965. Total organic phOSphate determinations were made on the cropped plant material and the final bottom scrap- ings following the appropriate test outlined in Standard Methods. The 16 dried plant material was ashed in the furnace and uniformly mixed before testing for total phosphate. Nitrogen The Brucine Test was used to measure nitrate nitrogen while ammonia nitrogen was tested by Nesslerization. These proce- dures were followed as outlined in Standard Methods. Nitrite nitrogen was periodically checked using prepared powders deve10ped by the Hach Chemical Company. The Bausch and Lomb Spectronic 20 was used to measure color in each case. Total Kjeldahl nitrogen deter- minations 'were made with a semi-micro method applying an Aminco system and a Sargent Spectro-Electro Titrator. Conversion to raw protein was made using a standard factor of 6. 25. Light, Temperature, and pH Each time samples were taken, the air temperature and the water temperature were recorded. General observations of weather conditions and amount of light reaching the plants were noted daily. A Beckman Zeromatic II pH instrument was used to obtain pH readings. RESULTS AND DISCUSSION Study Conditions Several distinct stages of plant condition were evident during the study period. Soon after the initial planting into the chan- nels, most of the plants supported a dense growth of periphyton. This was a mixture of several species of algae which covered a major por- tion of the plants and channel walls. This first bloom seemed to reach its peak several days after the introduction of the river water supply and remained a factor throughout the study. These algae are generally associated with enriched habitats. (Prescott, 1962). Near the end of the first growth period, a second bloom of Oedogonium _sp. appeared in both streams and became the domi- nant algae. There was a continuous flow of fluffy clumps of this plant to the surface and out of the system. The third algae of any significance was the filamentous Cladophora glomerata which began in the first three sections of stream A half way through the second growth period. Prescott (_o_p. _c_i_t_.) reports this algae is normally associated with cement walls. It probably came into the artificial stream from the large storage tank (Table 1). 17 18 Table 1. Identified algae from the artificial streams.* I Mixed Periphyton H In Scenedesmus alternans Oedogonium _s_p. Cladophora glomerata S. bijuga S. dimorphus S. quadricauda Pediastrum boryanum _13. duplex Palmodictyon viride Hormidiumi gp. Stigeoclonium 3p. Mougeotia _sp. Coelastrum microporum Misc. Diatoms Fragilaria crotonensis Melosira granulata Cocconeis sp. Synedra EE' *Algae identification by G. W. Prescott 19 The Elodea was planted in spaced clumps which soon spread laterally to form dense beds. An initial die back within these beds was followed by evident new growth. Long roots appeared all along the plant and particularly in the area beneath the wire mesh. Because of this attachment to the artificial substrate, very little Elodea floated away due to the failure of the plants to attach to the substrate. During the first growth period, the bloom of mixed algae was particularly evident as periphyton on lower portions of the Elodea. New shoots of growth remained relatively free from the periphyton and the attached algae was less prominent along the edges of the channel where shading from the sides occurred. This was due mostly to the fact that the Elodea grew better in the shaded side areas than in the Open center. Blackburn (1961) reported that high light intensities have an inhibitory effect on the growth of Elodea canadensis and that this ef— fect follows a brief period of initial active growth and results in even- tual death of the plants. It was evident that some Elodea was gradually dying as the study progressed. Since both streams were completely unshaded, this may have been due to high light intensity. In one case, section 6 of stream B, it was necessary to replant fresh material before starting the second growth period. Since Vallisneria freely roots to varied substrates, this plant was expected to thrive in the artificial streams. However, this 20 is the one plant which failed to show significant growth during the en- tire study. At the time of planting, all of the large exterior leaves which were damaged or browned were stripped off so that only fresh live material was placed in the streams. Throughout the entire study, the large leaves deteriorated and continually broke loose and floated to the surface. The Vallisneria. did not attach to the artificial substrate as expected and periodically whole plants floated to the surface. These were returned to the mesh or recorded as lost material from the re- spective sections. While making the final stripping of plants from the streams, it was discovered that the Vallisneria did not die entirely. The meri- stematic portion of each plant had been very active and a network of lateral stolons and new growth was evident beneath and just above the mesh. This growth was not measured in the cropped material for a growth period but was included in the total production at the end of the study. The Ceratophyllum was by far the most successful plant in this experiment in terms of growth and plant condition. It grew rapidly and remained relatively free from periphyton. Wilkinson (1963) report- ed the Optimum conditions for Ceratophyllum growth to be high light in- tensities at 30° C temperatures. This is substantiated by the success of the coontail in the similar conditions of the artificial streams. 21 Due to the non—rooting nature of Cerotophyllum, there was some difficulty in keeping this plant attached to the substrate. The slightest stress on the plants caused the slender stems around the wire to break. When the bloom of Oedogonium _sp. reached its peak near the end of the first growth period, the mass of the algae caused much of the coontail to break loose from the artificial substrate. It was for this reason that sections A-4, B-1, and B-4 were stripped and replanted with fresh material before starting the second growth period. No further loss of Ceratophyllum was experienced and all sections were in good condition at the termination of the study. The overall environmental conditions may have had an effect on the differences between growth period I and II. Period I from August 1 to the 14th was generally clear sunshine with very warm temperatures. On the other hand, period II from August 18 to the 3lst was characterized by numerous overcast days and consider- ably cooler temperatures. Merkle and Fertig (1963) describe an average pH differ- ence of 0. 88 between a rainy, overcast day and a bright sunlight day. This is largely a result of a shift in the HC03-CO3 equilibrium caused by photosynthetic processes. This is also evident in the pH gradient established as the water passes over the plant material from head to discharge within the artificial streams. The difference in stream A was an average of 0.27 for the first period and 0. 31 for the overcast 22 second period. A maximum change of 0.5 occurred in both periods. The pH fluctuation in this study was apparently limited by the buffer- ing capacity of the supply water (Appendices I and II). Community Production The term macrophyte community production was used in this study since no attempt was made to separate the higher aquatic plants from attached periphyton. Production was measured by the increase in material during a specified time interval. Naturally the actual amount of increase was directly related to the amount of orig- inal plant material placed in each section. To validly compare the growth between sections and streams, some method must be used to relate the growth back to the original stock. This was done by using a weight to weight percentage. Since the only measurement possible on original stock plant material was wet weights, all forms of growth are related by weight percentage back to the original wet weight planted. Due to the fact that not all the growth was recorded in the cr0pping procedure and some plant stock was continually being lost, determining a set value for original starting stock presented several problems. This was further complicated by the fact that some sec- tions were restocked with fresh material between the two study peri- ods. 23 A series of formulas was designed to establish this start- ing value, taking into account the increases and decreases throughout the entire study. These formulas were used to determine all per- centages expressed concerning growth and production (Appendix III). Individual Sections The results in Table 2 show that the production in the treatment stream A was consistently greater than in the control stream B. The range of production expressed in terms of percentage of in- crease (wet weight) from original starting mass was 78. 5% maximum and 3. 2% minimum for stream A compared to 39. 2% maximum and 1. 6% minimum for stream B. The only instance where a section in the control had more growth than the correSponding treated section involves section 6 in the second growth period. Here the non-treated Elodea had more production than the treated. This may be attributed to several reasons. Section B—6 was the only Elodea section which was restocked with fresh material before beginning the second period. As Blackburn (op. cit.) pointed out, Elodea canadensis responds under these conditions with an initial Spurt of growth and then slowly dies off due to high light intensity. While this was going on in B-6, the Elodea in A-6 continued its already downward progression. It is noteworthy that the growth during the second period was consistently greater than that of the first even though the 24 5m 2:... is a; v.2: «.mm «mom 2: s; «d: .2: as; :33 N m 9% 92 m; 98 23 3m 92 a; 0.3 2: saw a o; mm: as ms 3. as S 3: 5o :2 9... 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This indicates that it may be possible to reach a level of enrichment beyond which production is no longer increased. It is more likely that in the absence of minimum nitrogen and phOSphorus concentrations, the effect of temperature, light and other environmental conditions becomes more prominent in influencing production. Total Macrophyte Communities In terms of cropped production, Ceratophyllum was the most productive of the three plants in the study (Table 3). This is most evident in the cropped plant material because the coontail had less unharvested growth. In all cases, the treated plants substantial- ly out—produced the non-treated. The greatest difference in this re- spect was in the Elodea when considering total cropped and uncropped production. This indicates that the Elodea exhibited the greatest lat— eral growth in the lower portions. Merkle and Fertig (1963) showed that Elodea canadensis grew more rapidly near the bottom of a 2 foot deep pool exposed to high light intensities. The difference was from 0. 89 cm/day increase in length at the bottom and 0. 18 cm/day at the top. In view of previous discussion, this was probably due to the dif- ferences in light intensities at these levels. Even though the Elodea had more lateral uncropped growth than the others, when the results 27 NN H..H --- ..... N H NN NHVNH NHL. momma. Nd Hie --- ..... Hio m. N NH .N NNH «26:9:me N .N N .N o .H. H. .NNH N .N N .H. S .N N .NHH 8318888 m Emmfim N .N Him N .N N .HHN N .N N N 3.2 H .HNH a H .H H. .o N .H N .HN N .N N .o NN .N N .HN «289:; N N N .H. N .N N .NHN H. .N N .N N .N N .5: 83338.88 < Ewofim Ham». umml 45> HQ El 55 HQ Imlhuoq «2 :HwHHO vommofiocb + N2 HHHwEO NPRCBQ QOHU BO :3 Bo m Ho .N no.5 H33. Blow Ho ON .N N. .353 amp cm on: p95 hficaflfioo mfnnmogomfi 30.8 no.“ :ofiosponm bHHHsfiaoo HwHoH .m 3an 28 are interpreted on the basis of increase from original starting mass, Ceratophyllum was the greatest producer. The differences in the condition of the plants should be noted again. The Elodea grew at first but then tapered off continu— ously throughout the study while the Ceratophyllum remained in good condition. Hasler and Jones (1949) demonstrated that the more active higher aquatic plants had less attached algae. Some unknown mecha- nism inhibited phytoplankton production. It is probable that this is a result of light shading by the growth of the macrophyte. As a result of the plant condition, Elodea was covered with more periphyton than the active coontail. This may have been a significant contributing factor in the greater portion of uncropped growth from these sections. Coontail also had the lowest percentage dry weight from wet weight plant material (7. 3%). This results from higher water adherence to the coontail plants causing the wet weight value to be greater'and the corresponding percentage to be lower. The values obtained for cropped production are directly dependent on the procedures used in cropping. Due to the high vari- ability inherent in such procedures, the significance of the values relate only to this particular study. The values eXpressed as total cropped plus uncropped production are valid for comparison in any situation of similar conditions. 29 When considering total production under enriched environ- ments, the Elodea community produced at a rate of 8. 7 tons dry wt/ acre/year. This was 3.2 times as much overall production when compared to the control stream B. Bottom Deposits At the beginning of the study period, the bottoms of the artificial streams were completely free from any deposited material. At the conclusion of the study, the detritus on the bottom of each sec- tion was collected and weighed (Table 4). This detritus appeared to be mainly decomposing plant material. In all cases there was more detritus deposited in stream A than stream B. The Elodea was the greatest contributor of bottom deposits, followed by Vallisneria and Ceratophyllum respectively. The Elodea community deposited bottom material in the enriched stream A at a rate of 18. 1 tons dry wt/acre/ year compared to 11. 8 tons dry wt/acre/year in stream B. Nutrient Observations Phosphate During the first growth period, the phosphate concentra- tion was maintained at an average concentration of 4. 07 mg/l in stream A. This concentration of soluble phosphate entering the 30 system was reduced by an average of 0. 73 mg/l before flowing out of the channel. The highest reduction during this period was 1. 6 mg/l and the lowest 0. 3 mg/l (Appendix IV). Table 4. Total deposited bottom material for each macrophyte com- munity over the 30 day study. Total g g Dry Wt. g DW D g DW D Ton DW Wet Wt. Bottom —— 2 acre . g WW P M —— Planted Deposns —— year day Research Stream A Ceratophyllum 1746 302 - 0.17 3. 4 5. 4 Vallisneria 2728‘ 677 0.25 7.5 12.0 Elodea 4355 1014 0.23 11.3 18.1 Control Stream B Ceratophyllum 2887 202 0.07 2. 2 3. 5 Vallisneria 3882 502 0. 13 5. 5 8. 8 Elodea 3659 669 0.18 7.4 11.8 DW = Dry Weight DW D = Dry Weight Deposited WW = Wet Weight WW P = Wet Weight Deposited In contrast, the control stream B remained at an incom- ing phosphate concentration of 0. 5 mg/l throughout both testing 31 periods. The reduction of phosphate concentration within the control was on the average 0. 1 mg/l. The incoming concentration of ortho-phosphate was low- ered to an average of 2. 5 mg/l in stream A for the second growth period. This brought about a corresponding lowering of the average phosphate concentration reduction within the system to 0. 42 mg/l. The maximum reduction was 1.0 mg/l and the minimum 0. 3 mg/l during this test period (Table 5). Table 5. Average nutrient level and percent removal during period I and II. PO4 Reduct. % NO3 Reduct. % ppm ppm Removal ppm ppm Removal Growth Period I StreamA 4.1 0.7 17.9 3.2 0.8 25.3 Stream B 0.5 0.1 20.0 0.4 0.1 25.0 Growth Period II StreamA 2.5 0.4 16.8 2.1 0.5 21.0 Stream B 0.5 0.1 20.0 0.4 0.1 25.0 Nitrogen The ammonium concentration remained at a level below 0. 5 mg/l in both streams throughout the entire study. Likewise the 32 nitrite concentration remained at an insignificant low level during the study. These concentrations were constant within the artificial stream situation. These transient forms of nitrogen are readily con- verted to the more stable nitrate in natural waters in the presence of oxygen, which was the case with the Red Cedar River and the large storage tank. The nitrate concentration incoming to the control stream B was constant at near 0. 4 mg/l during both growth periods. There was an average reduction of this concentration of about 0. 1 mg/l for both periods (Table 5). During the first growth period, the nitrate level was held at an average concentration of 3. 16 mg/l for stream A. This level was lowered in the system by an average of O. 8 mg/l for the first 2 week period. The greatest reduction was 1. 3 mg/l while the lowest was 0. 1 mg/l. The second growth period had an average incoming nitrate level of 2. 1 mg/l with an average reduction of 0. 44 mg/l. This in- cluded a maximum reduction of 0. 7 mg/l and a minimum of 0.2 mg/l. The soluble ortho—phosphate concentration coming into the enriched stream A was reduced by. an average of 17. 5% in passing through the channel. Similarly the nitrate concentration was lowered by 23. 1% within the stream. These reductions, although measurable, 33 are not as great as was expected for this study. Certainly the great- est factor limiting the removal of nutrients in this study was the con- dition of the plants. The photosynthetic activity of the plants was not great enough to counteract the buffering effect inherent in the water supply used. As a result, the pH was never raised to the point where active phOSphate precipitation took place. Also a continual decompo- sition of plant material occurred, phosphorus was probably released within the system. The retention time within the stream directly influenced the percentage of nutrients removed. If the retention time had been increased, it is likely that more nutrients would have been removed. At the waste treatment plant in Orlando, Florida, William Beck (pp. gi_t.) showed that there was a direct relationship between retention time and the percent removal of nitrogen and phOSphate in a tertiary treatment pond. He indicates the maximum removal to occur at re- tention times from 2 to 2. 5 days, which is significantly longer than used in this study. His results show nitrogen removal up to 90% under controlled conditions. At retention times of less than one day, his nitrogen percentage removed was in the range of 20 to 30%. Loehr and Stephensen (op. cit.) show that over long periods of time, the nitrogen and phosphorus concentrations may increase within tertiary treatment ponds. This is largely due to the release 34 of nutrients from decomposition of material within the system. If a means of regularly harvesting the production was applied to biological tertiary processes, it is likely that significant removal of nutrients could be accomplished. However, Bogan (1960) points out that there has been con- siderable difficulty in harvesting unicellular algae. Numerous attempts at harvesting algae incorporating screening, settling, centrifuging, and chemical coagulation have failed in developing an efficient, eco- nomical method. In comparison, the nature of higher aquatic mac- rophytes enables more efficient cropping. A number of mechanical harvesters have been developed to aid in the control of aquatic plant growths. Organic Nutrient Levels All of the macrophyte communities in stream A had higher percentages of organic phosphorus and nitrogen than those of stream B. (Tables 6 and 7). This agrees with Wetzel (op. cit.) in citing Gessnar and Kaukal (1952) about the uptake of phosphorus by Elodea being proportional to the concentrations present in the water. The crOpped material from the first section of Cerato- phyllum in stream A consistently had higher percentages of nutrients than the succeeding sections. This difference is evident between the 35 two separate sections of coontail and reflects again the better growth exhibited by this plant. Table 6. Percent organic phosphorus and nitrogen in cropped mate- rial from period I. . First Crop % % N/P % Section g Dry Wt. N P Quotient Protein Stream A 1 47.3 2.67 0.61 4.4 16.7 2 4.1 1.80 0.43 4.2 11.2 3 32.5 1.25 0.39 3.2 7.8 4 20.4 2.06 0.75 2.8 12.9 5 13. 1 1.76 0.38 4.6 11.0 6 35.0 1.50 0.46 3.3 9.4 Stream B 1 11.9 1.2 0.3 4.0 7.5 2 6.8 1.3 0.3 4.5 8.3 3 26.9 0.6 0.2 4.5 4.0 4 14.2 0.9 0.2 4.2 5.6 5 4.7 1.4 0.3 4.5 8.5 6 9.1 1.0 0.2 5.2 6.2 The total mg of nutrients per section is, of course, di- rectly related to the production from that section. As a result, in some cases a section with a lower percentage of nutrients far exceeds 36 another in actual mg of nutrients. This was evident several times in the case of Vallisneria. Table 7. Percent organic phosphorus and nitrogen in cropped mate- rial from period II. . 2nd Crop % % N/ P % Sect1on g Dry Wt. N P Quotient Protein Stream A 1 47.6 3.98 0.80 5.0 25.0 2 2.9 1.80 0.40 4.4 11.2 3 27.6 1.80 0.39 4.7 11.4 4 52.5 3.30 0.66 5.0 20.6 5 4.1 2.30 0.39 5.8 14.0 6 29.0 1.70 0.37 4.6 10.5 Stream B 1 38.7 3.0 0.4 7.6 19.0 2 2.8 1.0 0.3 3.5 6.3 3 20.2 0.8 0.2 4.1 4.9 4 50.5 1.3 0.3 4.1 8.0 5 2.6 1.5 0.3 5.8 4.5 6 20.4 0.9 0.2 4.3 5.4 At the end of the first growth period, some of the plants were in poor condition. This was more pronounced in control stream B and, as previously mentioned, several of these sections were re- planted. In an effort to get the control plants in better condition, 37 nitrate and phOSphate enrichment (2 mg/l) was added to stream B for 50 hours. This treatment succeeded in aiding the plants of the con- trol. A result of this interim exposure to nutrients shows up in the total nitrogen and phosphorus content of the control plants in period II. These values are higher in contrast to corresponding values from period I and the differences in relation to the treated stream A are not as great. The N/P quotients remain fairly uniform for any one growth period in both stream A and B. These quotients which reflect the ratio of phOSphorus and nitrogen range slightly higher during the second period than the first. This coincides with the general in- creases in production between the two growth periods and could be directly related to the interim treatment of stream B. In terms of nutrient uptake per gram of plant material, there was an obvious gradient established between the three plants (Table 8). This could be due to the plants at the head of the channel being exposed to higher concentrations of incoming nutrients as a result of a nutrient gradient within each stream. The differences be- tween treated and non-treated plants were correspondingly greatest between the upper sections. Here again it must be noted that Cerato- phyllum had more production than the other plants. A sharp gradient in protein content was also noted between the different plants. This was a result of the differences in nitrogen 38 uptake by the plants. In the case of coontail, the results show that the protein content can be almost doubled by enrichment at the levels used in this study. The fact that the Ceratophyllum in stream A was 19% protein indicates a potential use for cropped macrophytes as a source of protein. Table 8. Average organic nutrients and percent protein for each macrophyte community. mg P mg N % g Dry Wt' mg P g DW mg N g DW Protein Stream A Ceratophyllum 167. 8 1176.0 7.1 4172 24. 8 18.8 Vallisneria 24.2 96. 7 4.1 440 18.5 11. 8 Elodea 124.6 505.6 4.0 1923 15.4 9.8 Stream B Ceratophyllum 115. 3 343.0 2. 9 2091 18.1 10. 0 Vallisneria 16.8 48.0 2.8 221 13. 1 8.2 Elodea 77.0 136.0 1.8 595 7.7 5.1 The total amount of phosphorus and nitrogen remaining in the system over the 30 day study period was calculated using the in- coming flow rate, the concentration of incoming nutrients and the 39 average nutrient reduction within the system. The resulting values of 33 g of phosphorus and 119 g of nitrogen were compared with the organic nutrient totals from analysis of cropped material, uncropped material, and bottom sediments. Only 29.6% of the phosphorus and 32. 6% of the nitrogen theoretically remaining in the system was accounted for in the totals of analyses. There are a number of explanations for such a large discrepancy. Undoubtedly there was some nutrient adsorption to the sides, bottom, and substrate of the channels and also some loss of nutrients by evaporation. However, the most likely source of such alarge discrepancy in balance lies in shortcoming of applied proce- dures. Tests for total phOSphorus and total nitrogen were not run on the daily discharge from the enriched stream. As an unfortunate consequence, no measurement was obtained for that portion of nutri- ents taken up by planktonic forms which washed out of the system. This could account for a large portion of the disparity in nutrient balance. Another area for which no tests were taken concerns the release of nutrients during the night periods of the study. Any nutri— ents released at night would have washed out of the system without being measured. SUMMARY 1. Macrophyte community production was measured in two artificial outdoor streams. The upstream-downstream nutrient uptake and removal relationships were studied in these streams. 2. Ceratophyllum demersum maintained the best condi— tion over the study period and had the least attached periphyton. Elodea canadensis and Vallisneria americana supported dense growths of mixed periphyton and did not grow as well. 3. Production measured by increase in dry weight mate- rial noticeably increased with phOSphate and nitrate enrichment. There was little difference in production between the 4 mg/l and 2 mg/l nutrient levels. 4. The Elodea canadensis community produced 5. 4 g dry wt/mzl day followed by the Ceratophyllum demersum community at 4. 9 g dry wt/ m2/ day and Vallisneria americana community at 0. 7 g dry wt/mz/day. A large portion of the production by the Elodea com- munity took place at the lower depths and was not recorded in crop- ping procedures. 5. The phosphate level was reduced by an average of 18. 7% and the nitrate level reduced by 19. 5% in the artificial streams. 40 41 It is likely that these values could be increased using longer retention periods. 6. There was a definite increase in the organic phospho- rus and nitrogen content of the cropped plant material in stream A. The N/P quotient for the plants was an average 4. 5 and remained rel- atively constant for all the plants. 7. The Ceratophyllum demersum contained the highest percentage of organic phOSphorus and nitrate and the highest average protein content of 19%. 8. The amount of detritus accumulated over a 30 day pe- riod was considerably higher in the enriched stream. The Elodea community deposited the greatest amount of detritus at a rate of 11. 3 g dry wt/mZ/day. LITERATURE CITED APHA, AWWA, WPCF, 1965. Standard Methods for the Examination of Water and Waste Water. 12th Edition. Boyd Printing Co., Inc., Albany, N.Y. 769pp. Bartsch, A. G., and M. O. Allum. 1957. An Oxidation Pond as a Tertiary Treatment Device. ASCE Sanitary Engineering Division. Vol. 91 SA 3:31-44. Beck, W. M. 1962. The Biological Removal of Nitrogenous Com- pounds from Sewage Treatment Plant Effluents. Semi- nar on Biological Problems in Water Pollution. Public Health Service Publication No. 999-WP-25z306-308. Bogan, R. H. 1960. The Use of Algae in Removing Nutrients from Domestic Sewage. Algae and Metropolitan Wastes, R. A. Taft Sanitary Engineering Center Technical Report W 61-3. Blackburn, R. D., J. M. Lawrence, and D. L. Davis. 1961. Effects of Light Intensity and Quality on the growth of Elodea densa. Weeds 92251—257. Hasler, Arthur D. , and Elizabeth Jones. 1949. Demonstration of the Antagonistic Action of Large Aquatic Plants on Algae and Rotifers. Ecol. 30:359-364. Hutchinson, G. E. 1957. A Treatise on Limnology. John Wiley and Sons, Inc. Vol. 1:1015pp. Kevern, Niles R. 1962. Primary Productivity and Energy Relation- ships in Artificial Streams. Thesis Ph. D. Michigan State University. Loehr, R. C., and R. L. Stephenson. 1965. An Oxidation Pond as a Tertiary Treatment Device. ASCE Sanitary Engineer- ing Division. Vol. 91 SA 3:31-44. 42 43 Mackenthun, K. M., W. M. Ingram, and R. Porges. 1964. Limno- logical Aspects of Recreational Lakes. U. S. Public Health Service Publication No. 1167. 176pp. Mackenthun, K. M. 1965. Nitrogen and PhOSphorus in Water. Pub— lic Health Service Publication 1305. 112pp. Merkle, M. G., and S. M. Fertig. 1963. Some Effects of Plant Growth on the Aquatic Environment. NEWCC Proceed- ings 17:432-438. Merkle, M. G., and S. N. Fertig. 1963. Problems and Techniques in Growing Certain Aquatic Species Under Greenhouse Conditions. NEWCC Proceedings 17:475-479. Odum, H. T. 1956. Primary Production in Flowing Waters. Lim- nology and Oceanography. 1:102-117. Penfound, W. T. 1956. Primary Production of Vascular Aquatic Plants. Limno. and Oceanogr. 1:92-101. Prescott, G. W. 1962. Algae of the Western Great Lakes Area. Wm. C. Brown Co., Inc., Dubuque, Iowa. 946pp. Rickett, H. W. 1922. A Quantitative Study of the Larger Aquatic Plants of Lake Mendota. Trans. Wisconsin Academy of Science. Vol. 20:501—527. Rickett, H. W. 1924. A Quantitative Study of the Larger Aquatic Plants of Green Lake, Wisconsin. Trans. Wis. Acad. Sci. Vol. 21:381-414. Ruttner, F. 1953. Fundamentals of Limnology. University of Toronto Press, Canada. 242pp. Sawyer, C. N. 1960. Chemistry for Sanitary Engineers. McGraw- Hill Book Co. , Ind. , New York. 367pp. Schuette, H. A. , and H. Alder. 1928. Notes on the Chemical Com- position of Some of the Larger Aquatic Plants of Lake Mendota. II Vallisneria and Potomogeton. Trans. Wis. Acad. Sci. Arts, Letters. Vol. 23:249-254. 44 Schulze, K. L. 1966. Biological Recovery of Waste Water. Jour. Water Pollution Control 38:1944. Stokes, R. M. 1960. The Effects of Limiting Concentrations of Nitrogen on Primary Production in an Artificial Stream. Thesis M. S. Michigan State University. Thomas, E. A. 1962. The Eutrophication of Lakes and Rivers, Cause and Prevention. Seminar on Biological Problems in Water Pollution. Public Health Service Publication No. 999-WP-25:306-308. Welch, P. S. 1948. Limnological Methods. McGraw—Hill Book Co., Inc., New York. 367pp. Wetzel, R. G. 1964. A Comparative Study of the Primary Produc- tivity of Higher Aquatic Plants, Periphyton, and Phyto— plankton in a Large, Shallow Lake. Int. Revue Ges. Hydrobiol. 49:1:1-61. Wilkinson, R. E. 1963. Effects of Light Intensity and Temperature on the Growth of Waterstargrass, Coontail, and Duck- weed. Weeds 11:287-290. 45 88.85 n 00 saw .888 n mo macaw u m mquHHpcoU Haunoficougcm n .O .m o.o mm mm H.o mm ¢.m m0 mp om mH m.o mm mm m.o Him mm m om Hum NH H.o mm mm mm om mm 00 mH. Hm HH «to cm mH. m.o H .m mH. m cm mm 3 H .o mm mm Nd- Him m.m m mp mu m m.o om m.m m6 om mm 00 mp om m 0.0 m.m mm To Hm N..m m mm mm H. 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Original starting plant mass and formulas for develop- ing uniform origin. A = Original Planted Material B = Material Replanted Before Second Period C = Positive Uncropped Growth or Negative Lost Material Formula Original for First Cropping F.O. = Ad: 1/4C Formula Original for Second Cropping Case 1. No Replant F.O. = A d: 3/4 C Case 2. Replant F.O. = B i 1/4C Formula Original for Total Cropped Material F.O. lst crop + F.O. 2nd crop F.O. = 2 Formula Original for Cropped Plus Uncropped Growth Case 1. No Replant and No Lost Material F. O. = A Case 2. No Replant and Lost Material F.O. = A - 1/2 C Case 3. Replant and No Lost Material F. O. = A + B Case 4. 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