1 AN ABSTRACT OF THE THESIS OF RONALD MAURICE EDDY for the MASTER OF SCIENCE (Name f Student) (Degree) in FISHERIES & WILDLIFE presented on 2-7 i7/ (Major Department) (Dafe) Title: THE INFLUENCE OF DISSOLVED OXYGEN CONCENTRATION AND TEMPERATURE ON THE SURVIVAL AND GROWTH OF CHINOOK $ALMO Abstract approved: MB Signature redacted for privacy. Dean Shumw Chinook salmon, Oncorhynchus tshawytscha, were reared from fertilization of the eggs to several weeks after complete yolk absorption atdissolvedoxygenconcentrationof3.5, 5.0, and7.3 mg/literandair-saturation at temperatures of l5, 12.0, 13.5, and 15.0 C. Decrease of either temperature or dissolved oxygen concentration resulted in increase in the median (5 0%) hatching time of fry. Increase of temperature or decrease of dissolved oxygen concentration decreased mean dry weights of newly hatched fry and survival. Temperatures above C resulted in substantially reduced survival at all dissolved oxygen levels tested. Large increases in mortality occurred during late embryonic and sac fry stages, the amount of mortality during each period being directly related to the prevailing test conditions.
2 The Influence of Dissolved Oxygen Concentration and Temper8ture on the Survival and Growth of Chinook Salmon Embryos and Fry by RONALD MAURICE EDDY A THESIS submitted to OREGON STATE UNIVERSITY in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE June 1972
3 APPROVED: (Sinature redacted for privacy. i5 I, Associate Professor of Fisheries in charge o major Signature redacted for privacy. 1ad of Department of Fisheries and *ildlife Dean of Graduate School Signature redacted for privacy. Date thesis is presented 27 5:'7%c/4 (7/ Typed by Mary Lee Olson for RONALD MAURICE EDDY
4 ACKNOWLEDGEMENTS Sincerest appreciation is extended to Mr. Dean L. Shurnway, Associate Professor of Fisheries, Oregon State University, for his guidance and assistance throughout the course of this investigation. Thanks are due Dr. John R. Donaldson, Associate Professor of Fisheries, for his assistance in reviewing this manuscript. The assistance of the staff and the many students who worked at the Oak Creek Fisheries Laboratory, Oregon State University, during the course of my study is gratefully acknowledged. Special appreciation is extended to Mr. Lowell Moore for his assistance in collecting data and maintaining the experimental apparatus.
5 TABLE OF CONTENTS Page INTRODUCTION V 1 EXPERIMENTAL MATERIAL, APPARATUS, AND METHODS 5 Experimental materials 5 Experimental apparatus 5 Experimental methods 10 RESULTS 14 DISCUSSION 35 LITERATURE CITED 44
6 Figure LIST OF FIGURES Page 1 Flow diagram of the experimental apparatus 6 2 Cross-sectional view of a test chamber 9 3 The relation between the median hatching time of chinook salmon and the dissolved oxygen concentrations at each temperature tested 17 4 The median hatching time of chinook salmon in relation to the temperatures at each dissolved oxygen concentration tested 19 5 A three-dimensional diagram showing the median hatching time of chinook salmon in experiment 1 in relation to both dissolved oxygen concentration and temperature 21 6 A three-dimensional diagram showing the median hatching time of chinook salmon in experiment 2 in relation to both dissolved oxygen concentration and temperature 22 The relation between the mean dry weights of newly hatched chinook salmon fry and the dissolved oxygen concentrations at each test temperature 23 8 The relation between the mean dry weights of newly hatched chinook salmon fry and the temperatures at each dissolved oxygen concentration tested 24 9 The per:cent survival of chinook salmon from time of fertilization to termination of experiment The percent survival of chinook salmon from time of fertilization to termination of experiment 2 27
7 Figure Page 11 The relation between the percent survival of chinook salmon at termination of the experiments and the dissolved oxygen concentrations at each temperature tested The percent survival of chinook salmon at termination of the experiments in relation to the temperatures at each dissolved oxygen concentration tested A three-dimensional diagram showing the percent survival of chinook salmon at termination of experiment 1 in relation to both temperature and dissolved oxygen concentration A three-dimensional diagram showing the interrelation between percent survival of chinook salmon at termination of experiment 2, dissolved oxygen concentration, and temperature 34
8 LIST OF TABLES Table Page Means and ranges of water temperature and dissolved oxygen concentration, median hatching time of fry, mean dry weight of newly-hatched fry, and percent survival of chinook salmon. 15
9 THE INFLUENCE OF DISSOLVED OXYGEN CONCENTRATION AND TEMPERATURE ON THE SURVIVAL AND GROWTH OF CHINOOK SALMON EMBRYOS AND FRY INTRODUCTION The survival, development, and growth of chinook salmon, Oncorhynchus tshawytscha, embryos and fry under natural conditions depends to a large degree on the temperature and dissolved oxygen concentration of the water. During the time of spawning, elevated water temperatures resulting from factors such as irrigation return water, warm water releases from dams, and high seasonal temperatures may result in conditions unfavorable for survival and growth. Survival of embryos and fry exposed to only moderately high temperatures may be greatly decreased with reduction of dissolved oxygen concentration. Factors such as introduction of putricible substances into the water and siltation due to various agricultural and forestry practices may result in dissolved oxygen concentrations of intragravel water far below the air-saturation level. Even at sublethal levels, temperature and dissolved oxygen concentration may alter the time of hatching and the size of the newly hatched fry, Extensive research has been conducted on the influence of dissolved oxygen concentration on the survival, growth and delay in hatching of salmonid embryos reared at nearly constant temperatures. Good survival of embryos of various salmonid fishes, rainbow and
10 steelhead trout, Salmo gairdneri, coho salmon, Oncorhynchus kisutch7 chinook salmon, and sockeye salmon, Oncorhynchus nerka, reared under laboratory conditions has often been reported when the eggs were exposed continuously, from time of fertilization to hatching, to mean dissolved oxygen concentrations as low as 2. 1 to 3. 0 mgi liter at temperatures of 8 C to 11 C (Hamdorf, 1961; Silver, Warren and Doudoroff, 1963; Shumway, Warren,and Doudoroff, 1964; Brannon, 1965; Chapman, 1969). Silver et al. (1963) reported 100% mortality of chinook salmon embryos reared from time of fertilization to hatching at 1.6 mg/liter of dissolved oxygen at 11 C. Mean weights (wet and dry) or volumes of salmonid alevins (sac fry) at time of hatching at dissolved oxygen levels of 2. 5 to 3. 0 mg/liter have been reported to be approximately one-fourth to one-half those of controls reared at near air-saturation levels (Mason, 1969; Shumwayetal., 1964; Chapman, 1969; Silver etal.., 1963; Hamdorf, 1961). With reduction of dissolved oxygen concentration the time required to reach hatching by embryos of coho and chinook salmon, and rainbow and steelhead trout has been reported to increase (Shumway et al., 1964; Chapman, 1969;Hamdorf, 1961; Silver etal., 1963). Garside (1959, 1966) reported that embryos of lake trout, Salvelinus namaycush; brook trout, Salvelinus fontinalis, and rainbow trout exposed, from time of fertilization to hatching, to different dissolved oxygen concentrations at different temperatures required more time to reach various 2
11 developmental stages with decrease of either temperature or dissolved oxygen concentration. Although smaller at time of hatching, fry reared at reduced dissolved oxygen levels may attain nearly the same maximum size upon completion of yolk absorption as fry reared at more optimal concentrations (Chapman, 1969; Doudoroff and Shumway, 1967, 1970; Brannon, 1965). Doudoroff and Shumway (1970) concluded, after an extensive review of the pertinent literature on the influence of dissolved oxygen on larval growth, that the efficiency of conversion of yolk to body tissue by salmonid alevins is not materially impaired by reductions of dissolved oxygen concentration to 5 mg/liter when other conditions are favorable. The time required by alevins to complete yolk absorption, however, progressively increased with decreasing dissolved oxygen concentration. Other researchers have studied the effects of temperatures on the survival and growth of salmonid embryos and fry reared at near air-saturation levels of dissolved oxygen, Combs and Burrows (1957) reared chinook salmonembryos fromtime of fertilization to hatching at temperatures ranging from 1. 7 C to C Very high mortality was observed at the lower temperatures (1. 7 C and 3. 1 C); only slight mortality was reported at intermediate test temperatures (4. 4 C to 14. 2C). With increase in temperature from C to C, however, mortality significantly increased. Data 3
12 4 presented by Olsen and Nakatani (1968) showed that the mortality of chinook salmon, reared from time of fertilization until well after complete yolk absorption at seven different seasonally fluctuating temperatures regimes, was markedly increased when the embryos were exposed to temperatures of C and above at the start of the incubating period. Donaldson (1955) reported that any increase in the exposure time of chinook salmon embryos to temperatures of, 17. 4, 18. 3, and 19.4 C resulted in increased mortality. Excessive mortality of chinook salmon resulting from exposure to the elevated temperatures, hbwevèr, was not evident in many instaices until the latter part of the sac fry stage. The laboratory investigation reported here was conducted to determine the influence of dissolved oxygen concentration and temperature on chinook salmon embryos and fry reared from time of fertilization to several weeks after completion of yolk absorption. Determinations were made of time to median (50%) hatch, weight of fry at hatching, and survival. Two experiments were conducted simultaneously from October 1970 through February 1971 at the Oak Creek Fisheries Laboratory of the Department of Fisheries and Wildlife, Oregon State University, Corvallis, Oregon.
13 5 EXPERIMENTAL MATERIALS, APPARATUS, AND METHODS Experimental Materials The chinook salmon eggs used in this investigation were obtained from the Fish Commission of Oregons Bonneville Fish Hatchery located on the Columbia River. Eggs were taken from two females, a separate female for each experiment, and both batches of eggs fertilized with sperm from the same three males. The mean diameters of the eggs used in experiments 1 and 2 were 8 mm and 7 mm, respectively. The eggs were allowed to water harden for one-half hr before transport to the Oak Creek Fisheries Laboratory. Water temperature at the hatchery was 8 C at the time the eggs were fertilized. A 2 C rise in temperature occurred during the 3-hr transportation period and an additional rise of 3. 5 C occurred during the ensuing 2-hr period required to distribute the eggs in the experimental apparatus. Experimental Apparatus The experimental apparatus used in this study was designed for exposing chinook salmon embryos and fry to flowing water of controlled dissolved oxygen concentration, temperature, and velocity (F i g. 1). Four, :160-liter wooden water baths were constructed, each containing four pairs of test chambers. All test chambers within a water bath were maintained at one of four test temperature s
14 Plastic Float Valve Return Water Thermoregulator Glass Column Test Chamber Immersion Heater Overflow \0 Flowmeter r 2-Stage Reduction Valve Gas Dispersion Tube Crosssupport Stopcock 1 Gas Manifold Pyrex Tubing Stopcock Submersible Pump Gas Flowxneter. S -r Needle Valve Figure 1. Flow diagram of the experimental apparatus (two of eight test chambers located in one of four water baths). Successive stages of the water flow apparatus are connected with vinyl tubing.
15 7 with each pair of test chambers maintained at one of four dissolved oxygen concentrations. Filtered water from a small, spring-fed stream was supplied through polyethylene pipe to the constant-temperature room housing the experimental apparatus. The incoming water flowed into four separate headboxes via plastic float valves, The water in each headbox was aerated with compressed air and brought to the desired temperature by a thermostatically controlled, stainless - steel immersion heater. The heated water from each headbox then flowed into four vertical 50-mm 0. D., 75-cm high glass columns. The water was introduced near the top of each column through a 9-mm diameter glass tube inserted into the column through the stoppered bottom. Flows of nitrogen and oxygen to the glass columns were regulated by 2-stage reduction valves and gas flowmeters and were introduced at the bottoms of the columns through fritted-disc gas dispersion tubes inserted through the stoppered bottoms. By withdrawing water from the bottoms of the columns, count erfiows of gas bubbles and water were obtained. Each of the 16 glass columns supplied water to two test chambers. A flow-adjustment stopcock and a calibrated ball-displacement flowmeter were used to reg4late the flow of water to each test chamber. Water was introduced at the bottom of the test chamber
16 at a rate of approximately 400 mi/mm. The velocity of the water passing the eggs in the test chamber was calculated to be approximately 100 cm/hr. The water then flowed up through the test chamber and was discharged into the water bath through a row of small holes around the top of the test chamber (Fig. 2). The test chambers were constructed from two-piece, polypropylene Buchner funnels (Fig. 2). The bottom section of the test chamber was firmly seated in a wooden cross-support. The top section of the test chamber was a 18-cm diameter, 15-cm high cylinder with a perforated bottom and an open top. A single layer of 4-mm glass beads was placed on the perforated bottom to help evenly distribute the flow of incoming water. A small diameter glass tube inserted into thetest chamber through a rubber stopper was used to remove water samples for dissolved oxygen determinations (Fig. 2). The top section of the test chamber was initially covered with a piece of perforated sheet metal. When the sac fry in a test chamber neared complete yolk absorption, the sheet metal cover was replaced with a piece-of small-mesh screen to allow more light to enter the test chamber. The constant temperature room housing the experimental apparatus remained unlit, aside from daily check periods, until the most rapidly developing sac fry neared complete yolk absorption. 8
17 9 DISCHARGE COVER / PORTS / \/ O 0 EGGS GLASS BEADS PERFORAT ED PLATE WATER SAMPLING TtJBF PYREX TUBING TYGON TUBING WATER Figure 2. Cross-sectional view of 1 of 32 test chambers used to rear chinook salmon embryos and fry.
18 At this time the room was artificially illuminated for 12 hrs each day until termination of the experiments. In order to conserve heated water, approximately 60% of the water discharged from the test chambers into a water bath was returned to the headbox by a small submersible pump located in th water bath (Fig, 1), The return flow of water was monitored and. regulated by a flow-adjustment stopcock and flowmeter. The submersible pump also served to circulate water within the water bath to help maintain a uniform temperature throughout. The remaining 40% of the water entering the water bath was discharged to a floor drain via an overflow tube. 10 Experimental Methods Renewal and recirculation flows of water were started and adjusted several days prior to the start of the experiments. Duriug this time, the dissolved oxygen concentrations and temperatures of the water were gradually brought to the desired levels. Checks of the experimental apparatus were made twice daily, once between the hours of 8:00 and 10:00 A. M. and again betweenthe hours of 800andl0:0O P.M. Gas andwaterflowmeterreadings, water temperatuxs, and dissolved oxygen concentratiors were recorded during each check period, and adjustments made when necessary. Dissolved oxygen concentrations re determined using the azide modification of the
19 iodometric method (American Public Health Association et al.,, 1965). Dissolved oxygen concentrations were determined and recorded to the nearest mg/liter. Temperatures were recorded to the nearest 0.05 degre centigrade. At the start of the experiments, approximately 250 eggs ( ) were placed in a single layer on the glass beads in each test chamber. As previously stated, each glass column supplied water to two test chambers. By placing the eggs from one female into one test chamber in each pair serviced by a column, and the eggs from the second female in the other test chamber, eggs from both females we-re exposed to identical conditions of temperatures and dissolved oxygen concentrations throughout the test period. Dead eggs were not removed from a test chamber before the eyed stage had been reached by the live embryos in that test chamber.. Thereafter, dea.d eggs and embryos were removed and recorded during the first check period each day. All dead eggs were cleared in Stockardts solution (Rugh, 1952) in order to determine whether development had begun. Eggs were termed non-viable when, after clearing in Stockard's solution, embryonic tissue could not be observed. Embryos and fry held in the test chambers periodically sampled for wet and dry weight determinations. The first sample of approximately 30 embryos was removed from a test chamber when the embryos in the test chamber had reached the eyed stage of 11
20 development. Once a sample of eggs was obtained, the chorion was removed from each embryo in the sample. The chorion was pierced at an oblique angle with a sharp needle, and micro-tweezers used to peel back the chorion, exposing the embryo. After removal of the chorions, the embryos were divided into two subsamples of nearly equal numbers and killed in a solution of MS 222. The embryos, with yolk intact, in one subsample were blotted with a damp towel and weighed in aggregate.. The embryos of the remaining subsample were handled in the same manner, except that the yolk material from each embryo was excised prior to weighing. After wet weight determinations, both subsamples of embryos were placed in a drying oven and held at approximately 70 C for 5 days. The subsamples were then removed from the oven, placed. in a desiccator to cool, and dry weights determined. The amount of yolk material present could then be determined by subtracting the mean weight of the embryos without yolk material from the mean weight of the embryos with yolk material When 95% to 100% of the embryos in a test chamber had completed hatching, a second sample of approximately 30 newly hatched fry was taken. The fry were divided into two subsamples of nearly equal numbers and, aside from chorion removal, handled in the same manner described earlier. Periodic sampling of the 12
21 fry remaining in the test chambers was continued until 30 days after the fry were started on feed. Feed (chopped, live tubificid worms) was provided approximately 10 days prior to complete yolk absorption. Fry were fed to excess twice daily. 13
22 14 RESULTS The temperature and dissolved oxygen concentration of wat-er flowing past developing chinook salmon embryos and fry were found to the- markedly influence the length of time required to reach hatching, size at time of hatching, and survival. The means and ranges of temperatures and dissolved oxygen concentrations to which embryos and fry were subjected during this study are presented in Table L The time in days from fertilization of the eggs to median hatching, the mean dry weight of fry at time of near-complete hatching, and the percent survival at termination of the experiments are also given in Table 1. The near air-saturation levels of dissolved oxygen varied from 11.0 mg/liter at C to mg/liter at C (Table 1). Somewhat less variation in mean dissolved oxygen concentration occurred at the three lower levels tested. In order to facilitate the presentation of data in some of the figures, similar mean dissolved oxygen levels from the four test temperatures were averaged and the values obtained (lo.5r 7.3, 5.O,and3.5 mg/liter) usedin aphing the data In other figures, the mean values obtained at each temperature were used in graphing the data (Table 1). The curves presented in Fig. 3 relate the median hatching time of the embryos, in days after fertilization, to dissolved oxygen
23 TABLE 1: Means and ranges of water temperature and dissolved oxygen concentration, median hatching time of fry, mean dry weight of newlyhatched fry, and percent survival of chinook salmon in experiments 1 and 2. Experiment No. 1 2 Mean temperature and range (C) ( ) ( ) ( ) ( ) ( ) ( ) ( ) Dissolved oxygen (irg/1it4r) Mean Range , i Median hatching time-!! Mean dry weight of fry 1 (inz) Sampling day Percen4' Survival Davs' 66 66* 66* 66* * 66* 66* *
24 TABLE 1: (Continued) Mean temperature Dissolved oxygen Median Mean dry Experiment and range.(mz/liter) hatching weight of fryv Sampling Survival No. (C) Mean Range timej (mg) day/ Percents' Days ( ) S Median (50%) hatching time in days after fertilization. 2 Mean dry weight of newly-hatched fry with yolk removed taken at time of near-complete (95-100%) hatching. 3 Time, indays after fertilization, at which the near-complete hatching sample was taken from each test chamber. 4 Only viable eggs were used in calculating percent survival. 5 Time, in days after fertilization, at which percent survival was calculated. Asterisk indicates premature termination due to equipment failure.
25 17 Experiment 1 _ >. 30 z 60 Experiment C DISSOLVED OXYGEN CONCENTRATION IN MILLIGRAMS PER LITER Figure 3. The relation between the median hatching time of chinook salmon and the dissolved oxygen concentrations at each temperature tested.
26 concentrations at each test temperature. Decrease of dissolved oxygen concentration resulted in an increase in the number of days required by the embryos to reach median hatching at all temperatures tested. In experiment 1, decrease of dissolved oxygen concentration from near air-saturation to 3.5 mg/liter at 10.5, 12.0, 13.5, and C resulted in an increase in median hatching time of 6, 8, 11, and 16%, respectively. In experiment 2, similar decrease of dissolved oxygen concentration at the same temperatures resulted in increases in median hatching time of 7, 8, 10, and 9%. Figure 4 shows the relation between the median hatching time of chinook salmon fry and temperatures at averaged dissolved oxygen concentrations of 3. 5, 5. 0, 7. 3, and mg/liter. At all dissolve& oxygen levels tested, median hatching time decreased with increasing temperature. Increase in temperature from C to C and. from 12.0 C to 13.5 C resulted in similar decreases in the median hatching time. Slightly smaller decreases in median hatching time occurred with further increase of temperature from 13.5 C to l5.0 C. Increase in temperature from 10.5 C to 15.0 C exhibited a far greater influence on median hatching time (25% to 34% decrease, experiment 1; 30% to 32% decrease, experiment 2) than did decrease of dissolved oxygen concentration from near air-saturation to 3.5 mg/liter (6% to 16% increase, experiment 1; 7% to 10% increase, experiment 2). 18
27 19 Experiment MG/L V 5.OMG/L L\ 7.3MG/L MG/L Experiment 2 TEMPERATURE (C) Figure 4. The median hatching time of chinook salmon in relation to the temperatures at each dissolved oxygen concentration tested. Dissolved oxygen concentration are averages of similar dissolved oxygen levels from each test temperature.
28 20 The relation of median hatching time of chinook salmon fry to both dissolved oxygen concentration and temperature is represented for the experiments by separate three-dimensional graphs in Fig. 5 and 6. As can be seen from the graphs, reductions of dissolved oxygen concentration below the air-saturation level resulted in increases in. the number of days required by the embryos to reach median hatching. Increase in temperature from C to C resulted in large decreases in the time required by the embryos to reach median hatching. Figure 7 relates the mean dry weights of newly hatched fry to the dissolved oxygen concentrations at temperatures of 10.5, 12. O, 13. 5, and C. In general, mean dry weights of fry in both experiments decreased with reduction of dissolved oxygen concentrátion. In experiment 2 at C, however, the size of fry was reduced at the lowest dissolved oxygen level tested only. This relationship was not observed at C in experiment 1 as the size of fry decreased with any reduction of dissolved oxygen concentration. The relation between the mean dry weights of newly hatched fry and temperatures at averaged dissolved oxygen concentrations of about 3. 5, 5. 0, 7. 3, and mg/liter is presented in Fig. 8r In general, increase in temperature resulted in decrease of the size of the fry at time of hatching. The reasons for the unpredictable variations in the mean dry weights of fry are unknown.
29 60 z IJISSOL VEb 8 IJ\J IGRAMS Figure 5. A three-dimensional diagram showing the median hatching time (in days after fertilization) of chinook salmon in experiment 1 in relation to both oxygen concentration and temperature.
30 60 z z :iz z S 8 bissg: 9 LIT Figure 6. A three-dimensional diagram showing the median hatching time (in days after fertilization) of chinook salmon in experiment 2 in relation to both oxygen concentration and temperature.
31 23 Experiment Experiment DISSOLVED OXYGEN CONCENTRATION IN MILLIGRAMS PER LITER Figure 7. The relation between the mean dry weights of newly hatched chinook salmon fry and the dissolved oxygen concentrations at each test temperature.
32 24 Experiment MG/L V MG/LL MG/L MG/L Experiment TEMPERATURE (C) Figure 8. The relation between the mean dry weights of newly hatched chinook salmon and the temperatures at each dissolved oxygen concentration tested. Dissolved oxygen concentrations are averages of similar mean dissolved oxygen levels from each test temperature.
33 The curves presented in Fig. 9 and 10 show the changes in per cent survival of chinook salmon embryos and fry from time of fertilization to termination of the experiments at the various temperatures and dissolved oxygen concentrations tested. Percent survival was calculated for the embryos and fry in each test chamber prior to the removal of samples. percent survival. Percent survival was calculated using the following formula: where A B Only viable eggs were used in calculating (A-B) A x C=D number of fish present in the test chamber at the start of a sampling period, mortality incurred during the sampling period, C = percent survival at the end of previous sampling period D (C = 100% for the first sampling period from each test chamber), and percent survival at the end of the sampling period in question. The calculated percent survival of embryos and fry, for each sampling period from each test chamber, was then plotted against time and curves fitted by eye to the points. Survival curves for chinook salmon reared at reduced oxygen levels at C are not presented in Fig. 9 and 10. A partial failure of the water system 66 days after the start of the tests 25
34 C too Figure 9. DAYS AFTER FERTILIZATION The percent survival of chinook salmon from time of fertilization to termination of experiment 1. Dissolved oxygen concentrations are averages of similar mean dissolved oxygen levels from each test temperature. Median (50%) hatching and start of feeding are designated by 0 and, respectively.
35 C C 0 -' Figure DAYS AFTER FERTILIZATION The percent survival of chinook salmon from time of fertilization to termination of experiment 2. Dissolved oxygen concentrations are averages of similar mean dissolved oxygen levels from each test temperature. Median (50%) hatching and start of feeding are designated by 0 and, respectively.
36 resulted in discontinuation of water flows to all test chambers at that temperature, with excessive mortality occurring at the reduced dis-: solved oxygen levels. Mortality continued at the reduced dissolved oxygen vels for approximately 3 weeks after water stoppage. Due to the inability of distinquishing between mortalities resulting from stress incurred during failure of the water system and mortalities resulting from the test conditions, accurate estimates of mortality resulting from the test conditions only could not be made and for this reason the survival curves were not presented. Reported percent survivals of chinook salmon reared at C at all dissolved oxygen levels tested were calculated 66 days after fertilization, the time of failure of the water system. The percentages reported probably would have been slightly lower if the fry had been reared through the entire, relatively critical, sac fry stage of development. Major reductions of survival of chinook salmon sac fry occurred at nearly all test conditions (Fig. 9 and 10). Increase of temperature or decrease of dissolved oxygen concentration increased mortality occurring during this time. Examination of data on the rate of yolk utilization, obtained during the study but not presented here, revealed that the onset of the periods of major reduction of survival occurred progressively earlier with either increase of temperature or decrease of dissolved oxygen concentration. 28
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Western Blotting Sive Lab Protocol March 2007 Prepare samples: For zebrafish embryos: Option 1: Take live embryos and put into 1.5 ml tube with E3. Centrifuge gently for 1-2 minutes -yolk lipids will rise
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Example: 2 Design low rate trickling filter for secondary treatment of sewage generated from 10000 persons with rate of water supply 170 LPCD. The BOD 5 after primary treatment is 110 mg/l and BOD 5 of
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