THE EFFECT OF TURBIDITY CAUSED BY SEWAGE POLLUTION ON THE PRODUCTIVITIES OF IRIDAEA FLACCTDA, PHYLLOSPADIX TORREYI, AND LAMINARIA SETCHELLII John Hainsworth Hopkins Marine Station Pacific Grove, Calif. June 2, 1970 INTRODUCTION Observation of the floral community surrounding an intertidal sewage outfall shows that some species of benthic algae and marine plants are able to exist in areas of high sewage concentrations, while other species cannot tolerate this altered environment. However, even the most resistant species are small and stunted in the immediate vicinity of the outfall. Since one of the most salient character¬ istics of sewage pollution is the increased turbidity caused by the suspended particulate matter in the sewage, it is logical to hypothesize that the increased turbidity effects the viability of the marine flora by reducing the amount of light received, therefore interfering with the normal photosynthetic process. The specific absorbtion spectrum of the sewage may also be a factor in determining the different tolerances of different species, since each has a unique pigment composition An experiment was designed to answer two questions: 1) does turbidity effect the productivity of marine flora, and 2) does turbidity have a different effect on different species. In all species studied the productivity decreased markedly at high turbidities. The productivities of the three species studied also showed differences in their rates of decline as the turbidity increased. 33. METHODS AND MATERIALS Two species of benthic algae (Iridaea flaccida and Laminaria setchellii) and one marine plant (Phyllospadix torreyi) were tested. These particular species were chosen for two reasons: 1) Iridaea flaccida and Laminaria setchellii are representative of the red and brown classes of benthic algae, while the pigments in Phyllospadix torreyi are sim¬ ilar to those in the chlorophytes, and 2) all three species are very common along the Monterey coast, where the study was conducted. Samples were collected from an unpolluted environment, cut into pieces weighing approximately 150 mg (dry weight), and put separately into 300 ml BOD bottles. (For both Iridaea flaccida and Laminaria setchellii, all pieces used for one test were cut from a single leaf.) The BOD bottles were filled with sea water which had a depressed dissolved oxygen content achieved by bubbling with nit¬ rogen for 8-10 minutes. The bottles were then placed on their sides in several two-gallon tubs containing water of different turbidities, and incubated at 12° C. A light of constant intensity was passed through a 3.5 cm. layer of turbid water to the test bottles. The bottles were submerged in water of three different turbidities, and in sea water, which served as a control. Sewage collected 1.2 meters from the Pacific Grove outfall was used in the most turbid bath, the other two baths being comprised of 1:2 and 1:4 dilutions of this sewage. Absorbtion 334 curves were determined for samples of sewage collected at various times of the day, and although the absorbances at specific wavelengths varied, the absorbtion spectra were parallel for all samples tested. A typical absorbtion curve for sewage is illustrated in Figure 1. Since the peak absorbtion is at 430 millimicrons, this wavelength was arbitrarily chosen as a measurement of turbidity. The turbidity of each bath was measured daily and adjusted by adding newly collected sewage. (The turbidity tended to drop after standing for longer than 24 hours.) The sewage was changed weekly to prevent gross changes in the ab¬ sorbtion spectrum due to bacterial growth or changes in particle size. Both Phyllospadix torreyi and Iridaea flaccida were submerged in the baths for one hour. Laminaria setchellii was incubated for two hours because of its slower rate of photosynthesis. A single test included duplicate bot¬ tles at each turbidity (all with one type of test organism), along with duplicate bottles in the control bath. When the test period ended, the bottles were removed from the baths and the dissolved oxygen in the water was measured using the Winkler method as described by Strickland and Parsons (1968). The dissolved oxygen in the water before the test was subtracted from the dissolved oxygen in each of the bot- tles, each piece of algae was dried and weighed, and the net productivity (in ml oxygen/mg dry weight/hour) was calculated using the formula: LB - IP net productivity - mg dry wt. hrs. where LB = dissolved oxygen in the light bottle, and IB dissolved oxygen in the initial bottle. After several trial runs, the test was repeated ten times with each species, and the first two runs for each species were discarded. RESULTS Figures 2, 3, and 4 represent a compilation of eight runs for each species tested. Duplicate bottles for each test at each turbidity were averaged, so that the three final graphs consist of eight points at each turbidity. Using these eight points, the means and standard deviations were calculated. The graphs show perecent of control product¬ ivity versus turbidity rather than net productivity versus turbidity because the control productivity in a single species varied from test to test since different plants were used. This was especially true for Iridaea flaccida. DISCUSSION Figures 2, 3, and 4 clearly indicate that the product- ivities of all three species studied are effected by tur bidity. Although the standard deviations (especially in the case of Laminaria setchellii) are quite large, it ap pears that the three different species respond differently 340 to increasing turbidity. Although the curves for Laminaria setchellii and Phyllospadix torreyi are quite similar, they are both very different from the Iridaea flaccida curve. While the productivities of both Laminaria setchellii and Phyllospadix torreyi drop in the 1:2 and 1:4 dilutions of the sewage, the Iridaea flaccida productivity remains ap proximately constant. The standard deviations which occur at some of the points in the graphs of Iridaea flaccida and Phyllospadix torreyi are misleadingly large, since one or two highly deviant values had a greater effect on the standard dev¬ iations, because there were only eight values determining the means. With many more tests using these two organisms the standard deviations would probably become much smaller. The values for Laminaria setchellii ranged quite widely and showed no indication that additional tests would sig nificantly lower the standard deviations. A number of features probably contributed to the undesirability of Laminaria setchellii as a test organism. Most important, Laminaria setchellii contains a sticky sap, which oozes out when the leaf is cut and then traps air bubbles when the bottle is filled. Although the bottles were filled immed- iately after the algae was cut, some air was probably still trapped. In addition, the very slow productivity rate of Laminaria setchellii (approximately eight times slower than that of Phyllospadix torreyi) maximized the errors caused by the trapped air bubbles. In spite of the wide variation in the results, Laminaria setchellii shows a distinct de¬ crease in productivity when the turbidity increases. The experimental results obtained for the three spe¬ cies studied correlate well with the locations of these three types in the vicinity of the Pacific Grove outfall at Point Pinos. None of the species is found within a 70meter radius of the outfall, where the turbidity at low tide ranges from .800 to 1.40 (turbidity as absorbtion at 430 millimicrons). Iridaea flaccida exists at one loca¬ tion with a turbidity of approximately .500 (the exact turbidity could not be measured since the rock was 7 meters offshore), and fairly commonly in areas where the tur¬ bidity was .100-.200. Neither Phyllospadix torreyi nor Laminaria setchellii appear except at low (.100-.200) turbidities, and all of these sites are washed from one side by clear water. A comparison of the absorbtion curve of sewage with the absorbtion curves of various plant pigments present in marine flora provides possible explanations for the detrimental effects of sewage turbidity on these organisms in general, and also for its different effects on the three species studied. All three types of chlorophylls, and most carotenoids have absorbtion peaks at 430-460 milli- microns (Franck, 1949), which is the range of maximum ab¬ sorbtion by the sewage. Since light at this wavelength is being absorbed by the sewage, the light available for ab¬ sorbtion by the chlorophyll is reduced. This predicted drop in productivity is well supported in Figures 2, 3, and 4. Although the absorbtion spectra for red, green, and brown algae are approximately the same, with absorbtion peaks at 430 and 660 millimicrons, Haxo and Blinks (1950) found that the action spectra of the red algae is greatly different than the action spectra of either green or brown algae. The action spectra of both green and brown algae approximate the absorbtion spectra, and have peaks at 430 and 660 millimicrons. (Phyllospadix torreyi, which is a marine flowering plant, contains the same pigments as does a green alga.) However, in red algae, the water-soluble phycobilin pigments are much more efficient at absorbing light than are the chlorophylls; because of this the red algae utilize light of 570 millimicrons most effectively, while very little of the light with wavelengths of 430-450 millimicrons is absorbed. The absorbtion spectrum of sew¬ age shows that approximately 30% more light of wavelength 570 millimicrons passes through than does light of 430 millimicrons. Red algae can be predicted, because of this fact, to be less effected by sewage turbidity than either green or brown algae. This prediction is supported by the results obtained in this investigation, using a single species characteristic of each algal type. Iridaea flaccida 37 is able to tolerate turbidity (up to a certain level) much better than either Phyllospadix torrevi or Laminaria set chellii. The ability to live in turbid water caused by sewage pollution is one reason that Iridaea flaccida, and perhaps red algae in general, can exist in water containing high sewage concentrations. However, turbidity is not the only parameter involved. Many other components of sewage, or properties of the different algal species themselves, also help to determine resistance to sewage pollution. Roy (personal communication) has found that several species of red algae in the vicinity of the Pacific Grove sewage out fall contain greater quantities of pigments (both chloro¬ phylls and carotenoids) than do the same types growing in unpolluted areas. This increased pigment concentration could possibly be a response to a decreased amount of light reaching the plant, due to sewage turbidity. Neither green nor brown algae have demonstrated this property. Schreiber (personal communication) has found that the productivity of several types of algae is decreased by increased chlorine concentrations in the water. Other pollutants found in sewage may also effect productivity, and these could be determinants of floral distribution in an outfall area. SUMMARY The productivities of all three species studied were found to decrease with increasing turbidity; this decrease 34 was as great as 35% in turbidities approximating the tur- bidity around the Pacific Grove sewage outfall. The ef¬ fects of this turbidity were also shown to differ for the three species studied. Iridaea flaccida exhibits a greater tolerance to sewage turbidity (except at very high values) than do either Phyllospadix torrevi or Laminaria setchellii. This finding is consistent with the prediction made for the effects of sewage turbidity on red versus green or brown algae using the action spectra of the different types. 4 ACKNOWLEDGEMENTS I wish to thank Dr. Ellsworth Wheeler for his assistance both in my research and in the writing of this paper. This research was supported in part by the NSF Undergraduate Research Program Grant +GY7288. REFERENCES CITED Franck, J. 1949. Photosynthesis in plants. Iowa State College Press. 450 pp. 1950. Photosynthetic action Haxo, F.L. and Blinks, L.R. spectra of marine algae Journal of General Physiology, 33 (4): 389-421. Strickland, J.D.H. and Parsons, T.R. 1968. A practical handbook of seawater analysis. Fisheries Research Board of Canada. 300 pp. Figure 1. Figure 2. Figure 3. Figure 4. FIGURE CAPTIONS Absorbance spectrum of sewage collected at the mouth of the Pacific Grove outfall. Change in productivity of Iridaea flaccida, as a function of increasing turbidity. Change in productivity of Phyllospadix torreyi, as a function of increasing turbidity. Change in productivity of Laminaria setchellii, as a function of increasing turbidity. ( O JONVadosav 0 10 E 3 0. 120 100 6 40 078 IRIDAEA FLACCIDA 720 415 175 TURBIDITY (Absorbance at 430 millimicrons 100 20 078 PHYLLOSPADIX TORREY 415 720 URBIDITY (Absorbance at 430 millimicrons) 1175 6( 40 078 LAMINARIA SETCHELLI 720 1175 415 TURBIDITY (Absorbance at 430 millimicrons) 52