ABSTRACT A study was made of the populations of intermediate sized interstitial organisms of Del Monte Beach, Monterey California. In the summer of 1969, this beach was closed because of sewage pollution from the city's offshore outfall. It was expected that populations of organisms constantly exposed to this pollution might be affected. Samples were taken at every 100 meters on a transect runningl000 meters north and 1500 meters south. Counts were substantially lower to the north than to the south. This appears to be correlated to the southerly flow of the sewage as reported by several other investigators. A general enrichment of the environment is considered to account for the increase in population. INTRODUCTION As part of a research project on the sewage pollution of Monterey Bay, a survey was undertaken of the intermediate sized interstitial organisms of Del Monte Beach, fronting the city of Monterey. During the summer of 1969, these beaches were closed due to a high degree of bacterial contamination. It seemed likely that an effect might be observable in populations of organisms constantly exposed to this pollution. The organisms studied were intermediate in size between the protozoa, and such macrofauna of the beach as sand crabs (Emerita sp.). The animals surveyed were nematodes, nemerteans, flatworms, annelids, and harpacticoid copepods. Others which occured regularly but not in surveyable numbers included amphipods and mysids. While work had been done on the general ecology of interstitial organisms, this appears to be the first study done to correlate their ecology to possible pollution. General studies of their ecology include those of Pennak (1951) and Swedmark (1964). METHODS From the location where the pipe from the Monterey sewage treatment plant crosses the beach, running to the offshore outfall, a transect was established with stations every hundred meters for 1000 meters north and for 1500 meters south. The southern transect terminated at Monterey Municipal wharf f2. Sand samples were taken at the same tidal height and stage of the tide once a week for three weeks with a core 8cm deep and approximately 4cm in diameter to produce a sample of 100 cc. The samples were stained and preserved with a mixture of rose bengal and isopropyl alcohol. The organisms were counted by spreading loc sub-samples in Syracuse dishes, adding water, and spinning so that a layer of sand grains was produced over which the stained organisms (which were slightly bouyant) could be easily seen. A count was also taken of the organisms caught when the preservative fluid was filtered off. 24 RESULTS The results of the counts are shown in Table 1. The tabulated values are the means of the three weekly counts. At 100 south and 500 north, additional samples were taken to refine the value of those stations which seemed extremely variable. All counts are innumbers of organisms per l0cc. The third category lumps nemerteans and flatworms together because of the low counts and the dif- ficulty of differentiating preserved individuals of these two similar groups. Graph 1 shows the total number of organisms of all categories per lOcc plotted against distance. The line is the connection of the individual means while the vertical lines are the standard deviations of the samples at each station. Graph 2 is the population of nematodes plotted against distance from the outfall. As can be seen by comparison with the first curve, the nematodes are the largest constituent in the total population, and the variation in the total population is largely a function of the variation of the nematode population. Graph 3 displays the distribution of annelids in the population, primarily a Phyllodocid worm, possibly Eteone dilatae. As can be seen from the graph, the population of this worm increases with distance north of the outfall. The populations of the other organisms are not graphed because they did not appear to show any definable trend over the length of the survey. DISCUSSION A general trend in distribution appears to be evident along the length of this survey. The counts of the nematodes show a major change in the first 100 meters on either side of the outfall pipe. To the south the abundances are consistently high with a variation that could be accounted for by the variation in sampling. To the north, however, the population falls off sharly with increasing distance from the outfall. The population of the annelid worm exhibits a reversal of this trend, increasing with distance north. This would seem to indicate a general ecological variance between the northern and southern half of the survey There are several reasons for believing that the effect is related to sewage pollution. The first is the relative homogeneity of the beach environment along the length of the survey. The second is the abrupt change in the curve at the area of the outfall. The third is the nature of the sewage field around the Monterey outfall. Several ecological factors important to the distribution of interstitial organisms are relatively constant along the length of this beach, or else vary in a manner inconsistent with the observed variation in the population. Station 0 Pipe N 100 200 300 400 500 600 700 800 900 1000 S 100 200 300 400 500 600 700 800 900 1000 1250 1500 Total 116.2 93.5 58.5 46.5 24.1 33.6 60.3 43.2 19.0 39.5 22.3 201.5 77.4 112.6 117.2 178.7 83.1 140.2 89.3 96.0 86.3 95.1 92.5 Nematodes 80.7 48.8 47.3 36.3 13.4 14.7 29.4 15.2 6.8 5.1 6.2 117.6 43.0 87.0 110.6 145.4 62.5 113.9 70.7 74.5 58.0 52.3 78.7 TABLE I Annelids 6.7 5.4 4.7 4.7 4.0 11.7 9.7 9.1 30.0 12.1 3.8 4.6 3.3 2.6 3.6 3.6 5.0 3.6 5.4 5.0 1.6 1.7 Nemerteans Fla.worms 5.2 3.8 2.5 3.7 3.2 3.8 9.5 9.9 1.7 2.3 8.7 2.6 6.3 3.3 6.2 5.2 12.0 8.4 7.7 14.3 11.5 12.0 Copepods 23.6 35.5 1.8 1.8 8.1 9.7 9.0 1.4 1.4 1.7 71.4 27.2 16.0 23.0 11.8 10.7 6.6 8.4 9.0 30.0 248 FIGURE CAPTIONS Figure 1 - Map of Study area showing transect lines. Graph 1 - Total abundance (Number/ l0cc) vs Distance from outfall Graph 2 - Nematode abundance (Number/ l0cc) vs Distance from outfall Graph 3 - Annelid Abundance (Number/ 10cc) vs Distance from outfall Graph 4 - Semi-log Plot Total Abundance (Grouped in 300 meter groups) vs Distance from outfall AaR¬ POINT CASSLO OINT ALONES MONTEREY g. sn MONTEREY +++ Os - MARINA Sad + a sgre / 8 AVE r DEL MONTE 5 e er oserd 1. Sa TAUENER ACMNTR COUNT egos. gean ane un . — POSTGRADU 5 T. THE UE areso. 821 SCHOOL P. sge e aere 1 1 - ee VONTER SENN FRENONT . 8 a 1 Jt CLUS HOUSE, MONTEREY 4 DEL MONTE 2 PENINSULA de g BICKETIS 20 BO LEANV M NAn COLLEGE GOLF 5 18 S 1-------- Kge- rs 2 5 229 2 e 46u SFECHTE DEL MONTE COUE 68 8540 NA 9 — e O 1 S 5 5 245 o - 1 e 10 12 z skakatav- e e I 2 The grain size of the sand is relatively constant along the portion of the beach in question, with most sand grains having a diameter of O.2 — O.5 mm. At 1500 meters south, however, there are large pebbles scattered in the sand. The sand from the ends of the transectare indistinguishable in size, though the sand form the southern end is darker. Temperature (as measured off shore in another survey) varies only 0.2° C along the survey length. Salinity variations offshore also do not appear to be significant It seems reasonable to assume that without any great variation in these factors in the offshore waters, then a similar homogeneity can be expected in this regard along the length of the beach. Other parameters of the interstitial environment such as dissolved oxygen or the chemical composition of the water are very difficult to measure in situ. Zinn (1968) mentions the development and testing of such a measuring device. No such measurements were attempted in this study. The problem of wave action is somewhat more difficult to assess, since there is an appreciable difference in the intensity of wave action along the length of the transect. At the extreme southern end the wave action is slight with the waves being less than O.1 meters in height, while at the farthest north station, waves of O.4 — 0.7 meters are present. As there are no major obstructions or topographic anomalies along the beach, it seems likely that the height and intensity of wave action increases in a continuous and smooth function up the length of the beach. This is in contrast to the population distribution. There is a large field of kelp on a bed of slate to the south of the outfall. The influence of this on the interstitial environment ashore is not known. If the changes of the beach population were independent of sewage pollution, then it would be unlikely that any significant change in the population could be relatable to the presence of the outfall. A semi-log plot of the counts grouped in 300 meter groups in both directions from the pipe shows a definite change in slope in the vicinity of the outfall. This method of presentation indicates more clearly the major trend along the beach by reducing the random variations in individual samples. Studies of the Monterey outfall indicate that some of the sewage comes ashore near the outfall. Trumbauer (1966) shows the highest coli-form counts within 200 meters of the outfall. His report also mentions that the area of highest concentrations is quite variable, with peaks often occuring far to the south. As has been noted before, coli-form organisms have certain limitations when used in sewage tracking. 228. NUMBER 1Occ 150 100 50 25 8 1200 600 PIPE 600 NORTH S. A more informative study was that done by Stevenson (1964) of the water circulation pattern off Del Monte Beach. In 10 surveys, a southward drift was reported six times, a northward drift twice, and on shore drifts, normal to the beach twice. He utilized drogues released seaward from the boil. He concluded that the major factor in water movement in this area is wind. The major wind component in May is from the north 30% ofthe time, northwest 33% and northeast 5%. (Data from U. S. Navy Meteorological Atlas - North Pacific). This wind pattern would give a southerly flow during the month of May. The finding is supported by the dye tracking study and chemical analysis of Blencowe (personal communication) and the diving observations of Baxter and Webster (personal communication). These divers, participating in a benthic survey, report the observation of what appears to be a visible sewage field moving to the south. CONCLUSIONS The result of the sewage pollution produced by the Monterey sewage treatment plant is an enrichment of the populations of interstitial organisms to the south of the outfall where a large amount of the sewage appears to be moving. The sewage could be enriching the beach environment directly by providing readily absorbable nutrients, it could be providing basic nutrients which would promote the growth of bacteria, algae and fungi, or it could be providing a constant supply of bacteria (live or dead) on which the organisms could be feeding. Not enough is known about the viability of fecal bacteria in sea water, the decay of complex organic molecules in seawater, or even about the biology of these particular organisms to conclude which of the enrichment mechanisms is most likely to be in operation. ACKNOWLEDGEMENTS Acknowledgement is gratefully made to Dr. E. H. Wheeler for his advice and encouragement on this project and to Dr. Welton Lee and Dr. Donald Abbott for assisting in the identification of some specimens. This study was supported by grant 617280 of the National Science Foundation. 28. REFERENCES Pennak, R. W. 1951 Comparative ecology of the interstitial fauna of freshwater and marine beaches. Annee biol. ser (3) 27, Ml9-180. Stevenson, C. D. 1964 A study of the currents in southern Monterey Bay M.S. thesis in oceanography. U.S. Naval Postgraduate School Monterey California Swedmark, B. 1964 The interstitial fauna of marine sand. Biol. Rev. 39 1-12 Trumbauer, D. S. 1966 A coli-form bacteria survey of Monterey Bay off Del Monte Beach. M. S. thesis in oceanography. U.S. Naval Postgraduate School Monterey California Zinn, D. J. 1968 A brief consideration of the current terminology and sampling procedures used by investigators of marine interstitial fauna. Trans. Amer. Microsc. Soc. 87 (2) 219-22: 254