Introduction Little is known of the effects of sewage on ma- rine organisms. This study attempts to measure some of the effects on the intertidal hermit crab Pagurus sam- uelis (Stimpson, 1859). Field studies were conducted at the sewer out- fall on the southwest side of Point Pinos, Pacific Grove, California. The city of Pacific Grove (pop. 14,000) dumps into the ocean approximately 14 million gallons of primary-treated sewage per day. The outfall is located about 0.5 feet above Mean Lower Low Water in an area which receives heavy wave action. The sew- age is composed mainly of suburban household wastes, and contains almost no industrial or agricultural wastes. Primary treatment removes grease, oil, and other floatables, and allows some settlement of solids; the treatment plant includes a digester which uses bac- terial action to break down some organic material, obut this was not operating effectively during the study period. Considerable controversy occurred in the fall of 1969 concerning the quality of sewage treament by Pacific Grove and other Monterey Bay communities. The Monterey County Board of Health ordered the closing of many beaches around Monterey Bay in September 1969 when counts showed coliform bacteria levels exceeding those permitted in areas of water contact sports by the State of California. "Cease and Desist" orders were is- sued by the Regional Water Quality Control Board to the city of Pacific Grove and other communities, and later a ban was placed on further sewer connections. Some time after the closing of the beaches, the Paci- fic Grove sewage treatment plant stepped up its chlo- rination in an effort to bring down coliform levels, producing levels of residual chlorine up to at least 45 ppm and an extension of the visibly damaged area around the outfall on Point Pinos by April 1970. A preliminary survey showed hermit crabs to be absent in the area. In subsequent studies, the distri- bution of Pagurus genus was determined, the toxicity of chlorinated and unchlorinated sewage to hermit crabs was studied in both laboraory and field, and the ten- dency of hermit crabs to move away from various con- centrations of sewage in the laboratory was measured. Outside of the distribution study, only P. samuelis was used due to its more ready availability. I. Intertidal distribution of Pagurus in relation to the sewage outfall. Anquantitative study of the distribution of Pa- gurus was made in May,1970 at Point Pinos, and the relative abundance of each of the four intertidal 1/6 species, P. samuelis, P. granosimanus, (Stimpson, 1859), P. hemphillii (Benedict, 1892), and P. hirsutiusculus (Dana, 1851), was determined. Twenty-eight one meter wide transects were ex- amined at low tide at various locations on Point Pinos. Each transect was approximately 10 meters long, begin- ning at the low water level of the ocean at the time -1.0 to 0.0 feet in tidal height). Every hermit crab as it was found was scored as to (1) species, and (2) approximate size, using the width of the base of the shell (always of the turban snail Tegula in adults) as a rough measure of the size of the individual. Adults were identified using Light's manual (Light, Smith, Pitelka, Abbott, and Weesner 1964), and juveniles using Putnam and Markham's key (Putnam and Markham, 1965). Populations in three large tidepools were considered as separate units. For each transect or large pool, the following were determined: (a) the total number of Pa- gurus, (b) the total number of each species, and (c) the total number in each size-class for each species. Three size-classes were defined: large (shell base width greater than or equal to 20 mm; carapace width greater than 7.5 mm); medium (shell base width 10 to 19 mm; carapace width 2.5 to 7.5 mm); and small (shell base width less than 10 mm; carapace width less than 2.5 mm). Areas with suitable Pagurus habitat (pools which do not drain even at lowest tides) were deli- neated. Relative abundance of Pagurus in each area was scored as follows (see Fig. 1): abundant (over 100 individuals per transect or 10 m2); fairly common (20- 99 per transect or 10m2); scarce (less than 20 per transect or 10-m2); absent (no individuals eseened) Areas where no transects were taken were examined qua- litatively. Location of transects and totals for each species and size class for the transects are given in Appendix A; areas containing suitable habitat and the relative abundance of the genus in each area is given in Fig. 1. No alterations in the pattern of distribution were ob- served where the genus was present: in areas below +1.0 feet in tidal height, large P. granosimanus and small P. hirsutiusculus were by far the most common, and above t1.0 feet, P. samuelis was the only species found, with all three size classes being common. The general distribution of the genus in relation to tidal height and the relative abundance of the size classes conformed to that found by Belknap and Markham (1965). In only two areas was Pagurus abundant, and in most areas the genus was classified as scarce. Pagurus was absent within about a 100 foot radius of the out fall, although the abundance of pools within this area would normally make it highly suitable for Pagurus. Al. gal coveri is greatly reduced or lacking within this 100 foot radius. Pagurus was also absent in two areas of Cove A. Areas where hermit crabs were abundant or fairly common were as close to the outfall as 120 feet, just outside the area where they were absent. The heterogeneity of the intertidal makes it difficult to say much more about the distribution of Pagurus at Point Pinos than that they were absent within a 100 foot radius from the outfall, and that Pa- gurus isgenerally not very common at Point Pinos. In no area was the density of Pagurus as high as that found at Mussel Point approximately two miles southeast of Point Pinos. II. Field studies on the toxicity of sewage to Pa- gurus samuelis The toxicity of sewage under field conditions over a one month period was determined by placing wire cages containing P. samuelis at different distances from the outfall (Fig. 1). A control was placed at the Hopkins Marine Station on Mussel Point, a locality con- sidered to be unpolluted. Cages were constructed of stiff, 4 in, mesh, galvan- ized wire screen molded into flattened cylinders about 60 cm long, 15 cm wide, and 10 cm high. Twenty-five P. samuelis collected at Mussel Point were placed in each cage: 6 large, 14 medium, and 5 small. The small ones were placed inside a separate envelope of plastic screening 12 cm by 5 cm. Rocks were added to the cage for weight. A dead gooseneck barnacle (Pollicipes po- lymerus) was added to each cage for food at weekly in- tervals, although Richardson (1965) found these crabs can live for 30 days without food with only 7.6% mor- tality. The density if the animals in the cages was roughly half that calculated by Richardson (1965) to give less than 20% mortality in 5 days. On 1 May 1970 11 cages were placed at Pt. Pinos in pools where they were always submerged. All were below a tidal height of 0.0 feet except cage 4. The cages were wedged un- der or between rocks in the pools and were not moved by wave action. Cages were examined every other day when tides permitted; the longest period between successive ob- servations was 7 days, but this came after the period when most mortality occurred. The number of dead indi- viduals in each cage was recorded at each observation. An individual was considered dead if (a) an empty shell but no body was found, or (b) the animal showed no resistance to being removed from its shell and showed no response to prodding after 10 minutes in fresh sea water. The percentage of surviving animals at each ob- servation is given in Fig. 3. and the time at which 50% mortality (Lethal Time 50) occurred in relation to distance from the outfall is given in Fig. 2. All animals died within 9 days in cages within a 100 foot radius of the outfall, death occurring most rapidly in cages closest to the outfall. Mortality in cage 6, 110 feet from the outfall, reached 72% in the 28 day ob- servation period. The remaining cages, including the control, showed 8 - 16 % mortality in 28 days, which is approximately the same as in controls maintained in the laboratory (see below). Cage 11 was destroyed by children. A very short LT 50 was found in cage 4, despite its relatively great distance from the outfall. It was 12 situated in a high tidepool (13 feet in tidal height) rather than below 0.0; polluted water is thus trapped here for longer periods of time than at lower levels. The field cage study shows that any P. samuelis within a 100 foot radius of the outfall were killed but beyond 110 feet there were no lethal effects in four weeks. This correlates with the distribution study, which showed no P. samuelis within the 100 foot radius. A high mortality did occur in cage 6, where large numbers of living P. granosimanus and P. hirsu- tiusculus were found; however, the living crabs were concentrated at the end of the pool farthest from the outfall, and the cage was at the nearest end, where few living Pagurus were found. Field observations show that murky water from the outfall begins to seep into the pool as the tide rises to about 1.0 ft. in tidal height; thereafter it is flushed out with clean water from the opposite direction as the tide reaches about 13.0 ft., and heavily polluted water is not concentrated in the far half of the pool. In a similar field cage study conducted by the author in August 1969, no significant mortality occurred in any cages within the 100 foot radius in 12 days. The closest cage to the outfall was 15 feet away. This was before heavy chlorination, thus implicating chlorine as the factor producing high mortality in May 1970. III. Laboratory studies on the toxicity of sewage to Pagurus samuelis. Experiments to determine the short-term tox- icity of sewage to P. samuelis in the laboratory were carried out as follows: In each test 25 P. samuelis (5 large, 10 medium, and 10 small) were placed in shallow plastic dishes each with a bottom area of about 600 cm2. This is the approximate density calculated by Richardson (1965) to give less than 20% mortality in 5 days (25 aniamls per 525 cm), although only 8% mortality in 29 days occurred in controls in this experiment. Occasionally larger dishes with a bottom area of 1000 cm were used, and the number of individuals in these dishes was raised to 40 to acheive the same density. The animals never utilized all the area available to them, clustering in the corners most of the time; thus crowding was not thought to be a critical factor. Each group of animals was given a 24 hour acclimation period in fresh sea water before antest, since Richardson (1965) states that most mortality-producing interactions occur in the first four hours. In 25 experiments, however, only 2 out of 705 hermit crabs were found dead after the ac- climation period. 16 10 Fifteen-hundred ml of solution were added to the smaller dishes and 2000 ml to the larger ones, gi- ving a depth of about 1.5 inches, in which the animals were continuously submerged. No aeration other than diffusion was used since the surface to volume ratio was large. Dishes were placed in a circulating sea water bath to maintain a constant temperature of about 14°. This is slightly higher than the average sea water temperature during May in Monterey Bay (12°0), but is well within the temperature ranges if the high tidepools in which P. samuelis abounds, and is well below thermal death point for P. samuelis (31.500; Forward, 1965). The following solutions were tested: (a) chlorinated Pacific Grove sewage at con- centrations of 100%,50%,20%,10%, and 1%. Chlorine levels in straight sewage ranged from 15 to 45 ppm as determined by idiometric titration as given in Standard Methods for the Examination of Waste and Wastewater,1966. Residual chlorine levels were found to drop by about 25% after 6 hours after arrival from the sewage plant. No chlorine was ever detected in the 1% concentration, but the assay was not sensitive in the levelseexpected at that concentration. (b) Pacific Grove sewage at concentrations of 100%, 50%, 20%, 10%, and 1%, but dechlorinated with 11 sodium thiosulfate, Na2S203, to determine if chlorine was a toxic factor; 0.36 g of thiosulfate were added for each liter of sewageiin the solution. Thisiwas enough to neutralize approximately 100 ppm chlorine, more than twice the amount ever detected in the sewage. (c) a control for (b) alone using sea water and sodium thiosulfate at 0.36 g/liter, the highest concen- tration in any solution, to determine if thiosulfate itself could be toxic. (d) 100% Pacific Grove sewage dechlorinated with O.36 g/liter thiosulfate and raised to the salin- ity of sea water by adding 33.5 grams of "Instant Ocean" salts per liter of sewage. (e) unchlorinated sewage from the treatment plant of the adjacent city of Monterey in concentrations of 100%,50%,20%,10%, and 1% to compare the relative toxicities of sewage from the two areas. (f) distilled water at concentrations of 100%, 50%,20%,10%, and 1% for an osmotic control. (g) tap water at 100% concentration. Tap water is closer to the salinity ofsewage than distilled water. (h) 100% sea water control. Two tests instead of one were meade for each 100g solution showing toxicity (50 animals). The larger dishes (40 animals) were used for all 50% concentrations. All dilutions were made using sea water from 2 12 the sea water system at Hopkins Marine Station. One or two gooseneck barnacles (Pollicipes polymerus) were fed to each group once a week. Uneaten portions were removed after 18 hours to prevent fouling of the water. As mentioned above, food was not considered a critical factor in mortality (Richardson, 1965). Fresh sewage was obtained every morning from the Pacific Grove, and Monterey sewage plants, and test solutions were changed within 2 hours thereafter. Solutions not con- taining sewage were also changed daily. Each test solution was examined at intervals determined by the relative toxicities of the solutions, i.e. hourly if highly toxic or twice daily if no rapid mortality observed. Criteria of death were as follows. Individuals showing no response to prodding were re- moved and placed in fresh sea water; if after 15 mi- nutes there was no response to prodding and if the mouth parts were not moving, the individual was con- sidered dead. Often individuals showing no response in the test solution showed limited movement of appendages within 5 minutes exposure to fresh sea water; but twice about 10 animals scored as dead were left for twelve hours in fresh sea water without recovering. The percentages of surviving animals versus time for 100% and 50% solutions appears in Figs 5 & 6. The LT 50's for chlorinated Pacific Grove sewage are 6 given in Fig. 4 (compare with mortality in field cages, Fig. 3). The only solutions below 50% concentration showing mortality significantly greater than the con- trol (8% in 29 days) were 20% and 10% Pacific Grove chlorinated sewage. The duplicate runs in 100% concen- trations showed virtually identical rates of death. All three size classes showed very similar mortality rates in toxic solutions. Appendages of individuals killed in high con- centrations (100%,50%,20%) of chlorinated sewage were constricted, whereas in 10% chlorinated sewage and all other types of toxic solutions, the appendages at death were relaxed. When high mortality occurred in solutions with LT 50's greater than 100 hours, the bristles on the appendages were whitened, extended, and prominent, with much adhering particulate material. Death in so- lutions with mortality in the range of the control ap parently resulted from predation, although the actual act was never observed; a half-eaten body, a few appen- dages, or an empty shell provided evidence of the cause of death here. Never was a crab found just sit- ting dead in its shell in non-toxic solutions, and very seldom were half-eaten crabs found in toxic so- lutions. No interest in food was shown in solutions with LT 50's under 30 hours. From the laboratory studies it can be seen that chlorine is the most toxic component of sewage, since 27 chlorinated sewage was the only solution tested which caused significant mortality in concentrations below 50%. When chlorine was neutralized with thiosulfate, no significant mortality occurred in concentrations below 50%. Fresh water also contributes to the toxicity of sewage, for when dechlorinated Pacific Grove sewage was raised to the salinity of sea water, its LT 50 in creased from 13 hours to 27. Also, significant mortal- ity occurred in 100% and 50% distilled water and 100% tap water (see Figs. 5 & 6). Unfortunately tap water concentrations below 100% were not tested, but the LT 50 for pure tap water (18 hrs.) was significantly higher than that for pure distilled water (10 hrs.), which has a lower salt content. While both chlorine and fresh water are toxic components of sewage, there are still other toxic factors, since when the chlorine was removed and the salinity raised, mortality still occurred, although at a lower rate. Also, distilled water proved much less toxic at 50% concentration than the same concen- tration of dechlorinated sewage. Since in laboratory experiments sewage solutions were changed once a day, the situation here differs somewhat from that in the field: near the outfall at high tides fresh sewage is added continuously to the 12 area, and only at lower tides would an animal be ex- posed for prolonged periods to the same sewage solution. No studies were made of the changes occurring in sew- age with time other than to note the decrease in chlo- rine content. In any event, since the composition of effluent at the outfall probably varies widely depen- ding on the time of day, it would be difficult to si- mulate in the laboratory the exact conditions which animals encounter in the field. Another factor to be considered is that some areas in the field may receive concentrated doses of sewage for only short periods of time, and the effect of this was not tested in the laboratory. From the laboratory studies it can be seen that the rapid mortality rate noted in field cages placed around the outfall is probably due to chlorine, since the concentration of sewage without chlorine necessary to produce the LT 50 found at 60 feet would have to be greater than 50% (see Figs. 3 & 4). The LT 50 at 60 feet from the outfall (96 hrs.) more closely corre- sponds with that for 10% chlorinated Pacific Grove sewage (91 hours) than that of 50% dechlorinated Pac- ific Grove sewage (212 hrs.). Most components of the sewage were found to be diluted to 20% or less at 60 feet (Hopkins Marine Station chemical study group, pers. comm.), and thus mortalities at this distance are more likely caused by a 10% concentration than a 50% concentration. As much as 6.9 ppm residual chlorine was detected 60 feet from the outfall on some days, al- though none was found on others (Elaine Anselmo, pers. comm.); this is about 20% of the average chlorine con- tent of the effluent. 17 IV. Movement of P. samuelis away from sewage It was hypothesized that the absence of P. sa- muelis in the outfall area might be accounted for by movement of the animals away from the outfall, and so an experiment was devised to measure movement out of various concentrations of sewage. Plastic boxes l0cm by 10 cm and 6 cm deep were clamped togehter in pairs joined by a strip of plastic screening extending from the floor of one box to the floor of the other. The hermit crabs could thus easily climb out of one dish and into the other, using the screen as ladder and bridge. The boxes were filled with test solutions to within 5 mm of the top (about 500 ml). Each pair of boxes contained sea water in one and test solution in the other. Controls were run with sea water in both boxes. The sewage was brought to sea water temperature before use (from about 18°c to 14°c). During the experiments the solutions warmed gradually to about 16°0. No aeration was used. The following solutions were tested, each at 100%,50%,20%,10%,and 1% concentrations: (1) unchlori- nated Pacific Grove sewage, (2) unchlorinated Pacific Grove sewage raised to sea water salinity by adding 33.5 g "Instant Ocean" salts per liter sewage, and (3) tap water for an osmotic control. 18 Five large or medium P. samuelis collected in unpolluted pools at Hopkins Marine Station were placed in every box at the start of each experiment, and their positions recorded after one hour. Ten trials were run with each dilution of each solution, to give 100 in- dividuals for each test. New crabs were used in each- trial. Almost as soon as crabs were placed in the con- tainers, rapid movement occurred between the two solu- tions. Only 5% of the animals tested never left their original boxes. The plastic screen bridge was easily climbed, but a substantial percentage of animals pre- ferred to remain on the bridge rather than stay in either solution. Most movement ceased in 45 minutes. The percent of individuals in each box and on the bridge after one hour is given in Table 1. What was considered important was the ratio of the number of individuals in the sea water box to those in the test solution box; this is given for each test in Table 1 and plotted against concentration of solution in Fig. 7. The experiment did not work with higher con- centrations of chlorinated Pacific Grove sewage, since most animals were incapable of climbing the bridge af- ter 5 minutes in 100%,50%, and 20% concentrations. Only one of 30 animals tested remained in 10% Pacific Grove chlorinated sewage after one hour, but no pre- ference was shown between 1% Pacific Grove chlorinated sewage and sea water. In the movement experiments, a solution was con- sidered to produce movement away from it if the ratio of individuals in the sea water box to those in the test solution after one hour was greater than 2:1. The three controls averaged 1.2 : 1, fairly close to the l:l ratio expected by chance alone. On this criterion, movement shown seems to be related mainly to reduc- tion in salinity; sewage and tap water dilutions showed approximately the same ratios, but sewage so- lutions adjusted to sea water salinity by addition of sea salts produced no movement except in 100% concen- tration. Movement away from 100%, 50%, and 20% so- lutions of both sewage and tap water was recorded, but not at 10% and 1% concentrations. If movement away from an area is responsible for the absence of P. sa- muelis there, the experiments suggest the animals move cif the salinity is at or below 2700/00, but nothif higher than about 30 0/00. Since salinities measured were lowerröthan 30 0/00 in the field only within a 25 foot radius of the outfall, it seems likely that the absence of P. samuelis within a 100 foot radius is not due solely to movement away from the pipe. Summary 1. Pagurus is absent within a 100 foot radius of the Pacific Grove sewage outfall. No abnormalities in the patterns of distribution of Pagurus were found outside the 100 foot radius. 2. Field cages containing P. samuelis placed within a 100 foot radius of the outfall showed 100% mor tality in 28 days. Beyond this distance, mortalities did not exceed those in unpolluted areas. 3. Chlorinated sewage (residual chlorine 15 - 45 ppm) was the only one of several solutions tested in the laboratory (including dechlorinated sewage) showing significant mortality in 20% and 10% concentrations. Chlorine was probably the cause of deaths recorded in field cages within 100 feet of the outfall; lab- oratory studies showed sewage without chlorine could not account for this mortality except in concentra- above 50%. 4. Fresh water is a component of sewage toxic to P. sa- muelis at salinities of 0 - 17 0/00, but not toxic at 27 0/00 and above in115 days. 5. Other toxic components are present in sewage since mortality exceeding that of controls was recorded in sewage which was dechlorinated and osmotically bal- anced. 1 6. P. samuelis tends to move away from solutions with salinities at 27 0/00 or below, but this alone pro- bably cannot explain the absence of this species within the 100 foot radius of the outfall. P. samuelis is not a sensitive indicator for un- chlorinated sewage, but is very sensitive to chlor- inated sewage. If sewage is not chlorinated, P. sa- muelis could survive for prolonged periods in 20% concentration of sewage 22 Acknowledgments These investigations were supported in part by the National Science Foundation Undergraduate Re- search Program, Grant no. GY-7288. I wish to thank Dr. Donald P. Abbott for his help and suggetsions during the experiments and his invaluable criticism of the manuscript. I would like to thank James Schreiber for making his chlorine testing apparatus available to me. Elaine Anselmo provided chlorine data from the outfall area, and Victor Anderlini, Elaine Anselmo, Dave Clear- man, John Hainsworth, Marshall Holstrom, and Tom Rot- kis cooperated with me in obtaining chemical data at Point Pinos. Marshall Holstrom provided the Pollicipes used for food. 23 References 1966. Standard Methods for the Analysis of Waste and Waste Water. Am. Public Health Assoc. New York. pp 376 - 378. Pers, comm: unpublished data from Point Pinos chemical study group. May 1970. Elaine. Pers, comm: chlorine levels at Anselmo, Point Pinos. May 1970. Robert and John Markham. The intertidal Belknap, and subtidal distribution of four species of Pagurus (Fabricius) at Mussel Point, California. Unpublished student report, 1965. Hopkins Marine Station, Stanford University. pp. 20 - 43. Richard. 1965. Effects of temperature and Forward dessication on Pagurus samuelis and Pagurus granosimanus at Pacific Grove, California. Unpublished student reports, Hopkins Marine Station, Stanford University. pp.126 - 150. Light, S.F., Ralph I. Smith, Frank A. Pitelka, Donald P. Abbott, and Francis M. Weesner. Intertidal Invertebrates of the Central California Coast. Berkeley, 1964. pp. 183, 244, 245. John D. and John C. Markham. 1965. Charac. Putnam, teristics of larval and postlarval stages for Pagurus in Monterey Bay, California. Unpublished student reports, Hopkins Marine Station, Stanford University. pp 198 - 217. Richardson, Norman. 1965, Some factors of mortality in laboratory populations of Pagurus samuelis. Unpublished student reports, Hopkins Marine Station, Stanford University. pp 218 - 230. Figure 1 Distribution of Pagurus at Point Pinos and location of field cages. Numbers indi- cate position of field cages. Areas deli- neated contain suitable Pagurus habitat. 2- outfall COVE A 12 at-HOPKINS MARINE o 0 a E Figure 2 Relation of LT 50 to increasing dis- tance from outfall for field cages. Numbers refer to specific field cages given in Fig. I. Dashed line indicates distance beyond which no significant mortality ogcurred. Cage 4 was in a pool 3 feet higher than the other cages. 9. SAOOH 0g O LL L LE Z Figure 3 Per cent survivors as a function of time for animals in field cages at Point Pinos. Numbers refer to specific field cages as given in Fig. 1. Cage 4 was in a pool 3 feet higher in tidal height than the other cages. 9. 0 O SAOAIAAOSINd 8 V. Figure 1 Per cent survivors as a function of time for Pagurus samuelis exposed to chlorinated Pacific Grove sewage. Letters indicate concentration: A - 100% B 50% C - 208 10% 18 F- sea water control 1 SAOAIAAOS INSD Aad 1 1 Figure Per cent survivors as a function of time for Pagurus samuelis exposed to 100% solutions. Letters indicate type of solution: chlorinated Pacific Grove sewage b- distilled water unchlorinated Monterey sewage dechlorinated Pacific Grove sewage tap water dechlorinated and osmotically ba- lanced Pacific Grove sewage. g - sea water 9. 2 SAOAIAAOS INA a 18 Figure Per cent survivors as a function of time for Pagurus samuelis exposed to 50% concentrations of test solutions. Letters indicate type of solution: a - chlorinated Pacific Grove sewage distilled water unchlorinated Monterey sewage dechlorinated Pacific Grove sewage pure sea water control * 00 1 SAOAIAAOSIND ad Table 1 Movement of P. samuelis away from sew- age: Per cent individuals in each box after one hour. All boxes began with 5 in sea water, 0 on bridge, and 5 in test solution, and 10 runs were done for each concentration. A - unchlorinated Pacific Grove sewage B - unchlorinated and osmotically ba- lanced Pacific Grove sewage. C - tap water. A B C Concentration ofsewage 1008 507 204 10 19 sea water control Concentration ofsewage 100% 505 204 0% sealwater control Concentration water of tar 100% 504 258 163 12 seanwater control sea water 29 sea water 12 sea water 67 58 55 38 bridge bridge 28 32 bridge 28 29 sewage 44 sewage 127 120 39 27 tap water 16 142 116 Tatio S.Wse 112 ratio s.w./sew. 918 2 ratio s.W./tap 13.4 3.4 Toble 15 Figure 7 Movement of P. samuelis away from sew- age : per cent concentration of test solution ys. ratio of individuals in sea water box to individuals in test box. A - unchlorinated Pacific Grove sewage B - unchlorinated and osmotically ba- lanced Pacific Grove sewage. C - tap water 95 u uoljojos 1891 10 ouu ue5 le 8 — .2 9 Appendix A Location of transects for Pagurus di- stribution study at Point Pinos. S OUTFALL 50 100 feet — V — POOL Pool A 01716 1514 o W 22 24 26 28. 25 20 21 a 06 U Aepend /88 Appendix E Numbers of Pagurus found on each tran- sect in distribution study at Point Pinos. Numbers refer to transect number in Appendix A, and letters refer to pools in Appendix A. 4 L L Z T « - ap; 2 10 11 12 14 15 17 18 19 20 21 22 23 24 25 26 27 28 B 2 0 — 12 T a 0 13 7 40 27 a 29 0 — 28 20 21 39 48 57 ao 11 15 12 11 12 25 17 4 OIAIO TaodLaa 0 22 48 682 188