Vertical Distribution of L. scutulata Merchant, Noroian Introduction The small checkered periwinkle, Littorina scutulata (Gould, 1849), is prevalent in the upper intertidal zone of the California coast. It is found to inhabit tidepools as well as cracks and rock faces in this zone. Previous work on movement and migration of L. scutulata is not abundant. Hewatt (1937), demonstrated negative photo- taxis in this species. North (1954), observed that differ- ences in the degree of exposure to wave action appear to influence distribution of the animal within intertidal zones. Glynn (1965), further observed that vertical movements in the Balanus-Endocladia zone are made with the rising and falling tides, upward movement coinciding with the advancing tide. Preliminary field observations of L. scutulata in tide- pools suggested the existence of rhythmicity in migration into and out of these pools. The purpose of this study was to obtain a clearer picture of vertical distribution in tidepools over time and to look for possible factors influ¬ encing this aspect of behavior. Study Areas Observations in the field were carried out at Mussel Point, Pacific Grove, California. Two areas of differing Vertical Distribution of L. scutulata Merchant, Noroian geography providing different degrees of exposure to incoming tides were studied. In each area two pools were observed. See Figures 1 and 2. Area I is located on the north side of the Point on a flat, horizontal rock surface. There are numerous large rocks in the lower intertidal region which act as a break and greatly lessen the impact of waves at high tides. Waves tend to roll over the pools, not crash over them. Area II is on the west shore of the Point and is exposed to much stronger waves then Area I. The pools are on the seaward edge of a horizontal plateau. They receive much spray and splash at high tides. Pools 1 and 2, located in Area I,are connected at high tides by a narrow channel which fills with water. Pools 3 and 4, in Area II, are adjacent and separated by a large crack. Pool 4 is connected with another pool at one end. All pools are granite, covered with a thin film of algae. Macroscopic algae is found only in Pool 1 which has two small bunches of Rhodoglossum affine. There are no rocks or sediment in the pools and their water is not turbid. The pools vary greatly in depth. Pool 2 is the shallow¬ est with a maximum depth of 3 cm. Pool 1 is the deepest with a maximum depth of 8 cm. Pools 3 and 4 have intermediate depths of 5 cm and 7 cm respectively. Two studies were carried out in Areas I and II. The first consisted of two 60 hour observations of gross population movement. The second was a 53 hour investigation Vertical Distribution of L. scutulata Merchant, Noroian of the movement of individual snails. Materials and Methods A photographic sampling technique was used. A one-half meter square quadrat, divided into one-hundred 5 cm x 5 cm squares, was placed over a pool in a stadardized position. A photograph was then taken on Ecktachrome64 color positive film using a Nikon F2AS camera and either a 105 mm or a 55 mm Nikkor macro lens. To minimize parallax the photograph was taken from the same position each time. At night a Vivitar 292 electronic strobe with a duration of approximately 0.OOl seconds was used as a light source. Slides were projected on a screen with a Kodak slide projector using a Kodak Ektanar zoom lens. Counts were made from the projected image. The photographic sampling technique was chosen for two reasons. First, it minimized disturbance of the snails and the area, especially in terms of exposure to light during the night. Second, it recorded distribution at a point in time rather than over the period of time necessary for observations by eye. While replicate counts of the slides significantly differed between the two investigators, their combined counts give an error of less than 6% at the 95% confidence level. A possible problem with the method is the inability to resolve small snails, biasing the study towards the behavior of larger individuals. White fingernail polish was used to mark snails with Vertical Distribution of L. scutulata Merchant, Noroian numbers or dots. A toothpick was used to apply the nail polish. Only the largest snails, greater than or equal to O.5 cm, were used. Results The first 60 hour study was done April 28-30, 1978; the second was done May 16-18. The two studies were per- formed 18 days apart in efforts to separate the tidal and diel components of the changes in L. scutulata distribution. The weather and surf conditions were also different. The first study period was balmy and overcast until late afternoon. The surf was mild. The second study period was clear and warm but the surf was high, calming down slightly the second day. The fluctuations in population distribution were monitored between two categories, In Pool and Out of Pool. In Pool indicates snails which are completely submerged. The Out of Pool category includes snails completely out of water and at the air-water interface. These categories were selected because fluctuations between them appear to be the sharpest change in behavior which the snails exhibit. Large fluctuations in both distribution and total population size of L. scutulata were observed in all of the pools during both observation periods. The results for the individual pools are presented in Figures 3 and 4. The percent of the population Out of Pool is shown by the solid line. The majority of the individuals in the Out of Pool classification were found at the interface. The dashed line Vertical Distribution of L. scutulata Merchant, Noroian represents the population size. Notes on the amount of splash and wash throughout the observation period were only collected during observation period two. Three generalizations can be made about the fluctuation in percent Out of Pool. These hold for all pools and both observation periods. First, there is consistently a large percentage of snails Out of Pool during descending tides at night. Second, there is consistently a small percentage Out of Pool during daytime low tides. Third, more snails tend to be Out of Pool during the night than during the day. Large percentages are also frequently found, but not consistently, Out of Pool at ascending and high tides. They are seldom found at daytime descending tides. Each pool has its own pattern of fluctuations. The snails in Pool 1 come out of the pool with the receding tide at night and stay out. Most have returned to the pool by the first daytime low or receding tide. Pool 2 snails show similar behavior except that a small percentage migrate into the pool at the nighttime low only to come out again as the tide comes in. During the second observation period a very interesting aberration occurs when the percent Out of Pool peaks sharply at the afternoon ascending tide. Pool 3 snails exhibit behavior very similar to that of snails in Pool 2 during the second observation period; however, it is less regular. Pool 4 shows the most aberrant behavior. Distribution fluctuates with little regularity during the first observation period and with Vertical Distribution of L. scutulata Merchant, Noroian regular peaks at descending tides during the second observa¬ tion period. When the data for all the pools are combined, what appears to be regular movement out of the pool at night and into the pool during the day is apparent. See Figure 5. In order to follow the movements of individuals, all snails were removed from the vicinities of Pools 2 and 4. The total populations of these pools were approximately 130 and 150 respectively. Ten snails selected from the Pool 2 population were numbered 1 through 10. Fifteen were numbered from the population in Pool 4. In addition, twenty snails from each pool were marked with a white spot on the tip of the shell. Labeled snails were returned to their respective pools. Observations were made over a 53 hour period at major points in the tidal cycle and the midpoints between them starting four hours after the snails were returned to their pools. Four categories were used to score the position of individuals at Pool 2. In Pool signifies thoseindividuals totally submerged, as before. Interface signifies those found at or within 1-2 cm above the air-water interface. Snails at the edge of the pool with their shells protruding out of the water were also included in this category. Out signifies those individuals not within the normal confines of the pool and more than 2 cm from the edge of the water. Crack is a separate category, not exclusive of the others and represent¬ ing snails found residing in cracks inside or outside of the pool. The same categories were also used for Pool 4 vertical Distribution of L. scutulata Merchant, Noroian with the exception that In Pool was divided into two sub¬ categories: Bottom, for those found at the extreme depths of the pool, 4.5-6.0 cm below the surface, and Side, for those occurring between the interface and 4.5 cm under the water. The observation period lasted from June 1-3, 1978. The weather was unusually calm. It was overcast both days except for late afternoon of the first day and most of the afternoon of the second day. There was little wind and the surf was calm. Although it was a spring tide period, the pools received little spray or wash except at high high tide. All snails remained in their respective areas except for two of the numbered ones in Pool 2 which were lost. Signif¬ icant migration of marked individuals occurred during the observation period. 34% moved from their pool to nearby pools or cracks. After 50 hours, 10 individuals from Pool 2 had migrated to Pool 1. Similarly, 6 individuals moved from Pool 4 into the pool contiguous with it. Some back and forth movement was also observed in both areas. An influx of unlabeled animals was also observed. A maximum of 11 and 40 unmarked snails were found in Pools 2 and 4, respectively. In Figures 6-9, the number of L. scutulata found at different areas at each pool are compared with the tidal and diurnal cycles. Figure 6 shows the number of snails at the interface plus those out of the pool. In general, peak numbers occur during low tide periods. The highest peaks occur at low Vertical Distribution of L. scutulata Merchant, Noroian tides in the afternoon at both pools. At Pool 2 this represents from 71-82% of the population. At Pool 4 it is from 23-50%. Figure 7 is a comparison of the number of individuals at the interface of the pools. As in Figure 6, peaks occur at low tides in Pool 2. Similarly, the peaks at low tides occurring during the day are much higher than for those occur¬ ring during the night. This represent from 43-61% of the total population. As might be expected, both numbered and dotted subpopulations exhibit the same behavior. At Pool 4 there is no clear pattern of movement to the interface and the number found there is very low in comparison with the total population. The number of snails observed in cracks is shown in Figure 8. At Pool 2 movement to this category does not show any regularity, though there are slight increases in the crack population at high and low points in the tidal cycle. At Pool 4 there appears to be an initial correlation between the crack population and the tidal cycle with a large peak, 65% of the total, occurring at the first high high tide and another, 43% of the total, occurring just before the second high high tide. Figure 9 compares numbered individuals found on the bot¬ tom and side of Pool 4. Most of the numbered snails were submerged during the observation period. In general, the bottom population increases at ascending and high tides. Although the sample size is small and fluctuations represent only a few individuals, a constant population was under Vertical Distribution of L. scutulata Merchant, Noroian observation. In general, individual movement appears to be the same as that of the larger population. This can be seen most clearly in Pool 2. See Figures 10 and 11, where the movement of individuals is shown and compare with other figures. The same picture for Pool 4 is not as clear. There is much more variability in individual movement. Discussion Changes in the vertical distribution of L. scutulata appear to be related to the tidal cycle. These changes are modified by the day-night cycle. This is as expected since movement on their substratum is positively correlated with dampness (Kops, 1964), a function of both tide and the diel cycle. Ascending, high and descending high tides are all associated with wetness as a result of splash or spray from the waves. These three periods differ in amount of wetness and intensity of wave shocks. High tide is by far the most severe and wettest of these periods, followed by ascending, then descending tide. Wave shock can present a substantial danger by dislodging L. scutulata from the rocks (Peterson, 1964). Therefore, it may be important when considering the behavior of L. scutulata and its distribution. During the day the rocks are dried more quickly when not exposed to constant splash. Decreases in the Out of Pool population during high tide 10 Vertical Distribution of L. scutulata Merchant, Noroian in the first two observation periods suggest either movement into the pool for protection or removal by wave action. During the first and calmer period of observations there was no decrease at the peak of the high tide. The amount of movement Out of Pool during ascending and descending tides appears to be influenced by two factors. The first is severity of wave shock and the second is phase of the diel cycle. The high surf during the second observa¬ tion period is correlated with a suppression of the ascending tide peaks but not the descending tide peaks at Pool 4 which is the most exposed. Perhaps both the incoming and high tides were too rough for movement out of the pool. The descending tide became the only available wet period which allowed "safe" movement. Daytime is correlated with complete suppression of the peak at descending tide and only partial suppression of the peak at ascending tide. Coming out of the pool as the tide recedes during the day could be particularly dangerous because the sun can quickly dry the substratum and leave the snail faced with problems of desiccation. There are two other possible explanations, not associated with wetness, for a peak in the Out of Pool number during an afternoon ascending tide. This peak occurs regularly only at the shallowest pool, Pool 2, and only during the warmer observation period. These factors, combined with the lack of wash during the day may have resulted in temperatures exceeding the tolerance range of L. scutulata. This may have caused movement to the 11 Vertical Distribution of L. scutulata Merchant, Noroian interface where evaporation results in a decrease in temper¬ ature. The other possible explanation is that a sharp peak occurs closer to high tide and was not detected due to the length of interval between observations. This possibility seems unlikely because it is the presence of the peak rather than its absence which is the aberration. However, a study on short-term fluctuations where data were collected every 15 minutes for one and one-half hours indicated that signifi¬ cant changes in distribution do occur in times as short as 45 minutes. The effects of tide on L. scutulata appear quite variable. These effects depend on roughness of surf, degree of exposure of a pool to the surf, and even the particular geometry of the pool. Day and night, on the other hand, affect all the pools in the same way. There is less danger of desiccation at night and more during the day. When the gross population distribution data for all four pools are combined as in Figure 5, the interpool variability results in a loss of obvious tidal effects. At the same time, the effects of day and night become even more obvious. It was hypothesized above that L. scutulata may move to the interface during the late afternoon when heat from the sun would increase the temperature of the pool to an un¬ comfortable level. This could occur if the pool did not receive any water during a high tide occurring during the day and if the pool were shallow enough to allow a significant 12 Vertical Distribution of L. scutulata Merchant, Noroian rise in temperature. It has been observed that there is a significant temperature differential between the pool, inter- face, and surrounding rock. The temperature of Pool 2 at the interface was found to be 6° C less than the pool or sur¬ rounding rock in the late afternoon on a warm day when the sun was out for only a few hours. This idea, that the snails use the interface as an air conditioner at times of heat stress, finds support in observations of individual movement. There is a significant increase in the number of snails at the interface during the late afternoon in Pool 2 which has a maximum depth of 3 cm. This pattern is not seen in Pool 4 which has a maximum depth of 7 cm and a lower pool temperature than Pool 2. These observations are not conclusive. More information on response to heat in L. scutulata and temp- erature fluctuations of pools is needed. It has been noted in Pool 4 that some individuals tend to go to the bottom of the pool at times correlated with high tide. At Pool 2 this is not so apparent. Pool 4 is exposed to much greater wave shock at high tide than Pool 2 and it is possible that snails either seek the depths of the pool for protection or that they become dislodged during high tide and are sent to the bottom of the pool. In order to better understand this phenomenon more must be learned about the ability of L. scutulata to hold on during wave shock. Other questions in this regard are: How fast do dislodged snails recover and attach themselves? Does attachment to the bottom provide greater protection than attachment at the 13 Vertical Distribution of L. scutulata Merchant, Noroian interface or elsewhere? In Pool 4, the majority of the marked population was in cracks during the first high high tide. Movement into and out of the cracks then showed a rhythmicity associated with the tide; however, during subsequent high high tides a decrease in the peak number and a shift towards ascending tides occurred. It seems reasonable to hypothesize that the snails seek cracks during high tides as cracks are protected areas which afford shelter from wave shock. Hence, the rhythmic movement described above may be a general response to disturbance, anticipating rough conditions. Removing the snails, marking them, and replacing them may have provided this disturbance. Since it was not very rough during the period of observation, the rhythmic response eventually damped out and "normal" behavior resumed. The general pattern of movement observed in the study with marked individuals, i.e. an increase in the Out plus Interface population at low tides and a corresponding decrease at high tides, is different from that observed in the first study. There are several possible explanations for this difference. The results obtained in the last study may be biased for any of three reasons. First, an increase in the number of individuals at the interface was noticed at low tide. Since it was calm, it is possible that increased pool temp¬ erature during the day induced them to go to the interface to cool off, i.e. air conditioner hypothesis. This would 14 Vertical Distribution of L. scutulata Merchant, Noroian heavily weight the number in the Out plus Interface category. Second, the population was greatly disturbed by marking them. Removal or perhaps nail polish itself could have induced abnormal behavior or triggered various general responses to disturbance, e.g. movement to cracks for protection. Third, by significantly reducing the total population in Pools 2 and 4, intraspecific interactions may have been disrupted, resulting in a change of behavior. It is also possible that the bias lies with the first study. During the last study, migration was seen to occur quickly. Short-term fluctuations in population size in a given pool due to migration can influence vertical distribu¬ tion. For example, Pool 4 is connected by water to another pool, which can act as a source of In Pool snails. It would not be necessary for the snails to move into Pool 4 from the outside; they could just enter without leaving the water. By not observing a constant population in one pool, migration effects may introduce large errors. One of the most interesting aspects of monitoring indiv- idual movement is the sequence of movements observed. There are two important things to be noted in the plots of the numbered individuals. In general, they exhibit the same movement pattern as the total population. This is seen best in Pool 2. However, not all individuals conform; there is a rhythmic cycle which the population follows but at the same time there are individual fluctuations depend- ent upon the recent history of a particular individual. Vertical Distribution of L. scutulata Merchant, Noroian Summary 1.) Fluctuations in the vertical distribution of Littorina scutulata in tide pools were observed at Muscle Point. Pacific Grove, California. 2.) L. scutulata appear to move into and out of tide pools as a function of the tidal cycle. This behavior is modified by the day-night cycle. 3.) 4.) The outward movement is positively correlated with dampness and negatively correlated with wave shock. The actual pattern of the in-out migration in each pool 5.) is complex and dependent on a large number of environ¬ mental variables which are not yet fully understood. The reasons for the in-out migration are still not known. 6.) There are some indications: Movement into the pool and also into cracks seems to offer protection against wave shock; movement to the interface during the day may be a mechanism to regulate temperature. The night¬ time migration from in pool areas, where algae are abundant, to out of pool areas, where algae appear scarce, is not easily explicable. Individual behavior shows a general agreement with 7.) gross population behavior, but deviations were noted. 16 Vertical Distribution of L. scutulata Merchant, Noroian References Glynn, P.W. 1965. Community, Composition, Structure...of the Endocladia-Balanus Association in Monterey Bay, California. Beaufortia 12: 1-198. Hewatt, W.G. 1937. Ecological Studies on Selected Intertidal Communities of Monterey Bay, California. Amer. Midl. Nat. 18: 161-206. Kops, E. 1964. The Effect of Certain Environmental Factors on the Activity Pattern of Littorina planaxis and L. scutulata. Hopkins Marine Station Spring Course Report. North, W.J. 1954. Size, Distribution, Erosive Activities and Gross Metabolic Efficiencies of the Marine Intertidal Snails, Litto prina lanaxis and Littorina scutulata. Biol. Bull. 106: 185-197. Peterson, R.E. 1964. Lower Limits of the Habitats of Littorina scutulata and Littorina planaxis. Hopkins Marine Station Spring Course Report. 17 Vertical Distribution of L. scutulata Merchant, Noroian Acknowledgment We wish to thank the faculty and staff of Hopkins Marine Station for making this endeavor worthwhile. We extend special thanks to Dr. John Phillips for his incredible patience, imagination and advice which helped us in playing the sorts of games we wanted to play this quarter. 18 e Vertical Dist tribution of scutulata Merchant, Noroian Figure 1. Photographs of Area I and its pools. Pool 1 Is the lower of the two. The maximum dimensions of Pool 1 are 85 x 20 cm with a depth of 8 cm. The maximum dimen¬ sions of Pool 2 are 35 x 20 cm with a depth of 3 cm. Vertical Distribution of L. scutulata Merchant, Noroian 20 Vertical Dis ribution of L. scutula Merchant, Noroian Figure Photographs of Area II and its pools. Pool 3 is on the left in the bottom picture. The maximum dimen¬ sions of Pool 3 are 45 x 22 cm with a depth of 5 cm. The maximum dimensions of Pool 4 are 50 x 30 cm with a depth of 7 cm. Vertical Distribution of L. scutulata Merchant, Noroian Vertical Distribution of L. scutulata Merchant, Noroian Figure 3. Change in the total population and percent of the population Out of Pool during the observation period April 28-30, 1978. Tidal and diel cycles are also indicated. Changes in the total population are indicated by a dashed line. Changes in the percent Out of Pool population are indicated by a solid line. Dotted line indicates areas of uncertainty in percent Out of Pool. Vertical Distribution of L. scutulata Merchant, Noroian tide Pool1 60- 40- 20- 0 80- W Po 2 60- 40- 20- 1 80- Pool 3 60 § 40 20 80- Pool 4 60 40- V 20- tide, 1700 000 0100 o900 o900 1700 0100 1700 -160 -120 -80 -40 -160 -120 80 -40 -0 160 -120 -80 -40 -0 -160 120 -80 —40 Vertical Distribution of L. scutula Merchant, Noroian Figure 4. Change in the total population and percent of the population Out of Pool during the observation period May 16-18, 1978. Tidal and diel cycles are also indicated. The amount of splash and wash is indicated by 's. One * for light and two for strong. Changes in the total population are indicated by a dashed line. Changes in the percent Out of Pool population are indicated by a solid line. tide 80- 60- 40- 20- P 80- 60- 40- + 20 7 0. 8 80. 60- 40- 20 80 60- 40 20 tide (ft.) 1500 Vertical Distribution of L. scutulata Merchant, Noroian Po01 P X Pool 2 1 * X PooI 3 * * * * +4 * Pool 4 500 2300 -160 -120 -80 -40 -160 -120 -80 -40 -160 -120 -80 -40 -160 -120 -80 -40 26 Vertical Distribution of L. scutulata Merchant, Noroian Figure 5. Total numbers of L. scutulata In Pool and Out of Pool for Pools 1-4 for the two periods of observation. Solid lines represent Out of Pool. Dashed lines represent In Pool. Tidal and diel cycles are also indicated. Vertical Distribution of L. scutulata Merchant, Noroian tide April 28-30 240 200+ 160 Number 120- of Snails 80- A0- 0100 0000 1/00 00 1ino ot 5+ tide May 16-18 280- 240- 200- Number 160- Snails 120- 80- 40— 1500 2300 0700 1500 2300 500 2300 0700 Time of Day 28 Vertical Dist tribution of L. scutulata Merchant, Noroian Figure 6. Change in the Out plus Interface population during the observation period June 1-3, 1978. Tidal and diel cycles are also indicated. The amount of splash and wash is indicated by *'s. One * for light and two for strong. Vertical Distribution of L. scutulata Merchant, Noroian tide 25- Pool 2 20 Number 15- of Snails 10- Pool 4 20- Number 15- Snails 10— 5- 1700 ojoo o9oo 1700 0100 o900 1700 0100 Time of Day Vertical Distribution of L. scutulata Merchant, Noroian Figure 7. Change in the Interface population during the observation period June 1-3, 1978. Tidal and diel cycles are also indicated. The amount of splash and wash is indicated by *'s. One * for light and two for strong. tide number snails number snails Vertical Distribution of L. scutulata Merchant, Noroian 4 l6- 14- Pool 2 12- j0- 8 - 6- A- 2- L 0 + Pool 4 tta 1700 0100 0900 1700 0100 Time of Day * * 0900 1700 0100 Vertical Distribution of L. scutulata Merchant, Noroian Figure 8. Change in the population in cracks during the observation period June 1-3, 1978. Tidal and diel cycles are also indicated. Changes in the crack population in Pool 2 are indicated by a solid line. Changes in the crack population in Pool 4 are indicated by a dashed line. Vertical Distribution of L. scutulata Merchant, Noroian 4 tide, 24- 22- 20 18- 16- Number of Snails 12- 10- 8 - 4— 2- 10o ol00 0000 Vo ol00 000 100 Time of Day o100 Vertical Distribution of L. scutulata Merchant, Noroian Figure 9. Change in the Bottom and Side populations in Pool 4 during the observation period June 1-3, 1978. Tidal and diel cycles are also indicated. The amount of splash and wash is indicated by *'s. One* for light and two for strong. Changes in the Bottom population are indicated by a solid line. Changes in the Side population are indicated by a dashed line. tide Number Snails Vertical Distribution of L. scutulata Merchant, Noroian * 14— 12- 24 10- 6- 2— L 1700 o1oo 0900 1700 o10o o900 1700 0100 Time of Day 36 Vertical Distribution of L. scutulata Merchant, Noroian Figure 10. Change in location of 8 numbered individuals at Pool 2 during the observation period June 1-3, 1978. Tidal and diel cycles are also indicated. Categories are identified as follows: O indicates Out; I indicates Interface; P indicates Pool. Vertical Distribution of L. scutulata Merchant, Noroian tide p — — kkll P nO- 9 P p P P Voo oloo o/00 100 1700 0100 o900 Time of Day 000 Vertical Distribution of L. scutulata Merchant, Noroian Figure 11. Change in location of 14 numbered individuals at Pool 4 during the observation period June 1-3, 1978. Tidal and diel cycles are also indicated. Categories are identified as follows: O indicates Out; I indicates Interface; S indicates Side; B indicates Bottom. Vertical Distribution of L. scutulata Merchant, Noroian - -p 0-o r slr lr p 1121 2 1P2 40