Brumm-pag Introduction Persistent diurnal rhythms have been discussed extensively by many investigaters (Webb, 1950, Behrnes, 1974). Such rhythms have been observed in both physiological and behavioral processes. (Cloudsley-Thompson, 1961). Tidal rythms, as well as diurnal rhythms, have been observed in intertidal invertebrates (see Korringa, 1947). Sandeen, Stephens and Brown (1956) have noted a persistent diurnal rhythm of oxygen consumption in two intertidal gastropods, Littorina littorea and Urosalpinx cinereus. In Littorina planaxis, two daily periods of greater oxygen consumption were observed (Lebenzon, Hopkins Marine Station, Spring report, 1964) Studies on the general activity of three species of Littorina, L. scutulata, L.sitkana and L. planaxis by Behrens (1974) revealed a marked increase in activity during the hours of darkness. Kops (Hopkins Marine Station, Spring report, 1964) and Haseman (1911) observed similar diurnal fluctuations in the activity of L.planaxis and L. littorea respectively. However, there was no persistence of these rhythms when the animals were placed under labortary conditions. When L. planaxis is suddenly immersed in sea water by wave action, it responds by opening its operculum, extending its foot and attatching to the substratum (Bigler, Hopkins Marine Station, Sprin report, 1964). Preliminary studies revealed variation in the time required for this Brumm-page 3 response. The following is a study of the relationship of response time to phases of the tidal and diurnal cycles. Materials and Methods All snails were collected from the rocky intertidal at the Hopkins Marine Station, Mussel Point, California. Animals were taken from a zone approximately five to six feet above mean lower low water. No attempt was made to select animals from either horizontal or vertical surfaces, but all animals were collected within an area of 2-3 square meters. Response time was measured by dropping the snails into approximately 10 milliliters of sea water contained in 1.5 by 15 cm. test tubes. Care was take to assure that the animals were placed in the water with their opercula up. Samples of twenty-five snails were used and the results expressed as the number of individuals responding within 60 seconds. Tests with five groups of twenty-five snails, collected at high tide and at low tide, gave a standard deviation of 3.85 and 3.42 respectively for the 60 second response time. Results Measurements of the number of snails responding within 60 seconds were mde on groups of snails taken from the intertidal at times corresponding to points in the tidal cycle over a period of 51 hours. Figure 1 shows the variation in the number responding in these Brumm- page 4 sample groups. The mean number responding during the period of observation was 10.9. Only the low number responding at dawn on the first day is significantly different from the rest, due to the high degree of variability in the number responding at any one point in time. However, the fluctuation in the number responding suggests some correlations with aspects of the tidal and diurnal cycles. The difference between the number responding at the peaks of the high tide may be due to a difference in the violence of the waves at the two times. On day I, the waves were significantly larger than on day 2. Figure 2 shows a similar measurement of the number responding during another relatively calm tidal cycle. In this study, the high tide has advanced by four hours and a correla tion between the peak number responding and the high tide is once again to be noted. Although in Figure 1 the increase in the number responding occurs before sunset, this increase occurs well after sunset in Figure 2 suggesting a closercorrelation between the tidal cycle, rather than the diurnal cycle. These field studies suggested a relationship between the number responding and the amount of available moisture in the high intertidal where L.planaxis occurs. The rising tide on relatively calm days provides this area with nonviolent splash. To test the relationship between fluctuation in the number responding and fluctuations in the amount of moisture available, snails were collected at lower, low water, well Brumm- page 5 before dawn when the rocks were still damp, and maintained in the laboratory under conditions of constant moisture and dim light. Constant moisture was provided by placing the snails on wet paper towels. One group of twenty-five were tested immediately. Figure 3 shows the results of this study. The increase in the number responding occurs at a time intermediate between that observed in the studies shown in Fiures 1 and 2 and suggests a possible correlation with the diurnal cycle, as well as the tidal cycle. In a similar experiment, animals were collected at lower low wwater after dawn and maintained in the laboratory under conditions of constant dryness. One-half of these animals were subjected to the constant light of the laboratory. The other half were placed in constant darkness and tested usin only a redlight. The presence or absence of lighthhad little effect during the period of the experiment. Figure 4 shows these results. Although the most striking increases in number responding corresponds to a point in the tidal cycle correlated with high tide and approaching darkness, Figures 1, 3, and 4 also suggest another peak of responsiveness corresponding to the morning hours. Brumm- page 6 Discussion The responsiveness of samples of snails appears to vary during the tidal and diurnal cycles. There is a suggestion from the studies reported here that both of these cycles may influence this aspect of behavior. A separation of these effects was attempted by studying animals maintained under conditions of constant dark or light and constant dryness or moisture. None of these treatments caused a markedälteration in the behavior pattern during the time intervals studied. It appears that studies of longer duration would be required to further ellucidate the influence of these two cycles. However, the number of snails required for these long term experiments would require the use of animals from a much greater area than that selected as a source for these studies. This may introduce an even greater vari¬ ablity in the response of the tested groups. There is a suggestion of two peak times of greatest responsiveness. One occuring in the morning anc the other around sunset. The morning peak was also correlated with lower low water and the evening peak with higher high water in these studies. These two peaks correspond to the times of greatest oxygen consumption, as noted by Lebenzon (Hopkins Marine Station, Student report,1964). Brumm- page Summar 1. The number of Littorina planaxis responding within 60 seconds when immersed in sea water varies during the tidal and diurnal cycles. 2. Maintainence of animals under conditions of constant light, dark, dryness or moisture do not appear to alter the pattern of this variation during periods of up to 24 hours. 3. Two peak periods of greatest responsiveness were observed in samples of snails obtained from the field or maintained as described. One peak period occuned in the morning and the other around dusk. Brumm-page Literature Cited Bigler, E., 1964. Attrition on the Littorina planaxis population. Student report, Hopkins Marine Station, Pacific Grove, California. Behrnes, S., 1974. Ecological interactions of three Littorina (Gastropoda, Prosobranchia)along the west coast of North America, Ph.D. Thesis, University of Oregon. LIl pages. Cloudsley-Thompson, J.L. Rhythmic activity in animal physiology and behavior. Acedemic Press, 1961. New York. 285 pages. Haseman, J.D., 1911. Therhythmical movements of Littor: littorea synchronous with ocean tides. Biol. Bull. 21:113-9 Korringa, P., 1947. Relations between the moon and periodicity in the breeding of marine animals. Ecol. Mono. 17:347-81 Kops, E., 1964. The effect of certain envirnmental factors on the activity pattern of Littorina planaxis and Littorina sctulata. Student report, Hopkins Marine Station, Pacific Grove, California. Lebenzon,J., 1964. Respiration in Littorinia planaxis and Littorina scutulata. Student report, Hopkins Marine Station, Pacific Grove, California. Sandeen, M.I., Stephens,G.C., Brown,F.A.,Jr., 1954. Persistent daily and tidal rhythms of oxygen consumption in two species of marine snails. Physiol Zool. 27:350-56. Webb, H.M., 1950. Diurnal variations of response to light in the fiddler crab, Uca. Physiol. Zool. 23:316-37. Brumm- page Acknowlegements I wish to thank Dr. John Phillips for his patience and helpful advising during the course of the quarter. I would also like to thank Susan Harris for her encouragment at a time when I really needed it. I would also like to thank Linda Baum for her help during the preliminary studies and for her stimulating conversations at 4:00 A.M. 0 Figure 1 Number of L. planaxis responding within 60 seconds. Animals taken from the intertidal at points in the tidal cycle as indicated. Each point represents the number responding in the sample of twenty-five tested. Vertical bars indicate plus and minus 1 standard deviation. — TIDAL CYCLE 18 4 2- L 12 18 12. TIME k — Figure 2 Number of L.planaxis responding within 60 seconds. Animals taken from the intertidal at points in the tidal cycle as indicated. Each point represents the number responding in the total sample of twenty-five tested. Vertical bars indicate plus and minus 1 standard deviation. 2271 2 d 1 11 TIDAL CCLE kv- 0 12 TIME — Figure 3 Number of L. planaxis responding within 60 seconds. Animals were collected from the intertidal at lower low water and maintained in the laboratory under conditions of constant light and constant moisture. Each point represents the number responding in the sample of twenty-five tested. Vertical bars indicate plus and minus 1 standard deviation. 12 S lo¬ N 83 2- TIDAL / t- 12 9 15 kkt - Q 3 21 TME Figure 4 Number of L. planaxis responding within 60 seconds. Animals were collected from the intertidal at lower low water and maintained in the laboratory under conditions of constant dryness. One-half of those collected were kept in constant light, and is represented by the solid lines and dots. The other half was maintained in constant darkness and tested using red light. These are represented by the open circles and the dashed line. Each point for both conditions of light shows the number responding in a sample of twenty-five tested. Vertical bars indicate plus and minus 1 standard deviation. — TIDAU CGE MA K. N L 4 12 15 18 210 3b 9 TIME —