HORIZONTAL AND VERTICAL MOVEMENT IN ADULTS OF THE POLYCHAETE ANNELI CIRRIFORMIA SPIRABRANCHA Philip E. Henderson Hopkins Marine Station Stanford University Pacific Grove, California Philip B. Henderson Cirriformia spirabranche (Woore, 1904) is very common along the California coast, inhabiting mainly sheltered intertidal regions, although at times it extends to depths in excess of 7 meters. During the spring of 1968, preliminary examinations were made on various populations of intertidal C. spirabrancha in the yacht harbor area of Monterey, California. During these observations core samples taken at the 0.0 ft. tidal level at both low and high tide showed the presence of large numbers of animals when the sand was wet, and an apparent complete absence of animals in the same region at extreme low tide. This suggested possible vertical or even horizontal movement during various stages of the tidal cycle. While work haskbeen carried routdonusome aspectspect of the biology of cirratulids by George (1964), Flatterley (1916), and Courtney (1958), there have been no investigations concerning the movement of the adult animals. Because of this general lack of knowledge concerning the movement of adult polychaetes and because initial field observations on C. spirabrancha suggested rather extensive movements of the adults of this species, an investigation was initiated to determine the presence, magnitude and characteristics of movement of the adults of C. spirabrancha Philip E. Henderson MATERIALS AND METHODS Field experiments were conducted in the yacht harbor area of Monterey, California, during the spring of 1968. The directional movement of adult C. spirabrancha in a population at about the 0.0 ft. tidal level was investigated in the following three experiments. To test for horizontal and diagonal movement two 1xlx0.33 ft. wood-framed containers, screened on the top and open at the bottom, and two O.5x0.5x0.33 ft. wood-framed boxes were used (figure 1). A one cubic-foot hole was dug during a very low tide. The large box was placed in the hole with its screen facing downward. One of the smaller boxes was placed at its center. Sand extracted from the hole was sifted into the opening to remove any adult Cirriformia which might have been present. When the sifted sand was level with the surface, a smaller box was placed at the center and forced into the sifted sand. The remaining large frame, screen facing upward, was forced down until the screen touched the sand. Screens allowing for the passage of water were specifically placed to prohibit the worms from entering from above or below, yet still allowing them to enter from the sides. The substrate between the two smaller boxes was examined every third day for a total period of twelve days. To test possible movement through the surf, a 0.125 cubic-foot container with wooden sides, screened bottom, and open top, was buried during a very low tide, its top flush with the substrate surface (figure 2). To prevent worms from entering over the lip, a flanged screen collar was extended Philip E. Henderson from the lip 3 in. outward to cover the surrounding sand. The box was filled with sifted sand taken from the area. The sand in the container was examined 4 times during a 12-day period. To test the range of vertical movement, a 4x20 in. polyethylene cylinder, filled with sifted sand and capped on the top, was buried during a very low tide. The cylinder was removed at high tide and the contents examined for the presence of the polychaetes. This experiment was repeated times over a 12-day period. Preliminary studies indicated that tentacles of some individuals were present on the substrate surface throughout the tidal cycle except when the sand surface was above the water level and dry. Movement patterns of the population were charted by placing dyed animals in given areas and observing their dyed tentacles on the surface of the substrate. Neutral red, Bismark brown, and Toluylene blue were found to be the most effective dyes after tests in which the organisms were submerged for 2 hrs. in a solution containing 1 liter of sea water to 2 gms. of dye. 500 worms were placed at various stations in the field area. The number of worms was measured volumetrically since excessive handling was found to be fatal. Approximately 500 worms can occupy a volume of 300 ml., hence 300 ml. containers filled with dyed animals were estimated as containing 500 animals. Four stations were chosen on the basis of their variability in three pertinent conditions: density of the Philip E. Henderson existing population, sand particle size, and tidal level. Station A was at a depth of -2.5 ft. The substrate was very fine, with 7%% of the sand having a diameter of less than 0.124 mm. (as determined by dry weight). The interphase between aerobic and anaerobic substrates (white vs. black sand) was 4 cm. below the surface. The population density was found to be 3,200 worms/m as determined by random cores. Station B was situated at 10.7 ft. The sand here also was fine, with 76% of it having a diameter less than 0.124 mm. The interphase was 5 cm. deep. The worm population density was 2,700/m. Periodically, mild turbulence resulted from tidal changes. Station C was situated at 40.3 ft. The sand size was much coarser here with 19% of the sand larger than 1.397 mm. in diameter and 45% smaller than 0.124 mm. in diameter. The area was surrounded by large boulders, which might possibly hinder horizontal movement of the worms. The population density was 750 worms/m. Station D was situated at +0.1 ft. It was centered on a broad, sandy plain, where tidal changes created much turbulence. The particle size of the sand was coarse, with 24% larger than 1.397 mm. in diameter and only 29% smaller than O.124 mm. in diameter. Population density was 1,900 worms/m?. Neutral red-dyed worms were placed 3 in. deep in a in. diameter hole (the "origin") att each of the stations. For five Philip E. Henderson days, these areas were observed at the 4 extremes of the tidal cycle. The number of tentacles and the movement of the tagged individuals were recorded. Each station was observed for an additional two days only at high tide. On the 8th day, a 2 ft. diameter hole was excavated at each of these "origins" to determine the number remaining. The experiment was repeated using worms dyed in Bismark brown. The tagged worms were observed during the extremes of the tidal cycle for two days and during high tide for two additional days. The test was again repeated using Toluylene blue to dye the tentacles. The animals were observed at high tide for 6 consecutive days. Afterwards, the sand in a two foot diameter circle around the origin was sifted and the number of dyed individuals recorded. It is extremely difficult to determine the number of tagged individuals present near the surface. Each Cirriformia extends an estimated 5 to 10 tentacles above the substrate. When a population was scasparse, it was assumed that all tentacles projecting from the same point belonged to a single worm. Based on the assumption that each worm put 7 tentacles above the surface, a visual estimate was made of the worms present in a dense population. Tentacles on the surface do not necessarily indicate the density of the population present under the surface of the sand. Consequently, cores were taken at stations B, C, and D during the four extremes of the tidal cycle to determine whether there was a constant ratio of tentacles on the r surface to the worm population density underneath. Tentacle Philip E. Hendersor numbers were recorded in a 4 in. diameter circle and a 10 in, deep core was taken. Since the density of tentacles on the surface is variable, efforts were made to include both "heavily" and "lightly" populated areas as estimated by the number of tentacles apparent on the surface. Meanwhile, an attempt was made to determine whether the same relationships occurred in samples taken during a night- day cycle. Four containers 1 ft. and 5 in. deep were placed in an outdoor tank. The containers were filled with sifted substrate and with 30, 150, 250, and 500 worms respectively. Water height was kept at a constant level of 1 ft. above the sand surface. The ratio of tentacles to the total population was recorded day and night for 4 days. RESULTS AND DISCUSSION Cirriformia appear to move diagonally and possibly horizontally. The submerged cylinder testing the possibility that animals migrate vertically below twenty in. failed to detect migration from these depths (table 1). If any worms had descended below twenty inches during low tide, they should have returned again toethe surface at high tide when tentacles of Cirriformia reappeared in the adjacent area. Observations that neither Cirriformia nor their mucus tubes were present below eight in. (Smith, 1968) likewise confirmed this. Movement through the surf cannot be an important means of transportation; not even an occasional worm appeared in the container which allowed only surface moving organisms to enter, except that in one instance two worms were found in Philip E. Henderso the apparatus. Heavy surf, however, had buried the container under three in. of sand, possibly allowing entrance through the substrate (table 1). The container allowing horizontal and diagonal migration did contain many worms after three days. If the worms entered horizontally from the surrounding substrate, they should have dispersed randomly throughout the container. In fact, Notomastus, an errant polychaete commonly found in the same substrate as Cirriformia, was evenly dispersed throughout the container. If movement were diagonal, however, worms could reach the center box only by a zig-zag pattern of burrowing-- moving upward diagonally, retreating along a different path, and then coming up again in a different direction. Few worms would therefore be expected in that case to reach the inner box. Indeed, a disproportionate number of Cirriformia were found in the area between the small boxes. Only an average of ten percent of the worms in the entire container were in the middle section which occupied thirty percent of the total volume. Tentacle counts indicated that at all times only approximately forty-five percent of the animals had their tentacles showing above the surface of the sand, except when the sand was uncovered by water, in which case all the tentacles were retracted (table 2). There was no regular migratory pattern noticedlduring experiments in which the movement of dyed worms were observed. The dyed worms were dispersed equally in all directions from the four "origins" (table 2). The rate of movement was -9- Philip E. Henderson slow, after sig days a few individuals had moved as far as five feet, but an average of eighty percent of the worms were still clustered about the "origin" (figures 3 and 4). Unfortunately, the staining process seems to have had lethal effects on the organisms. When the "origins" were excavated at the termination of an experiment, no more than half of the worms originally planted could be accounted for and decaying Cirriformia pieces were observed on two occasions. During other experiments, several stained worms were seen protruding from the substrate with decomposing skin. Yet many stained worms lived for over one month with no apparent side effects. The unexplained disappearance of Cirriformia which initially led to this study could perhaps be accounted for by the aggregation tendencies of the worm. Concurrent experiments (Smith, 1968) indicate that the worms often are found in densities greater than 750/100 cm in 1 to 15 foot diameter clumps, 1 to 5 feet apart. Practically no worms were found between such clumps. The initial coring may by chance have included only unpopulated areas during low tide and only aggregated areas during high tide. Results of these experiments, then, indicate little movement of adult Cirriformia. The movement that does occur appears to be small and random. These results suggest that populations of adult C. spirabrancha are relatively stable and tend not to move once established. Philip E. Henderson SUMMAR Movement of Cirriformia is random and not extensive. 2. Cirriformia appear to move diagonally, and possibly horizontally 3. The number of tentacles observed on the surface of the substrate in a given population seems to represent only 35 -55% of the total population during night and day, and during all phases of the tidal cycle except in dry sand during an extremely low tide when their tentacles are retracted. 4. Evidence indicates Cirriformia remain in the upper 8 inches of the substrate. ACKNOWLEDGEMENTS This work was supported ta part by the Undergraduate Research Participation Program of the National Science Foundation Grant GY-4369. My sincerest thanks are given to the faculty and staff of Hopkins Marine Station of Stanford University for allowing me to do this study, and especially to Dr. Welton Lee for his suggestions and guidance in the execution of this paper. I am also gratefully indebted to Mr. Roger Szal for his encouragement and assistance. C Philip E. Henderson REFERENCE! Courtney, W.A.M. 1958 Certain aspects of the biology of the cirratulid polychaetes. Ph.D. Thesis. Univ. of London. Flatterley, F.W. 1916. Notes on the ecology of Cirratulus (audouinia) tentaculata (Montagu). J. Mar. Biol. Ass. U.K., 11: 60-70. George, J.D. 1964. One some evnironmental factors affecting the distribution of Cirriformia tentaculata at Hamble. J. Mar. Bio. Ass. U.K. 44: 383-388. Smith, D.A. 1968. Personal communication. 186 0 ermanent Address FOOTNOTES C Tabl Number of C. spirabrancha found in apparatus testing horizontal and diagonal movement through the substrate (A), vertical movement below 20 in. (B), and movement through the surf. (C). Tria between inner boxes 2. between 63 outer boxes Table 2 Dispersion pattern of Cirriformia. On 6 consecutive days, the number of worms in each quadrant surrounding the "origin" were counted. The table shows the mean percentage and standard deviation as a percentage in each quadrant. Quadrant III Station A 289 20% Mean 26% 26% 5% 8% Standard Deviation 5% Station B 25% Mean 27% 16% 4% Standard Deviation 6% Station C 26% 25% Mean 259 24 5% 7% 6% Standard Deviation Station 25% 29% 26% Mean 4% 3% Standard Deviation Philip E. Henderson Table ? Ratio of the number of Cirriformia showing tentacles on the surface to total population in the same area. Counts were made during two nights and days in the laboratory under conditions of constant water height. The remaining counts were made in the field for three days at 3 tidal extremes. erio No. of samples Mean ratio Standard Deviation Nigh .45 Day .46 High Tide .49 Lower High Tide .43 .11 Higher Low Tide .45 TABLE 4 Number of dyed worms in substrate within 1 ft. radius of the 'origin" the day after completion of the experiment. It was impossible to obtain data from Station A. Initially, 500 worms were present at each station. Experiment I Experiment II Experimer Station B 15. 236 Station C 21. 33. Station FIGURE LEGENDS igure 1. Apparatus testing diagonal and horizontal movement of Cirriformia through the substrate. Figure 2. Apparatus testing possible movement of Cirriformia through the surf. Figure 3. Distance and number of dyed tentacles observed away from the origins 6 days after the third experiment. The organisms within one foot were not included since the majority were in an aggregation within that åreasneEach number"on the abscissa represents the distance from the "origin." The points on the graph represent the total number of dyed animals observed between the corresponding distance from the "origin" and the next recorded distance. Figure 4. Total number of dyed worms at and near the "origins," seen during a series of high tides for 6 consecutive days after implantation. The results are indicative of those obtained in two additional experiments. 20 I — 0 Eu a O 0 m 2 2 2 2 00 0 0 0 d 400 0 —OO NOGC 6 9 10 2 10 1 0 0 2 5 NO. DYED TENTACLES OBSERVE oo888980 O—-o 0 O 0 0 D 1— — OC 2 »