LABORATORY OBSERVATIONS ON THE NOCTURNAL VERTICAL MIGRATION BEHAVIOR OF TEGULA PUL. G0 (GASTROPODA, TROCHIDAE) Charles J. Wright Hopkins Marine Station Mussel Point Pacific Grove, CA INTRODUCTION Tegula pulligo is the commonest trochid gastropod found in the kelp forests of Monterey County. Previous distributional studies on T. pulligo, T. montereyi and T. brunnea by Lowry and Pearse (1974), Hunt (1977) and Sellers (1977) indicated that the three populations stratified on Macrocystis plants, with T. pulligo usually occupying the lower four meters of the plant. Preliminary subtidal observations in the kelp forest adjacent to the Hopkins Marine Station at Mussel Point in mid-April yielded data suggesting that T. pulligo underwent a diel vertical migration on the kelp, with up- ward motion occurring at night. Subsequent field studies by Thornton (reported in this volume) have shown there to be a statistically significant migration of the population in our study area. Laboratory studies were undertaken in attempt to 1) demonstrate the observed field behavior in a laboratory environment and 2) to determine what effect light, or its intensities, may have on any observed behavior. If light was shown to be an important regulating factor, it was also hoped to obtain a general light-preferred range and maximally tolerated intensity. MATERIALS AND METHODS All experiments were conducted using a clear acrylic cylinder measur¬ ing 2 m high and having an internal diameter of 14 cm. A single kelp plant, usually having between eight and twelve stipes, and its holdfast, was attached to a weighted plastic box and sunk in the cylinder. The box was perforated and prevented animals from blocking water flow, and decreased the directional nature of the flow by allowing multiple outlets or inlets, depending on flow direction. Blades on each plant were trimmed as necessary (2) to allow viewing inside the cylinder. Animals collected at random from the field were dropped in the cylinder and given between one and three days to adjust to their new environment before observational periods began. During several studies, natural light passing through a skylight above the apparatus was the sole light source. In those studies which used artificial lights, 8 75 watt GE reflector bulbs and 1 650 watt Sylvania DWV movie light were used. To allow additional flexibility in establishing desired gradients, 4 of the 75W bulbs and the 650W bulb were controlled by a Variac regulator, and varying thicknesses of window screening were attached to the outside of the cylinder. All light intensities reported were measured with a Weston Model 703-60 Type 3A photometer on the outside of the cylinder. A red-cellophane-covered flashlight with a peak intensity of 50 Foot-Candles was used to determine the position of the snails at night. Fresh sea water (about 14°C) flowed continually throughout the system. Unless otherwise noted, all statistical analyses were done by Student's T-test. RESULT! Figure 1 is a schematic presentation of the light intensities generally encountered in the field. As the study progressed, the canopy thickened, and ambient intensities decreased slightly. Figure 2 presents the results from a study using natural daylight and one day of acclimation. It shows a slight tendency for the animals to move up in darkness, but statistical tests between the mean heights during the dark interval and those of the lighted periods show no signifi¬ cance (p».9). Additional analysis between bright (200 ft.-candles) and dimmer conditions heights did not show any significance to exist, either (.5 «p «.9). (3) The animals represented by Figure 3 were allowed one day of acclima¬ tion and artificial light was used. There appears to be vertical movement with peak height occurring in the brightest light interval. Testing be¬ tween the mean heights of the light versus dark periods showed a close to, but not quite (p«.065) significant displacement of the animals with differ¬ ing light conditions. Further, no significance between dim and bright conditions was demonstrated (.2 «p «.4). In the experiment represented by Figure 4, the lighting was the same same as Figure 3 but 4 days of acclimation was used. The population ex¬ hibited a clear migration upward corresponding with the dark interval. The significance of the movement was dramatic (po.0001). Again, tests were conducted between bright and dim mean heights, and they showed no significant differences to exist (.2 «p«.4). The data depicted on Figure 5 came from a study where dim artificial light was used (a peak of 70 ft. candles) and one day was allowed for acclimation. The mean height during the lighted intervals is rather significantly lower (0.02 « p « 0.05) than the mean heights during the dark condition. Figure 6 presents the data from a displacement experiment. Animals were removed, then dropped back into the cylinder in light. They were given 6 hours of light followed by 9 hours of dark. One run showed a highly significant (using 952 confidence intervals established by T-test) increase in mean heights during the dark interval. The second run showed a dramatic increase also, but it occurred during the lighted period. In combining the two runs, however, no significant differences between dark and light heights can be shown (.05 « p «.1). (4) Figure 7 depicts displacement studies during the night. In analyz¬ ing the first run, there was no significance evident between the light and dark conditions. During the second run, however, highly significant movement took place between the dark period of 1830-1000 and the following lighted interval (p « .005), and motion significant to between .025 and .Ol occurred during the dim period between 1700 and 1830. DISCUSSION As mentioned in the introduction, T. pulligo has been shown by field observations to undergo a significant vertical migration up kelp plants at night, with downward movement as daylight increases. Predation by visually-cued animals known to prey on T. pulligo such as Enhydra lutris, Hexagrammacus decagrammus, Damalychthys vacca and Octopus (Hunt, 1977) could make such behavior selectively advantageous. During the day, though some individuals may be found exposed on the blades and stipes, the bulk of the population remains low on the plant, hidden from view, thereby escaping detection by many predators. At night, a larger percentage of the population moves up onto the stipes and blades feeding. Such behavior, however, was not clearly and consistently seen in the laboratory. With the notable exception of the population of animals used for the Figure 5 experiment, those animals given one day or less to acclimatize to the system showed no significant movement of mean height with varying light intensities. Animals which had been in the system for two days or more began to show movement bordering on significance (p «.05). The animals allowed four days or more in the system before being observed demonstrated a clear migration of population mean height as a function of light intensities. Their peak height was attained during the dark intervals, as was the peak of the field population. This would indicate that the animals required a longer period of adjustment to the experimental conditions than had been anticipated. The data presented on Figure 5 also calls to attention the importance of accurately determining the light intensities actually seen by the animals, both in the field and within the apparatus. As seen in Figure 1, the light experienced by the animals on the kelp appears to be in the 0-125 ft.-candles range. Only during the study presented by Figure 5 was the light that dim; in the other studies, the light shown on the animals tended to be higher for at least a part of the study. The longer period of adjustment could be in response to those higher levels of light. Determining the actual levels seen by the animals within the cylinder was also difficult, since intensity varied as readings were taken around the circumference. In future work, it might prove informative if distinctions were made as to the location of an individual, i.e., behind a blade, in the center of the cylinder, on an extreme wall. The variability of responses within the laboratory system could be a reflection of not having taken such considerations fully into account. Individual variability was touched upon only superficially by tagging and monitoring individual's positions, and could use more work. Samples of the field population were haphazardly taken, and their treatment was not standardized. Perhaps by controlling for such specific factors as sex, size, time since last feeding, or collection site variability of response might be reduced. SUMMARY AND CONCLUSTONS 1) T. pulligo undergoes a nocturnal migration in the laboratory similar to that seen in field studies. (6) 2) T. pulligo requires at least three days to acclimatize to a laboratory environment in which light intensities are higher than those experienced in the field. 3) Animals knocked off plants undergo their greatest repopulation move- ment at night. LITERATURE CITED HUNT, D. E., 1977. Population Dynamics of Tegula and Calliostoma in Carmel Bay, with Special Reference to Kelp Harvesting. M.A. Thesis, San Francisco State University, San Francisco, CA. 81 pages. LOWRY, L., A. MCElroy and J. Pearse, 1974. Sublittoral Ecology of Kelp Beds of the Open Coast Area Near Carmel, California. Biol. Bull. 122: 95-114. SELLERS, R. G., Jr., 1977. The Diets of Four Species of Calliostoma (GASTROPODA, TROCHIDAE) and Some As¬ pects of Their Distribution Within a Kelp Bed. M.S. Thesis, Stanford University, Stanford, CA. ACKNOWLEDGMENTS I should like to recognize the people whose assistance and help in the writing of this paper was indispensable. The critiques and suggestions offered by my advisor, Dr. Chuck Baxter proved timely and very valuable. The as¬ sistance and comments of Dr. John Phillips also are gratefully acknowledged. Dr.Robin Burnett's help in the statistical analyses of the data are greatly appreciated. I would like to thank Sharon for having done the final typing of the paper. And Kath's support and patience have been absolutely astounding, and I can't thank her enough. FIGURE 1 Represented on Figure 1 is are the general light levels found in the kelp forest off Hopkins Marine Station. Intensities were measured between May 9 - 26, 1978. (10) 10 M OPEN WATER 0600 1500 750+ 75-100 500-700 20-35 300-500 10-20 100-250 5-15 Sunny Day 10-15 Ft. Visibility Slight Surge Some Particulate in Suspension 50-100 0-10 Figure 1 (11) so 10050 Sporophy O-10 KELP BED ANO 100-175 FIGURE 2 Animals were allowed one day to acclimatize under natural light. Light intensities were measured at the 1.5 M and 0.5 M levels on the outside of the system. In the lower figure, the dashed line indicates the mean height of the entire population; the solid line depicts the mean height of the animals found above the top of the plastic box to which the plant was attached. The top figure plots the distribution of snails as per¬ centages per quarter cylinder interval. The numbers within the bottom interval refer to the percentage of the total population located below the top of the supporting box. (12) 0. 5 50 25 50.0 200- 90 TIME 56.9 200- 28 —18 49. 38.6 50.456 345 8-0 28.9 130- 18 23 9o- 45.5 750+- 200 12 55.2 (13) FIGURE 3 The group used in this experiment were given one day within the system prior to observations under artificial light. The dashed line is the mean height plot for the total population; the solid line is that of those animals above the top of the supporting plastic box. The top figure plots the distribution of animals as percentages per quarter cylinder intervals. The numbers within the bottom intervals refer to the percentage of the total population found below the top of the supporting box. 120 90 37.7 352 — — — —— 750+ - 70 TIME 20.7 18 450- 30 25.9 27.5 41.3 44.3 526 450-30 De 14 31.7 (15) FIGURE 4 Animals were run simultaneously with those of Figure 3, but had different history. Given one day initially to adjust, the animals were used in a two day displacement study, which involved being displaced four times during the study. Thereafter, the animals were left undisturbed in the same conditions as Figure 3 group. However, at 1600 on the first day of this study, the animals were displaced in a move necessary to standard lighting seen by both groups. Light intensities were measured between the two systems, and are in foot-candles. The dashed line is the mean height curve for the entire population; the solid line is the plot of mean height for those animals above the top of the plastic supporting box. The top figure shows distribution of the total population as percentages per quarter cylinder intervals. The numbers within the lowest intervals are the percentages of the total population found below the top of the box. (16) 120 60 30 55.1 750+- 70 4.7 450- 30 18 TIME 12.4 14.6 — —— ure 31.2 52.7 450-30 513 7504-70 1 5.1 41.9 (17) FIGURE 5 Animals were given one day to adjust under artifi¬ cial light. Peak intensity during the study was 70 foot candles; the cross-hatched interval indicates a slight light leak. The lower figure follows the mean height of the animals above the top of the plastic box supporting the plant. The upper figure shows the distribution of the animals as percentages within quarter cylinder intervals. The numbers adjacent to the lowest interval indicates the percentage of the total population located below the top of the plastic box. 75 50 64.5 57.6 31-0 51.4 50.0 47.2 55. 46.9 54.1 58.3 66-Oft. candles 18 TIME 00 Figure 5 06 12 (19) FIGURE 6 Graphed is the data from a day displacement experi¬ ment. Animals were given one day under artifical lights to adjust. They were then removed from the cylinder, and dropped back in along the walls, to simulate being knocked from plants by surge during the day. A period of six hours light was followed by nine hours of dark. The solid line traces the mean heights for the first day; the dashed line follows the mean heights for the same group on the second day, after a simulated night displacement experiment. 105 35 70 +- TIME 18 ure 06 (21) 750+ - 40 2 FIGURE 7 Figure 7 presents the data from a simulated night displacement experiment. The same animals as were used for the Figure 6 experiment were removed and returned to the cylinder in darkness. After six hours of darkness, lights were turned on. The lower light regime plot indicates the times when a light burned out, dimming ambient levels until a replacement bulb could be obtained. The solid line indicates the mean heights during the first day; the dashed line outlines the mean height curve for the second day. As no distinction between above or below the box top were made, mean heights are for the entire population. 105 7504 40 H I 750+ - 40 12 TIME 15 —11— Figure 7 12 (23)