N. paleacea on P. torreyi. Gansel, J. A. INTRODUCTION The geographic range of the Pacific North American surfgrass, Phyllospadix torreyi (Wats) extends from Oregon doun to Baja California (Munz and Keck, 1963). A root system consisting of rhizoids anchors to rocks and coarse sands in the lower intertidal regions. This position in the intertidal provides a buffer against desiccation and exposure. Air pockets permeate the P. torreyi blades possibly increasing internal photosynthetic capacity (Barbour and Radosevich, 1979). Additionally, these air pockets provide the necessary bouyancy to suspend the blades as a dynamic forest and prevent entanglement with other bottom masses. Beneath the water's surface and suspended above the bottom, the surfgrass blade community offers a reasonably well protected habitat to any lightweight organism capable of holding on. The chaffy limpet, Jotoacmea paleacea (Gould), is one archeogastropod that seems to have responded to this challenge particularly well. Several major modifications leave N. paleacea ideally adapted to Phyllospadix species. Conforming in shell width to the width of the blade, N. paleacea can withdraw all body tissues into its shell to effectively grasp the blade (lab and field observations). A notch along the right side of the nuchal cavity coupled with rapid ciliary currents allows water circulation and waste removal without relinguishing that grasp (Yonge, 1962). Even defense mechanisms seem centered on remaining in contact with the blades, Where other small gastropods display mushrooming and abandoning behavior to avoid predation, I. paleacea simply clamps down tighter and relies on chemical camouflaging (Fishlyn and Phillips, 1980). Rounding out its specificity to Phyllospadix species, 1. paleacea also relies on the blades as a food source. Parasitism seems to best describe the symbiotic relationship between Gansel, J. A. I. paleacea on P. torreyi Notoacmea paleacea and Phyllospadix torreyi. N. paleacea grazes on the chlorophyl containing epidermal cell layer of the plant, presumably lowering the photosynthetic capacity and decreasing blade strength. In the field, this should result in lessened blade growth rates and decreased resistance to blade breakage. Aggressive feeding by N. paleacea could seriously suppress P. torreyi populations. That N. paleacea does not destroy the P. torreyi populations seems to suggest that some mechanism exists to control N. paleacea grazing behavior. Contained within are the results of a study on the symbiotic relationship between Notoacmea paleacea and Phyllospadix torreyi. Along with a distribution relating N. paleacea body size to exposure, this paper examines residency rates on P. torreyi, gut analyses, rates of blade consumption, and the effects of grazing on photosynthetic capacities of P. torreyi METHODS AND RESULTS Field Studies To determine a distribution based on degree of exposure to wave turbulence and desiccation, Notoacmea paleacea were collected from three sites on the Monterey Peninsula. In order of increasing exposure these sites include: Bird Rock Channel - Hopkins Marine Station, Stillwater Cove - Pebble Beach, and West Beach - II.M.S. Bird Rock Channel refers to the sand and rock bottomed channel running between Bird Rock and shore. At low minus tides this channel transforms into a large tide pool with the majority of P. torreyi blades remaining submerged. At high tide, the area is totally submerged but strong wave action is broken up by the offshore reef. Stillwater Cove lies behind a barrier of offshore rocks which create Gansel, J. A. II. paleacea on P. torreyi a natural breakwater. At low minus tides the collection site consists of numerous shallow tide pools which are succeptable to dessication. At high tides, the area is submerged and strong surf is again broken up. Hest Beach has very little protection against strong wave action. At low minus tides the collection site becomes a channel washed by broken waves. At high tide, direct wave contact results in a relatively high energy environment. Distribution data is presented in figure 1. This data suggests that N. paleacea body sizes are a function of their environment. Statistically, (student "T"-test) all mean lengths and mean heights are significantly different (p £ 0.01) except for length comparisons of Stillwater Cove and West Beach (0.1 2 p 20.05) and height comparisons of Bird Rock Channel and West Beach (0.2 2 p 20.1). Statistics suggest that the populations are overall significantly different in size. This difference corresponds to the differences observed in wave and desiccative exposure. Determination of Notoacmea paleacea residency times involved selectively marking blades of Phyllospadix torreyi with tagged wires. Observations over a seven day period regarding residency were made on 25 blades chosen for their initial habitation by N. paleacea. Approximate shell size was also recorded to ensure that the same N. paleacea were being observed from day to day. All observations for distribution and field studies took place during spring at low minus tides. Collection samples included both the residing N. paleacea and the blade occupied. Data from the blade residency time study appears in figure 2. All 25 blades were occupied by a separate Notoacmea paleacea on day 1 but by day 2 only 84% (21 of 25) remained occupied. Blade number one represents a limpet which joined the study after day one. Blades were chosen because the were occupied by N. paleacea. It was assumed these limpets Gansel, J. A. I. paleacea on P. torreyi represent a random sample with respect to previous time spent on a blade. Statistically the mean blade residency time for first day occupied blades is expected to equal half the actual mean residency time. Calculated from a mean blade residency time for first day occupied blades of 2.88 days, the actual mean blade residency time for N. paleacea on Phyllospadix torreyi is 5.76 days. This analysis is supported by blade number one (figure 2). Offering a known beginning and ending time of blade occupancy, its blade residency time of five days corresponds closely with derived mean blade residency of 5.76 days. N. paleacea also reappeared on abandoned P. torreyi blades. The extent of this reoccurrence was always limited to one observation period. Rates of consumption of Phyllospadix torreyi by Notoacmea paleacea were determined in an outside aquarium under natural sunlight in unfiltered running sea water at 15 + 1° C. Twelve ungrazed blades, each approximately 25 cm long, were suspended on a wooden rack in order to keep them separated. Collected N. paleacea were transfered to these blades. Lengths and locations of grazed patches were recorded over the following three day period. After observations were completed, two 20 cm portions of grazed and ungrazed blades were dryed overnight and weighed. Data for consumption rates of Phyllospadix torreyi is presented in figure 3. Of the initial 12 Notoacmea paleacea, only 8 survived through the third day. The total length of P. torreyi eaten per day is X - 24.42 mm (S.D. - 19.69) while the length eaten per day considering only the third day is X - 38.63 mm (S.D. - 14.45). M. paleacea were considered dead when they dropped from their blades and no jonger exhibited the ability to reattach. Grazed patches were easy to see and measure. After the three day observation period, the blades were collected, and dryed. The difference between 20 cm of grazed plant (0.0324 g) and 20 cm of Il. paleacea on P. torrey Gansel, J. A. ungrazed plant (0.0412 g) gives a value of 0.044 mg per mm of P. torreyi grazed. Calculations show that 24.42 mm of blade grazed per day corresponds to a consumption rate of about 1 mg of blade per day or considering the last day alone, 1.7 mg of blade grazed per day. This gain corresponds to approximately 1/50 of the wet body weight of an average sized N. paleacea (8.1 mm, 0.083 g). Gut samples were analyzed from freshly collected Notoacmea paleacea. Dissections involved removal of the dorsal half of the shell including the apex. After the stomach was located and slit open, its content was removed and transfered to a slide, where it was spread, and finally mounted in a glucose solution for examination under the compound microscope. In addition to gut analyses, thin slice cross sections of Phyllospadix torrevi grazed portions were also mounted and examined under the compound microscope. The mounted Phyllospadix torreyi cross sections revealed grazing of the outer epidermal layer with little or no damage to inner mesophyl layers. Viewed dorsally and anterior, the stomach was located just to the right and below the intestine. Its contents were dark green with a few scattered reflective grains intermixed. Under the compound microscope, the majority of the stomach contents (approximately 85%) consisted of P. torreyi cortical cell fragments best characterized by parallel rows of single layered cells. Some of these fragments still contained chloroplasts. Far less common (approximately 102), were the two diatom species found. Finally, in small amounts, fragmented xylem cells were observed. These cells were identified by the existence of accompanying pit cells. l The effects of grazing on the photosynthetic rate of Phyllospadix torreyi were measured on a Gilson respirometer (IGRP-14). Paired samples composed of equal lengths of grazed and ungrazed P. torreyi were measured N. paleacea on P. torreyi Gansel, J. A. for photosynthetic activity. Each sample was placed in a respirometer flask under 10 ml of unfiltered sea water and buffered with 1 ml of a 0.035 M. KHCO, and 0.065 M. NaHcO, solution in the side arm of the flask. The running temperature was set to 15° C., the shaking motor to 5, and light at the blades was measured at 247 microeinsteins / me : sec. After allowing for a 20 minute equilibration period, observations were recorded every 15 minutes for 135 minutes. At the end of the observation period, the blades were recovered, dryed, and weighed. After dividing by dry weights, photosynthetic capacity was graphed in figure 4. Choosing a common period of relatively constant 0, production, a linear regression was performed over the 30 to 90 minute interval. Moticably lower slopes indicate a lower photosynthetic rate for grazed plants. Statistically, the difference between grazed (x - 34.05 ul 09 / g : min, S.D. - 4.02) and ungrazed (50.51 ul 02 / g : min, S.D. - 1.96) blades is significant (student "T"-test, 0.05 p 0.02). DISCUSSION The distribution data in figure 1 suggests that environmental conditions such as wave intensity and exposure to desiccation piay an important role in determining the overall body size of Notoacmea paleacea. As environmental strains increase, N. paleacea size decreases significantly. The strong wave action at West Beach could be either selectively destroying larger N. paleacea by selecting against increased drag (increasing size), or acting against the whole population simply by limiting the average age. Decreased size at Pebble Beach could be a function of decreased opportunity grazing activity or some metabolic burden limiting growth, or again, the hostile environment might simply be lowering the probability of survivingto reach larger sizes. Gansel, J. A. N. paleacea on P. torreyi That 4 of 12 Notoacmea paleacea (25%) died in the consumption rate study is probably the result of injuries inflicted during the transfer process from field to lab Phyllospadix torrevi. Considering a chance of injury to the surviving N. paleacea, it seems justifiable to interpret figure 3 by considering day three alone rather than all three days in determining a mean consumption rate. Day three rates have both a greater mean (consumed length per day of 38.63 mm compared to 24.42 mm) and a lower standard deviation (14.45 compared to 19.69). By the third day, N. paleacea seems to be relatively stable on the lab blades. From third day data, N. paleacea consumes approximately 1/50 of its own body weight per day in dry weight of P. torreyi epidermal cells. More significantly, over a 5.76 day period (mean residency period) this amounts to a grazing length of 220 mm. The majority of Phyllospadix torrevi chlorophyl is contained within the epidermal cell layer. Grazing by Notoacmea paleacea upon the epidermal cells would be expected to lower photosynthetic capacity. Photorespirometer results verify this conclusion. Oxygen production is over 30% lower in grazed blades, indication significantly reduced photosynthetic capacities. Overgrazing could have a devastating effect on plant growth with such high rates of photosynthetic loss. The discovery of xylem cells in gut samples indicates that Notoacmea paleacea does at least some grazing much deeper than was previously recorded (Barbour, 1979; Fishlyn, 1980). Grazing deeper than -the epidermal cell layer should seriously weaken the blade; however, this weakening was not observed. Deep grazing then probably does not occur often or for long stretches. In any case, deeper grazing would surely amplify the risks involved in overgrazing. Overgrazing certainly threatens the stability of the limpet - surfgrass N. paleacea on P. torreyi Gansel, J. A. relationship. Without controls on the grazing rates of Notoacmea paleacea, irreparable damage to the P. torreyi population is inevitable. Evidence towards the existence of such a limiting mechanism is found in the residency period study (figure 2). Residency periods seem to be of two types: a full period of residence lasting an average of 5.76 days and an abbreviated residence time lasting a maximum of one day (actual abbreviated period is unknown). Itiis suggested that full residency (5.76 days) indicates an average period the blade is occupied for grazing while abbreviated residency indicates that the blade was found unsuitable for grazing. If this is the case, residence on blades by N. paleacea is not random, but involves a selective process. Perhaps N. paleacea can recognize recent residency on a blade. The suggested selection against previously occupied blades would effectively control overgrazing. Such a mechanism limiting overgrazing is clearly advantagous to both Phyllospadix torreyi and Notoacmea paleacea. With the extreme effects of grazing on photosynthetic capacities, overgrazing would be a serious detriment to the blades. A mechanism limiting grazing ensures that the P. torreyi population will not be irreparably damaged. Notoacmea in turn is dependant upon the prosperity of its host Phyllospadix torreyi. N. paleacea on P. torreyi Gansel, J. A. SUMMARY 1. Distribution studies indicate that differences in Notoacmea paleacea population body sizes correspond to factors of wave intensity and desiccation from exposure to air. 2. The mean grazing rate on Phyllospadix torrevi is 38.63 mm of blade length per day or approximately 220 mm over an average grazing residency of 5.67 days. N. paleacea consumes about 1.7 mg per day dry weight of P. torreyi. 3. Grazing results in over a 302 loss in photosynthetic capacity. 4. Gut analysis shows P torreyi epidermal cells constitute about 85% of the diet of N. paleacea with small amounts of xylem cells and diatoms making up the remainder. 5. Periods of residency of individual limpets on blades of Phyllospadix torrevi may be grouped into two categories: full residency periods of 5.76 days and abbreviated residency periods of one observation. 6 Full residency periods indicate grazing behavior while abbreviated residency periods indicate rejection of the blade as a food source. This points to the existence of a mechanism that enables N. paleacea to detect previous residency. This mechanism controls overgrazing, a control that is critical to the success of P. torreyi and N. paleacea. N. paleacea on P. torreyi Gansel, J. A. ACKNOWLEDGMENTS I wish to extend special thanks to my advisor CHUCK BAXTER for his patience and quidance, to Dr. Isabella Abbott for the trifle and help in analyzing limpet guts, to Robin Burnett for his suggestions on statistical analysis, and to the rest of the Hopkins staff, and fellow students for making the quarter so enjoyable. I can't imagine spending spring anywhere else. 10 N. paleacea on P. torreyi Gansel, J. A. LITERATURE CITED Barbour, Michael G. and Stephen R. Radosevich. 1979. "C Uptake by the marine angiosperm Phyllospadix scouleri. Amer. J. Bot. 66: 301 - 306. Fishlyn, Debby A. and David W. Phillips. February 1980. Chemical camouflaging and behavioral defenses against a predatory seastar by three species of gastropods from the surfgrass Phyllospadix community. Biol. Bull. 158:34 -48. Munz, Phillip A. and David Keck. 1959. A California flora. 1681.pp.; 133 figs.,; Berkeley, Calif. (University of California Press). Yonge, C. M. 1962. Ciliary currents in the mantle cavity of species of Acmea. The Veliger 4: 119 -123. Gansel, J. A. N. paleacea on P. torreyi 12 FIGURE CAPTIONS Figure 1. The relation of length and height of 3 populations of Notoacmea paleacea to increasing levels of exposure. West Beach exposure is greater than Stillwater Cove exposure is greater than Bird Rock Channe! exposure. Figure 2. Residency periods of individual M. paleacea on marked P, tor blades over a seven day period. Figure 3. Lengths of grazed areas on Phyllospadix torrevi blades for individual M. paleacea over a three day period. Figure 4. Photosynthetic activity graphed against time in minutes for grazed and ungrazed blades of P. torrevi. Figure 1 13 BIRD ROCK CHANNEL HMS TTT L STILLWATER COVE PB II T WEST BEACH HMS T L — 6 7 8 9 10 11 3 4 HEIGHT bottom LENGTH top mm. O DAYS BLADE HOLDS A LIMPET N 0 0 14 120 100 80 60 40 . Figure 3 AFTER DAY DAY 2 DAY 3 3 limpet number 15 36.8 o 31.2 ° 2 ao 2 6 8 49.1 4 0 O o 0 0 51.8 0 0 0 2 0 o o 90 120 30 60 minutes Figure 4 16