Feeding Differences between Small and Large Individuals of the Sea Anemone Metridium Senile Jennifer E. Purcell Hopkins Marine Station of Stanford University June 7, 1975 Running title: Feeding Differences in Metridium Direct all proofs and correspondence to: Feeding Differences in Metridium Metridium senile is an abundant anemone of circum- polar distribution. On central and southern Californian shores it occurs in large numbers in protected bays and harbors (Hand, 1955). Much of the earlier work on M. senile is summarized in Stephenson (1935), and the num- erous papers of Batham and Pantin (e.g. 1950, 1950) cover many aspects of structure and function. Previous studies include some work on the feeding behavior in Metridium. Parker (1898, 1905, 1917) des- cribed reactions to food, ciliary currents and their reversal on the lips and pharynx, and fatigue in the feeding response due to repeated stimuli, but made no investigation of the natural diet. Stephenson (1935), echoing the opinions of Elmhirst (1925) and Pax (1928), assumed that Metridium are zooplankton-feeders. This conclusion, reached also by Hand (1955), is based solely on the following laboratory and experimental observations. 1) Metridium accepts macroscopic food less readily than other anemones. 2) They live for years in flowing seawater aquaria with no food other than the plankton in the water. 3) Large individuals reject or respond sluggishly to macrofood. Elmhirst noted that large Metridium reacted reluctantly even to copepods and crab larvae in the lab. Small individuals accepted pieces of mussel and fish readily, but seemed not to depend Feeding Differences in Metridium on larger food. Hand attributes this "change in food habits" with body size to the intertidal distribution of the animals; small anemones live near shore where the cresting waves carry larger food to them,, while the larger anemones, living in deeper water receive a constant supply of plankton. No studies have yet been conducted to determine the natural diet of Metridium of any size, or to sys- tematically explore the feeding habits to test whether the feeding preferences of small and large animals in- dicated above are real and consistent differences. This study was undertaken to discover the natural foods of large and small individuals of Metridium. Habitat of the Study Animals The research was conducted at the Hopkins Marine Station of Stanford University in Pacific Grove, Cal- ifornia during April and May 1975. Animals were collected from Monterey Wharf #2. There is a rough distribution in size with smaller anemones tending to be higher on the pilings and closer to shore. The very large Metridium (20-35 cm in column height) occur only at the end of the wharf within a few feet of the bottom. Small individuals (less than 6 cm) intermingle with the Feeding Differences in Metridium intermediate sizes (10-15 cm), and are often densely arranged on the pilings. The largest animals tend to be more solitary. However, all of the smaller and inter- mediate sized specimens used in the present work were collected from areas immediately around large individuals. Morphology of the Tentacular Crown A description of the oral disc and tentacles of the different sized individuals of Metridium is essential to understanding their feeding behavior (Fig. la, b, and c). The smallest animals (less than 3 cm in column height) have an unlobed disc bearing tentacles about 1 cm in length, arranged in only a few rows. From this size through the intermediate sizes the disc becomes larger and progressively more lobed; tentacles remain the same size but many more rows of tentacles are added. There is a hiatus in the continuum of both complexity and size between the intermediate and the large animals. In the largest Metridium the disc is elaborately lobed with six to eight petal-like extensions. The tentacles remain about 1 cm in length but are very numerous es¬ pecially at the margins of the disc and its lobes. Several rows of larger tentacles surround the mouth and extend partially down the center of each lobe. The Feeding Differences in Metridium opposing edges of each disc lobe are usually held fairly close together, forming a solid thicket of tentacles on top and a conical space below between tentacles and disc in each lobe. Feeding Mechanisms in Metridium Reactions to different sizes and types of food were observed in the various sizes of Metridium. Smaller food was dispensed by a Pasteur pipette at a distance such that the prey did not swim away but also was not swept into the tentacles by the force of ejection. Larger foods were presented with forceps. Foods pre- sented included Artemia larvae of two sizes (1 mm and 1.5-2 mm long), Artemia adults (1 cm), the copepod Tigriopus californicus (1.5 mm), and pieces of the squid Loligo opalescens and the mussel Mytilus californianus (1 cme). Responses were observed to food placed on the tentacles at various distances from the mouth of the anemone. The reactions of anemones 3-15 cm in height were similar, and these animals are treated as one group, Newly hatched Artemia larvae (1 mm) were not caught by the small Metridium, but swam among the tentacles and were seen to touch them with no apparent effect on either animal. Slightly larger Artemia larvae (1.5-2 mm) Feeding Differences in Metridium were killed and caught readily with one tentacle, as were Tigriopus of the same size. The method of transfer of food to the mouth differed according to the distance of the food-catching tentacle from the mouth. Tentacles near the mouth carried the prey by ciliary current to the tentacle tip, then bent letting the food fall into the mouth. Tentacles on the edges of the oral disc bent medially until the prey touched another tentacle where it attached. It was then released from the first tentacle and the process repeated until the food reached an inner row of tentacles; ciliary currents then carried it off the tentacle tip into the mouth. Large Metridium responded differently to Artemia larvae and Tigriopus. These foods were always caught and held with one tentacle. The prey remained attached to the tentacle, no movement toward the mouth occurring. Shortly after capture, the anemone closed completely. For small Metridium, the capture of active Artemia adults required the participation of a localized group of tentacles. The portion of the anemone's disc involved in capture increased the larger the prey was in relation to the size of the anemone. After food capture the contracted group of tentacles relaxed and the prey re- mained attached to one tentacle. The prey was then trans- ferred by being passed medially from tentacle to tentacle Feeding Differences in Metridium and finally dropped into the mouth. When adult Artemia were fed to large Metridium the response was restricted to the few tentacles involved in capture,, or to a fraction of a lobe of the oral disc. The opposing margins of a lobe pressed together enclosing the prey. The tentacles then relaxed and and the food was passed among adjacent tentacles toward the mouth. The prey was transferred to the larger central tentacles which deposited the food in the mouth. Often the Artemia was moved randomly and sometimes it was discarded from the edge of the disc. Small Metridium fed squid and Mytilus pieces, caught them with a group of tentacles. The entire group of tentacles then moved the food toward the mouth, the oral disc contracting until the food reached the lips. Food transfer did not seem to involve ciliary currents of the tentacles. When squid and Mytilus were fed to large individuals, the opposite sides of the disc lobe stimulated converged to enclose the food. The tentacles immediately surrounding the food began to wave, and a mucus coating with bubbles was produced around the food. Food and mucus were passes between tentacles into the cavity formed by the sides of the disc lobe where it was hidden from view. The food appeared again near the base of the lobe and Feeding Differences in Metridium from there slid rapidly into the mouth. The sliding may have involved both gravity and ciliary action. In general, the small Metridium do not respond to the smallest food, perhaps due to insufficient mechan- ical stimulation of the tentacles. They efficiently consume larger food, the extent of response increasing with the size and degree of activity of prey. Large anemones can easily capture numerous small food animals, Much more movement and time are needed to handle larger food. The great number and the arrangement of tentacles and the behavior of large Metridium are adaptive toward feeding on numerous microfood particles while the small anemones can manage pieces larger in relation to their body size. The deciding factor seems to be the distance between the mouth and tentacles and the corresponding difficulty in transferring particles to the mouth. These conclusions are in agreement with the observations made in previous studies. Natural Foods of Metridium In order to determine foods consumed by Metridium under natural conditions, animals were collected in the field, returned to the lab, and settled in clean aquaria provided with filtered running seawater. Batham and Pantin (1950) state that Metridium produce fecal pellets within 48 hours after feeding. In the present study, Feeding Differences in Metridium animals freshly brought from the field defecated in be- tween 24 and 48 hours. Pellets from 50 small, 15 inter- mediate sized, and 18 large Metridium were examined under a dissecting microscope. The pellets contained detritus and food animals in various stages of digestion. Re¬ cognizable remains of organisms were visually separated from the unidentifiable detritus. When necessary for identification, the remains were mounted in glycerine and examined under a compound microscope. Relative abundance of the different species or taxa was deter- mined from counts and from qualitative estimates of their percentage of the total biomass. To determine foods available, material was scraped from the pilings adjacent to the site where anemones were collected, and plankton tows were taken at the collecting site. These collections were examined, and relative abundance of organisms est- imated as above. Phytoplankton, unrecognizable detritus, and sand, abundant in the water, were not included as food sources. The small anemones each excreted several tiny pel- lets and a dark reddish-brown mucus. The total volume eliminated by each animal was approximately 0.1 ml, con- sisting of 65% mucus, 30% detritus and 5% animal matter. Large Metridium produced one or a few sizeable pellets, total volume for each animal averaging 1.4 ml. The Feeding Differences in Metridium percentage of detritus ranged from 40 to 60%. One day later the large anemones regurgitated large amounts of mucus containing bubbles which floated to the surface. Microscopic examination showed that mucus from both sizes contained high populations of bacteria. Occasional undigested phytoplankters and foraminifera occurred. The relative abundance of the organisms most fre- quently encountered in the environment and in the fecal pellets of small and large Metridium are compared in Fig. 2. The diet of small Metridium consists mainly of the three types of worms from the pilings. The re- mainder also consists largely of immediately adjacent benthos. In sharp contrast, the food of large Metridium represents mostly planktonic forms. However, the diet here showed a greater variety; additional food organisms not indicated as utilized by large Metridium in Fig. 2 due to their rare occurrance, include crab chelae, shrimp appendages, gastropod larvae, tunicate tadpole larvae, and barnacle nauplii. Fecal pellets from both large and small anemones included sponge fragments and spicules, crustacean fecal pellets (possibly from the guts of di¬ gested prey), and the exoskeletons of barnacle appendages. Most of the barnacle exoskeletons were probably molts captured by the anemone, however fresh mouth parts and a barnacle ovigerous lamella were also found on occasion. Feeding Differences in Metridium 10 A partially digested barnacle was seen being extruded by one small Metridium in the field. Also included in the pellets of large and small anemones were variable frac¬ tions of sand, barnacle and mussel shell pieces, sea urchin spines, anemone mesentarial filaments, and in the large individuals, several fish scales. One large anemone extruded intact chiton plates in the lab. Clearly there is a marked difference in the sources of the organisms eaten by small and large Metridium; benthic and pelagic diets, respectively, are indicated. Even most of the scattering of benthos including bryozoan and hydroid fragments taken by the large anemones could have been captured waterborne. Pieces of mussel and bar- nacle may have been broken from higher on the pilings and cascaded down. The food of the small Metridium is found on or in close proximity to the surface of the pilings. Certainly part of the dietary difference be- tween large and small anemones relates to the distance of the tentacles from the substrate in each case. In a typical feeding posture, the animals are bent upword from horizontal on a vertical face (Fig. 1d-e). Here the tentacles of small anemones are close to the pilings, less than 2 cm away and often brushing the surface. The tentacles of large Metridium are usually at least 15 cm distant from the pilings. The same general argu- ment holds for anemones on horizontal surfaces. Feeding Differences in Metridium 11 Previous investigators have assumed that large and small Metridium may select foods of different size. This may indeed be true to a degree, but present results indicate the two size classes also geed on the organisms most readily available for capture. Crab zoea larvae bulk large in the diet of large Metridium compared to their abundance in the plankton at the time of sampling. This may not reflect selectivity, but merely a greater number in the plankton when the anemone fed than at the time the plankton tow was taken. Summary 1. Studies were made of the feeding mechanisms and sources of food in natural populations of small and large Metridium senile on wharf pilings in Monterey, california during April and May, 1975. 2. Artemia larvae and adults, the copepod Tigriopus, and squid and Mytilus pieces produced different responses in the small and large anemones when dispensed to tentacles at various distances from the mouth. Small Metridium (3-15 cm in column height) captured food larger than 1 mm, the number of tentacles and por- tion of the oral disc involved increasing with increased size and activity of the prey. Feeding Differences in Metridium Large Metridium (20-35 cm in column height) cap- tured Artemia larvae and Tigriopus and held them in the tentacles, closing completely later. Larger prey re¬ quired more time and tentacular movement to be trans- ferred to the mouth. The large Metridium seem ideally adapted for cap- ture of numerous planktonic organisms and the small anemones seem better suited to larger prey in relation to their body size. 3. Microscopic examination of the facal pellets pro- duced in the lab revealed their contents to be mucus, bacteria, animal remains, and unrecognizable detritus. Small Metridium utilized mainly benthic organisms from the wharf pilings as food. Nematode, polychaete, and nemertean worms constituted the bulk of their diet. Food organisms of the large Metridium consisted largely of zooplankton. Copepods and crab zoea larvae were the most abundant organisms in the plankton and in the diet of the large anemones. These results show a marked difference in the food sources of the two sizes of Metridium. Feeding in each case appears to be on those organisms most readily avail- able for capture. The tentacles of small Metridium are in close proximity to the surface of the pilings and its characteristic benthos. The large anemones, with 12 Feeding Differences in Metridium tentacular crowns distant from the pilings, are well adapted to capture of planktonic organisms. Acknowledgements My warmest thanks go to Dr. D.P. Abbott and Chuck Baxter for numerous suggestions, identifications and a great deal of time patiently and cheerfully given. 13 Feeding Differences in Metridium Literature Cited Batham, E.J. and C.F.A. Pantin. 1950. Phases of Activity in the sea anemone Metridium senile and their re- lation to external stimuli. J. Exp. Biol. 27: 77-99. 1951. The organization of the muscular system of Metridium senile. Quart. J. Microscop. Sci. 92: 27-54. 1925. The feeding habits of the sea anem- Elmhirst, R. one, Actinoloba. Scot.Naturalist 12, No. 3: 149- 152. 1955. The sea anemones of central Cal¬ Cadet. Hand, Wasmann J. Biol. Part 3(13: 189-251, ifornia. 1955). The reactions of Metridium to food Parker, G.H. 1896. and other substances. Bull. Mus. Comp. zool. Harvard 29: 107-119. 1905. The reversal of the effective stroke of the labial cilia of sea-anemones by organic sub¬ stances. Am.J. Physiol. 14: 1-5. 1917. Actinian behavior. J. Exp. Zool. 221 193-229. Pax, F. 1928. Anthozoa. Die Tierwelt Deutschlands, Jena, Tiel 4: 189-240. Stephenson, T.A. 1935. British sea anemones. Ray Soc. London 429p. Feeding Differences in Metridium Figure Captions Figure 1 Diagrams of Metridium senile. a-c Oral views showing outline of disc and arrangement of tentacles (Circles indicate approximate positions of tentacles not shown). a) small individual (less than 3 cm in column height). b) intermediate size individual (10- 15 cm). c) Large individual (20-35 cm). d-e Typical feeding postures of small (d) and large (e) individuals. Stippling represents capitular tissue. Figure 2 Relative abundance of organisms in the environ- ment and in fecal pellets of small and large individuals of Metridium senile. p - present; quantity small, or difficult to measure. 00%0. 00 Fege Bod 20 06 60 580 0 900 o. Geee oo 0. 238228 10 - 10 - 5 . 3 0 uo1yup 90 1 5 c 5 oa sbuilid Figin 2 2 5 - poa - a 51 55. Jo Soutueg