MACROSCOPIC ALGAL FOODS OF LITTORINA PLANAXIS AND L. SCUTULATA by Arthur Lyon Dahl Hopkins Marine Station Biology 175h Dr. Blinks May 30, 1964 MACROSCOPIC ALGAL FOODS OF LITTORINA PLANAXIS AND L. SCUTULATA The upper littoral periwinkles of the central California coast are often found in association with a number of the higher intertidal macroscopic algae. This paper examines the place of these algae as a food source for Littorina planaxis Philippi, one of the highest intertidal animals, and for its somewhat lower though overlapping relative L. scutulata Gould. The studies and observations upon which this paper is based were made on the Monterey Peninsula, on the central California coast, especially at Mussel Point in the vicinity of the Hopkins Marine Station, and near Pescadero Point. The coastline in these areas consists principally of granite boulders and outcroppings interspersed with sandy beaches, and heavily overgrown with larger algae through most of the littoral zone. Observations were made during May, 1964, and therefore do not reflect any seasonal fluctuations that might take place. Despite the fact that both Littorina planaxis and L. scutulata are a conspicuous part of the upper littoral fauna over much of the Pacific coast of North America, (2) almost nothing has been published about them up to now, and while the eastern U.S. and European littorines are generally better known, information on food and feeding patterns is very scarce. North (1954) analyzed size distribution, erosive activities, and gross metabolic efficiency of both L. planaxis and L. scutulata. Castenholz (1961) used L. scutulata as well as Acmaea spp. in his studies of grazing effects on diatom populations. From his observations, he assumed "that the primary food of these gastropods is diatom material," with blue green algae the principal food in the "supra-littoral" fringe. Ricketts and Calvin (1952) state that L. planaxis feeds on detritus and microscopic plants scraped from almost bare rock, and that "certain of the rockweeds (Pelvetia or Fucus) serve the young periwinkles as a sort of nursery, for it is on their fronds and stems that the young will nearly always be found, "(p. 20), an observation for which no evidence has been seen during this study. Concerning other species, Newell (1958) noted that Littorina littorea ate surface deposits of diatoms and small algae, and also browsed on Ulva and Enteromorpha. The answers to three questions were sought in this study. Can these snails eat macroscopic algae? Which algae, if any, are normally part of their diet? How important is macroscopic algae in their diets? A number (3) of approaches were used in the attempt to answer these questions. Numerous field observations were made to determine what algae the snails had access to, and if they were ever in contact. Principal proof of feeding was based on examinations of the stomach contents of disected snails, from which ingested tissue fragments could be compaired with scrapings from the algae on which the snail was found, or on which it was believed to have fed. In a few cases, feces were also examined for recognizable algal fragments. Several types of laboratory experiments were attempted to supplement field observations. In a few instances it was sufficient to place a winkle together with a dampened piece of algae in a dish. Better results were obtained in an aguarium the contents of which were kept wet but not submerged by a fine sea water spray. An aquarium was also rigged to duplicate tidal fluctuations, but in no case were conditions similarenough to those in the field to get normal responses for any length of time. North (1954, p. 193) noted that food cycled completely through the digestive tract in from 24 to 6 hours. Thus the stomach contents of any snail kept for six hours with a single type of algae, or starved for six hours and then placed with an alga, could come only from that alga. This provided a simple and reliable check for other types of observations. A number of difficulties were encountered in the 0 (4) attempt to induce feeding in the laboratory. L. planaxis has a strong tendency to crawl to the highest part of the dish or aquarium, especially if placed in water. Individuals of both species would frequently cease activity if left undisturbed for more than 6 to 12 hours. Natural conditions, especially with respect to tidal and diurnal fluctuations, proved almost imposible to duplicate, yet it would have been even more difficult to keep track of individual snails in the field, or to eliminate unwanted food sources there, especially the ubiquitous GATGOR (Green Algae That Grows On Rocks, a term applied to the microscopic flora. Work on this aspect of littorine feeding has been done by Mr. Michael Foster). Experimental animals had to be run singly in experiments where starvation was a factor because of the N tendency of snails to feed on each others' backs. Additional complications resulted from epiphytic algae growing on the test specimins, and from the rapid decay of certain types of algae, especially Iridophycus flaccidum and Laminaria Andersonii, under conditions available in the laboratory. A number of criteria were set up by which feeding on any particular alga could be substantiated with a reasonable degree of certainty. At least two of these three standards had to be met before a snail could be considered to have "fed" on the alga in question. These O O (5) were as follows: (a) the snail was observed on the alga, preferably actively moving, and with the radula active; (b) whole pieces or fragments of tissue were found in the stomach or feces exactly matching in cell size, shape, structure, color, type of chloroplasts, etc., scrapings or known forms of the alga in question (usually determined by direct comparison); (c) the animal was starved (kept with no signs of feeding or no food) for at least six hours prior to feeding or disection, and had stomach contents not dissimilar to fragments of the test alga. Most of the observations of feeding on any particular alga are based on only a small number of individuals because of the time involved in making a positive determination of feeding, and, in some cases, because such feeding itself may occur only rarely. Some algae are found over only a small part of the range of L. planaxis or L. scutulata, or snails may feed on them only under particularly favorable conditions. It was assumed, however, that positive proof of feeding on an alga, even if based on only one individual, would indicate a species capability if not a preference, and that once such a capability was demonstrated, snails observed on such an alga were probably feeding on it. Littorina planaxis is generally found in the upper regions of the intertidal zone, on "bare" rocks normally 66 O (6) moistened only by splash. The height of this region varies considerably with the amount of exposure to surf. Littorina scutulata occurs somewhat lower, and in more protected areas, ranging from about +2 feet up to about +6 feet in some areas. It is frequently found in moderate to heavy growths of algae, and smaller individuals occur in large numbers in some horizontal Balanus beds. The ranges of the two species of littorines overlap considerably in some areas. Both species are known to depend on microscopic algae (diatoms, greens, and some blue greens) as a principal food source (Castenholz, 1961; Foster, unpublished), especially where macroscopic algae is scarce or absent, as it is over much of the range of L. planaxis. However, there are areas in which considerable macroscopic algae is available, and it is with these areas that this study is primarily concerned. A number of common algae occur in the vicinity of littorine populations, especially Cladophora trichotoma (C.A. Agardh) Kützing, Endocladia muricata (Postels and Ruprecht) J.G. Agardh, Fucus furcatus C.A. Agardh, Gigartina Agardhii Setchell and Gardner, Gigartina cristata (Setchell) Setchell and Gardner, Iridophycus flaccidum Setchell and Gardner, Pelvetia fastigiata (J.G. Agardh) DeToni, Porphyra perforata J.G. Agardh, Rhodoglossum affine (Harvey) Cladophora is generally found Kylin, and Ulva spp. 1. Nomenclature of algae follows G.M. Smith, Marine Algae of the Monterey Peninsula California. a? (7) in cracks, crevasses, and tidepools, on horizontal surfaces, sometimes together with Ulva; Endocladia, Rhodoglossum, and the gigartinas are frequently associated together in crevasses and on rock surfaces, as well as in scattered individual clusters; Pelvetia, Fucus, and lridophycus are commonest on sloping surfaces at slightly lower levels; Porphyra often covers the tops of boulders and outcroppings. One area, in which many of the field observations included in this study were made, includes all of the above species except Fucus, and in addition has large populations of both L. planaxis and L. scutulata. It is situated on a granite shelf west of Pescadero Point. at a height of about +4 feet, and was divided for survey purposes into two sections, each of approximately 2 square meters in area, The first is largely horizontal, with a number of crevasses up to 10 cm deep, and a small tidepool. Cladophora is the dominant alga, with some Endocladia and Rhodoglossum also present: the algae aree largely confined to the crevasses and around the tidepool, leaving the elevated rock areas bare. The second consists of the verticle west face of the shelf and some of the contiguous horizontal top, about 2 meters from the first area, and at the same level. It is almost entirely covered with algae, with Endocladia, Rhodoglossum, Pelvetia, and Gigartina cristata the most common species, (8) and the others present in lesser quantities. One survey showed that the first area contained 147 L. planaxis and 329 L. scutulata. Surveys were generally made in the morning while the snails were still active from their previous nights wetting, yet had had enough time to feed adequately on the algae. The feeding behavior of Littorina planaxis and L. scutulata seems to be a combination of random activity with preferential movement. In the laboratory, snails are frequently observed scraping their radula along a clean glass surface, and have been known to consume considerable quantities of paraffin and powdered carbon. In the field, the stomach contents of snails have revealed large qunatities of sand and rock particles, detritus, any number of algal types, and occasional small animals, often still alive and active. There seems, therefore, to be little or no selectivity in what is ingested from any given surface. However, field studies to be discussed later suggest that each species has, at least under some conditions, definite preferences for certain types of substrates or algae. The radula of littorines seems capable of tearing off quite large pieces of substrate material, and particles of considerable size have been found in the stomachs. Some of the smaller algae such as the filamentous Cladophora and the small monostromatous Prasiola meridionalis (9) are frequently found almost entire on the stomach of L. planaxis. The structure of the radula and other mouth parts seems to be similar for both specias, and, superficially at least, matched that of Littorina littorea as described by Fretter and Graham (1962). What the littorines digest out of what they take in is still uncertain. Undigested cells and tissue fragments are frequently found in the feces, supporting North (1954) in his conclusion that their digestive efficiency is low, with organic matter assimilation about 7 percent of consumption. Cell wall material seems especially conspicuous, at least in the stomach, and was often useful in identifying the algae on which the snail had fed. Littorina planaxis does not frequently come into contact with macroscopic algae; most of its range is well above the limits of algal growth. Thereeare areas, however, where it is found with all of the algae listed previously. In such areas, it shows a decided preference for a rock substrate (For the purposes of this study the term "rock substrate" includes both microscopic and encrusting algal forms; "algal substrate" refers to macroscopic algae with other than an encrusting thallus. The substrate is the surface to which the snail is attached, and on which, presumably, it is feeding). One survey of the area near Pescadero Point (10) showed only 2% of the L. planaxis on algal substrates; for another check under more favorable conditions (overcast sky), about 10% were observed on Cladophora. One individual collected and disected had a considerable quantity of Cladophora in its stomach and a filament hanging out of its mouth, so feeding was almost certainly taking place in these cases. L. planaxis has also been collected in a few instances on the thallus of what is either a high form of Rhodoglossum affine (the name used in this paper), or a juvenile form of Gigartina cristata. The stomachs of these snails contained a few fragments of tissue closely resembling scrapings from the alga, but in such small quantities as to be an insignificant part of its diet. Snails collected from patches of Prasiola meridionalis Setchell and Gardner on rocks covered with bird guano at Mussel Point proved to have large quantities of this alga, often whole thalli, in their stomachs and feces, in addition to quantities of a microscopic chlamydomonad. These are the only instances in the present study in which L. planaxis has been found to eat macroscopic algae under natural conditions. A number of laboratory experiments suggest reasons for this behavior. In one experiment, thalli of Endocladia, Rhodoglossum, and Porphyra were finely chopped with a razor blade and spread on the wet bottoms of petri dishes; dishes with whole thalli, and a dish with a growth of GATGOR (11) served as controls. A single freshly-collected L. planaxis was placed in each dish for 5 hours and then disected. The snails consumed large quantities of the chopped pieces whole, and also the GATGOR, while the stomachs of the control winkles were empty, even whentthey were attached to the algae. In an algal preference test run in an aquarium kept damp by spray, the few L. planaxis that did not crawl to the top of the tank fed either on rock samples or on soft, decaying pieces of Iridophycus flaccidum. There a There are several possible reasons why L. planaxis does not normally eat much macroscopic algae: (a) it is unable to "bite off" pieces of any but the softest of most finely divided algae; (b) it for some reason will not crawl on the "unstable" substrates provided by algae; (c) most macroscopic algae are somehow chemically disagreeable to it. Item (c) can be safely discarded, since L. planaxis showed no aversion to chopped algae. There is at present no evidence for or against reasons (a) and (b), although a brief examination of the mouth and radula showed no obvious differences from those of L. scutulata, which feeds much more on macroscopic algae. In contrast with L. planaxis, Littorina scutulata both encounters more macroscopic algae and eats more of it. For'instance)oinrthe survey near Pescadero Point cited above, 47% ofthe L. scutulata in both areas were (12) on algal substrates. There were marked preferences for certain species of algae. Of the 153 snails on algae, 72 were on Cladophora, 46 on Pelvetia, 20 on Rhodoglossum, and no more than 5 on any of the other species. Nor is this entirely a reflection of the relative abundance of the algae; Endocladia, the most common, had only 3 L. scutulata on it, and some of the others had fewer than their relative areas would suggest. Also, observations under more favorable conditions than those reflected in the above survey suggest that all these figures, especially for Pelvetia, are at times considerably higher. In other areas, L. scutulata is also found on Fucus and Porphyra. In addition, tidepool populations of L. scutulata have been observed scavenging on decaying algae such as Gigartina corymbifera and Prionitus lanceolata that had been left in the pools by very high tides. In these particular instances, the stomachs of snails collected from the algae were stuffed with partially decomposed algal material. Laboratory studies confirmed that L. scutulata will readily eat a variety of macroscopic algae. In addition to the Cladophora and Pelvetia mentioned above in which feeding was definitely observed, snails readily attacked Ulva spp., leaving only tattered shreads. In the preference tank previously mentioned, L. scutulata was observed on all algae in the tank (Pelvetia, Cladophora, (13) Gigartina cristata, Endocladia, Rhodoglossum, Porphyra, Ulva, Iridophycus, Laminaria Andersonii), and all individuals disected had macroscopic algal fragments ase in their stomachs (these were identified for Porphyra, Laminaria, and Ulva). Concerning the effects of littorine feeding on macroscopic algae, the situation seems to be similar to that noted by Castenholz (1961) for the effects of L. scutulata on diatom populations; the snails are able to prevent sparse populations from increasing, but do not have much effect on well established colonies. The completely bare areas between algal growths in which littorines are so frequently found (Castenholz, 1961) are probably kept bare by continual grazing, and the occasional presence of very young thalli in littorine stomachs supports this. In the study area near Pescadero Point, the Pelvetia is worn and indented, and in some places only the stubby remains of thalli can be found in the bottom of cracks, too tightly packed for the snails to reach. Even the Endocladia is undercut and trimmed back for about 1 cm above the rock surface in some areas bordering on bare rock. Whether these effects are due to the feeding activity of littorines is not possible to say without lengthy observations and measurements, but considering the quantities of cell material found in stomachs, the depredations observed intthe laboratory, (14) and the number of snails involved, the effects of feeding are by no means insignificant. However, well established algal growths seem to be able to hold their own successfully, at least for short periods. Two areas, one with Porphyra, the other with Cladophora and Ulva were caged with fiberglass netting and + inch mesh galvanized screen into two compartments, one with 25 L. scutulata, the other empty as a control (a third compartment with 25 L. planaxis was added for the Ulva, Cladophora test). After two weeks, no effects were observed that could be directly attributed to the snails, although measurements were not very accurate. The effects on different algae probably vary considerably, judging from the varying quantities of algal material in the stomachs of snails feeding on different plants. Porphyra and Rhodoglossum, for example, were generally found in much smaller quantities than the softer Pelvetia and decaying algae, or the more easily ingested Cladophora. This study was of course) limited in its scope because of the short duration of the observations. More statistical surveys and field observations on preferences are needed, covering a larger number of areas; the effects of littorine feeding on algae should be measured quantitatively. Much more has yet to be learned about the mechanisms behind preferences, and the differences in feeding habits of the two species when in the same (15) area under the same conditions. A number of possible sources of error need also to be considered and remedied if possible: in almost all cases the samples were small; study areas in the field were selected because of the prevalence of littorines feeding on algae; no attempts were made in the laboratory to take into account the previous conditioning of the snails in the field with regard to food; laboratory conditions were far from duplicating natural situations (in the most successful attempt, the spray-doused preference tank, the three algae on which the least feeding was observed were Pelvetia, Cladophora, and Porphyra). In summary, Littorina planaxis generally eats only microscopic foods and those macroscopic algae small enough to be consumed entire or nearly so. The reasons for this are not clear, but may be related to the structure of the mouth, or the preference for certain types of substrates. Littorina scutulata is frequently found eating macroscopic algae, especially Cladophora, Pelvetia, and other more easily ingested forms. The effects of this feeding are probably important in limiting the spread of new plants, and the effects on established thalli, while possibly considerable in some instances, appear to vary greatly with the species of algae. (16) WORKS CITED Castenholz, Richard W. 1961. The effect of grazing on marine littoral diatom populations. Ecology, 42: 783-794. Fretter, Vera and Alastair Graham, 1962. British Prosobranch Molluscs, Their Functional Anatomy and Ecology. London, The Ray Society, 1962, pp. 26-27. Newell, G. E., 1958. The behavior of Littorina littorea (L.) under natural conditions and its relation to position in the shore. J. Mar. Biol. Ass. U. K., 37: 229-239. North, Wheeler J., 1954. Size distribution, erosive activities, and gross metabolic efficiency of the marine intertidal snails, Littorina planaxis and L. scutulata. Biol. Bull., 106: 185-197. Ricketts, Edward F. and Jack Calvin, 1952. Between Pacific Tides. Third Edition, revised by Joel W. Hedgpeth. Stanford, Stanford University Press, 1952, pp. 12-20. Smith, Gilbert M., 1944. Marine Algae of the Monterey Peninsula California. Stanford, Stanford University Press, 1944.