ADDITIONAL INFORMATION, IF ANY, CONCERNING AUTHORS, ADDRESS, TITLE, OR CITATION DATA PLEASE TYPE ABSTRACT DOUBLE SPACED BELOW JESSEE, WILLIAM F. (Hopkins Marine Sta. of Stanford Univ., Pacific Grove, Calif., USA). Studies of homing behavior in the limpet Aomaea scabra (Gould, 1846). The Veliger Homing behavior of A. scabra (Mollusca: Gastropoda: Prosobranchia) from rocky shores was studied in field and laboratory. Populations residing 7.1 and 2.4 feet above mean lower low water showed no sig- nificant differences in höming ability. All size classes above 5-6 mm. in shell length showed equal homing abilities; smaller ani- mals often did not exhibit homing. Scrubbing the substratum with a fiber brush, or treatment with 32% NaoH had no significant effect on homing tendency. Chiseling of the rock surface caused a drop in inci- dence of homing. This suggests that homing behavior is mediated throug perception of the topography of the substratum, and is not a chemo- tactic response. Removal of the cephalic tentacles reduced the ten- dency to home, while removal of the eyes did not.--Author. PLEASE DO NOT TVPE BELON THAS LINE 2 C Studies of Homing Behavior in the Limpet Acmaea scabra (Gould, 1846) by William F. Jessee Hopkins Marine Station of Stanford University, Pacific Grove, Calif. 93950 0 ove a Tho tendeney of membors of the species Aemasa abra (Gould, 1845) to re repeatedly to a spocifie location on their rock zubstratum, i.c. homing, has been reported from time to timo, iu-tho-litemo seh Ouof tho first stud on the Pacific species of Acmaea, by Wells (1917), gives the impression that homing is a highly individual trait, even among mombers orithe single species A. scabra. However, the tendency to home was apparontly not studied in moro than a few esentatives of each spocios, and tho frequency of 20 ag behavior was not ovaluated. To attem ts at such an evaluation were subsequently reported (Howatt, 1940; Villee and Groody, 1940). Hewatt's study of thirty-ono A. scabra on granite rocks reveoled homing behavior in all thirty-one individ- uals. On the other hand, Villeesand Groody comcluded from their study of 86 members of this species on a sandstomo substratum, that there was no ovidence for homing behavior. This latter study, however, was roe stricted to observations made principally as low tide, a time when there is little or no activity in tho lim- pot population. All animals that had not beem observed away from their homosite during the period of study ro ignored. In a more recent quantitative study by Brant (1950), A seabre 298 marked wero observed over a period of 34 days. Homing behavior was reported in 58.79 of this test population. Proliminary observations by tho am 726 O C * Sil ese 0 . author collirmod thoso findings. The principal object of this study was to inves- tigato tho mochaniam involved in homing bchavicr. In addition two othor aspocts of heming which have rocoived little investigation aro included in this study. Toso aro tho homing behavior of animals from high and low elevations in tho intortidal, and the bohavior of different size classes of limpots. t m METHODS AND RESOLTS Who obsorvations and oxperiments doscribed below wero made at Pescadoro Point on tho Monteroy Peninsula, California during Apzil and May, 1966. Tho first series of experiments performed was dosigned to detoct any difforences in the homing abile ity of high and low populations, and in that of large and amall animals. Ono hundred individuals of intormodiato size, betwoen 10 and 15 mm. in length, wero measurod, und tho animal and its homesite marked with red nail polich. Fifty of those amimals woro on a homizental granito surface with a median olevation of 62.4 ft.. e — this population will be roferred to as the Wiow" group. Tho other: fey amimals, hereafter called the "high" population, were on a gently sloping rock with a modian olovation of 97.1 ft. After waiting throo days to insure that the position marked was indoed the animal's homo, a procoduro that was followed in all subsequent oxperiments, 30 in each group wero dise -3 William F. Jessee placed randomly 3 - 4 cm. from their homesites. The other twenty in each area served as controls. The animals were observed after 24 hours, and as shown in Table 1, no signi¬ ficant difference was seen in the homing ability of the high and low groups. A similar experiment was then carried out in an area of intermediate elevation, +3.9 ft., to compare the homing behavior of large animals, greater than 15 mm. in length, with that of small ones between 6 and 10 mm. long. Here, too, no significant differences were observed, as can be seen from Table 2. However, a less complete study suggested that animals smaller than 6 mm. usually do not home. Experiments in the laboratory, utilizing glass plates over which limpets had been allowed to move, revealed no residual mucus in the form of trails as determined by the India ink test of Peters (1964). Nonetheless, field exper¬ iments were performed to investigate the possibility that mucus or some other chemical substance on the surface of the rock was the agent active in governing homing behavior. In the first type of experiment, a stiff fiber brush was used to scrub a 14" strip around the homesites of 20 previously marked animals. The areas surrounding 15 other animals, which served as controls, were left unscrubbed. All animals were then displaced 4-5 cm. from their homes -4- William F. Jessee to a position outside this strip. Their locations after 24 hours were noted, and are presented in Table 3. A variation of this experiment was also carried out in which the home itself was included in the area scrubbed. This was to eliminate any possible attractant on the home, as well as "trails" leading to the home. As shown in Table 3, neither of these treatments completely destroyed the ability to home. A second group of experiments was performed to remove chemical substances. All animals in a one foot square area were removed and the rock painted with a solution of 32% NaoH. The surface was then rinsed repeatedly with copious quantities of seawater until the pH of the wash water was near neutrality. The animals were then replaced within 4 cm. of their homes. Five animals outside the treated area were also displaced a similar distance and utilized as controls. At the end of the customary 24 hour waiting period, 8 of the 10 animals in the experimental group had returned to their homes. Having established that the removal of chemical sub¬ stances on the rock had no noticeable effect on the homing ability, experiments were begun in which the topography of the rock surface surrounding the home was altered. A geologist's pick and a chisel were used to create a strip -5 William F. Jessee 1" wide around the homes of 25 animals. The topography of the rock in this strip was totally changed. The animals were then replaced 4 cm. from their homes, and outside the chiseled area. Fifteen other animals were similarly dis¬ placed, but the topography of their surroundings was left unaltered. This latter group served as a control. Within 24 hours only four of the 25 experimental animals had homed. Seventeen of the 25 were not in their homes, the remaining four having disappeared. Some additional evidence suggesting the involvement of topography in the homing mechanism was an experiment performed with a small group of animals at China Point. Seven animals were marked and displaced the customary 4-5 cm. from their homesites. A hammer and chisel were then used to create a strip about 1" wide between the animal and its home. As the tide came in, the animals were ob¬ served to move to the edge of the chiseled area, then travel along the edge of this area until they reached its end. At this point they resumed their travel toward the homesite, six of the seven returning by the next low tide. This reaction to territory made unfamiliar by alteration suggests that recognition of some element of topography may be involved in the homing mechanism. C -6- William F. Jessee DISCUSSION If populations at the high and low levels are studied using animals of comparable size, there is no significant difference in the homing ability of the two groups. Nor is there any significant difference in the homing ability of animals of different sizes at the same intertidal level. These observations confirm earlier ones by Haven (1966). However, it was also observed that animals of less than 5-6 mm. in length usually do not home. Such animals are found mainly in the lower intertidal. The inclusion of these extremely small animals in earlier studies of homing behavior may account for some of the discrepancies in the findings of this study as compared to these earlier works. The results of the experiments designed to elucidate the mechanism of homing suggest that it is related to the perception of the topography of the rock surface in the vicinity of the limpet's homesite. The fact that the ani¬ mals would not cross an area of unfamiliar topography, but went directly home upon reaching familiar territory points to a "memory" of the convolutions of the rock surface. It is, however, possible that some other factor in the environ¬ ment which was not investigated is responsible, at least in part, for homing. Light or its polarization pattern, for 16 -7- William F. Jessee instance, could be such a factor. The probability that the mechanism is indeed involved with topography is in good agreement with the tentative results reported by Galbraith (1965) on the mechanism of homing in Acmaea digitalis Eschscholtz 1833 and Lottia gigantea Sowerby 1843. The high percentage of animals which homed after treatment of the rock with sodium hydroxide argues against the possible use of some chemical agent as a homing quide, as the concentrated alkalai would be expected to dissolve to or denature any substances deposited by the limpets, While other external factors, such as polarized light or water currents, have not been ruled out, it is doubted that the animal possesses the sensory equipment to utilize such highly variable sources for orientation. Work should, however, be carried out to examine these possibilities. The implications of a homing mechanism based on a knowledge of topography, in an animal such as the limpet, demands additional investigation. 8 ede O C WWia k. Zessea 18 ADDENDUMy were Two additional series of experiments haveesatl ater completed in an effort to determine what portion of the sensory apparatus is involved in homing behavior. The first experiments, designed to determine the role of the cephalic tentacles in homing, ereperformed at Pebble Beach, San Mateo County, California in January and February, 1967. In these experiments sixty animals were marked in the customary manner and left for three days to insure that the position marked was the home spot. Then a small typewritten number was affixed to the shell of each animal, and the same number placed adjacent to its home using Duco cement. The animals were then brought into the laboratory, where all were anesthetized in a solution of magnesium chloride isotonic with seawater. Both cephalic tentacles were then excised from forty animals, distal to the eyespots. The remaining animals were uninjured and were utilized as controls. All animals were then placed in a salt water aquarium for a period of four to five days to allow recuperation of the operated animals. The animals were then returned to the beach during a period of low tide, and placod within three to four centimeters of their own homesites. The locations of the animals at the end of 24 hours, or two high tides, were observed and are summarized in table 4. Of those animals which could be located at the end of the 22 hour period, 293 of the experi- mental group had homed, as compared to 75% of the controls. O Dilliau k Besree 1 The second series of experiments was performed at Moss Beach, San Mateo County, during April and May, 1967. and was designed to determine whether or not the eyespots are utilized in homing. Sixty animals were marked and in- dividually numbered as before. After the usual waiting period the limpets were brought into the laboratory, where all were anesthetized with the magnesium chloride solution. A dissecting needle which had been heated to redness was then employed to cauterize both eyespots on forty of the animals, the remainder being utilized as controls. After a four to sixday period of recovery in the aquarium all the animals were returned to the field and replaced within three to four centimeters of their homes as before. The positions of these animals were observed at the end of 24 hours, and are also summarized in table h. The eyespot is apparently not utilized in homing. Exam- ination of the four experimental animals which failed to home indicated that in these, substantial damage had been done to the cephalic tentacles. In those animals in which no damage was observed, there was 100% homing. The loss of the cephalic tontacles appears to have a statistically significant effect on homing. This observation is, however, open to question. It is a distinct possibility that the decrease in the instance of homing was a result of trauma or other factors. The fact that some 29% of the experimental group returned to their homes despite the loss of the tentacles seems to substantiate this possibility; the presence of alternative systems which may be utilized WWlian K. Sesgea in homing is another possible explanation. hsephalie tenteeles ere in feet invelree in heningthieweuld anpete ferther imple that homing pehavior-isbasee-enthe mereepttorofthetopogrephyofthesubstrate. O O C Willau k. Jessee 16 SUMMARY 1. Homing behavior of Acmaea scabra was studied in the field with respect to the intertidal height of the popula- tion, the size of the individuals, and the mechanism involved in homing movements. 2. No significant difference was found in the homing ability of populations in the high intertidal region as compared to those at lower levels. 3. The size of the animals was found to be unimportant in determining their ability to home, except that extremely small animals, less than 5 to 6 mm. in length, were usually found to be non-homing. 4. The possibility that chemical substances on the rock surface are the agents of homing behavior appears highly unlikely. 5. Although a few other environmental factors were not coerimentally eliminated, the evidence presented indicates that the topography of the rock surface is utilized in homing by A scabr: 6. (From addendum) Excision of the cephalic tentacles anificantly reduced the tendency to home, while destruction si of both eyespots had practically no effect on the incidence of homing. O C 0 Dillian f Jegse 17 1 ACKNOWLEDCEMENTS The author wishes to express his thanks to Drs. J. H. Phillips and D. P. Abbott, and to Mr. Stoner L. Haven for their advice and assistance; the advice and keen interest of the late Prof. V. C. Twitty was also deeply appreciated, Thanks are extended to the Monterey Foundation and Del Monte Properties, Inc. for permission to work in the Pescadero Point area, This work was made possible by Grant GY-806 from the Undergraduate Research Participation Program of the National Science Foundation, and by a grant from the Arizona Academy of Science. C Bieagges 12 18 --mr LITIERAIURE CITED Brant, Daniel - 1950 - "A Quantitative Study of the Homing Behavior of the Limpet Acmaea scabr. published work in the Department of Zoology, University of California, Berkeley, California. Galbraith, Robert T. - 1965 - "Homing Behavior in the Limpets Acmaea digitalis and Lo gigantea" Amer. Midl. Nat. 74 (1): 245 - 246. Haven, Stoner L. - 1966 - Personal communication. Hewatt, Willis G. - 1940 - "Observations on the Homing Limpet, Acmaea scabra Gould" Amer. Midl. Nat. 24 (1): 205 - 208. Peters, Ronald S. - 1964 - "Function of the Cephalic Tentacles in Littorina planaxis Phillipi (Gastropoda: Prosobranchiata)" The Veliger.7. (2): 143 - 148. Villee, C.A. and T.C. Groody - 1940 - "The Behavior of Limpets with Reference to their Homing Instinct" Amer. Midl. Nat. 24 (1): 190 - 204. Wells, Morris M. - 1917 - "The Behavior of Limpets with Particular Reference to the Homing Instinct" Journ. Anim. Behav. 7, (6): 387 - 395. O Williaen f. Tessen 1 Table 1. Homing behavior of A scabra populations at high and low intertidal locations. Observations made 24 hours after displacement of experimental animals. Table 2. Homing behavior of large, greater than 15 mm. in length, and small, 6 - 10 mm. in length, individuals of A. scabra. Observations made 24 hours after displacement of experimental animals. Table 3. Homing behavior of A. scabra following scrubbing of home and/or surroundings. Observations made 21 hours after displacement of experimental and control groups. scabra following excision Table 2. Homing behavior of A. of both cephalic tentacles or bilateral de- struction of the eyespots. Observations made 24 hours after replacement of experimental and control groups. C Low Animals Displaced — Low Control Animals igh Animals Displaced — High Control Animals — 5a1 30 20 30 able 25 19 27 20 Not SlVan ee Missing 2 O O (6 Tor e 30 Animals Displacod — Largo Control Animals Small Animals Displaced — - Small Control Animals Toi 20 15 40 50 able 2 Hom 19 15 36 No Hom SM Mis: 0 O Sürroundings scrubbed No sezabbing — Home and surroundin serubbed No serubbing al 20 10 30 20 able rr.. n 10 — 10 23 16 Not! O See Miss O ka - Cephalic Tentacles Eised — Controls — Eyospots Destroyed Controls — Total 10 20 10 20 able Home 9 12 28 11 — Liliau 6. Yesses 10. Home Mis ing 22 —