A BEHAVTORAL INVESTIGATTON INTO THE FUNCTIONS OF THE FIRST AND SECOND ANTENNAE OF PAGURUS GRANOSIMANUS (STIMPSON, 1859), AND PAGURUS SAMUELIS (STIMPSON, 1859 ). SALLY K. ROLLEFSON Hopkins Marine Station of Stanford University Pacifio Grove, California Running Head: Functions of Pagurus Antennae Introduction The very prominent and active first and second antennae of the various species of Pagurus, make them an obvious subject for a functional behavioral study. The lack of literature and the many contradictions in what literature there is on the subject, also encourages such a study. This paper will attempt to explore some of the possibilities of such a problem; particularly with respect to chemoreception, rheotaxis, and taotile reception. Studies were made at Hopkins Marine Station, Pacifio Grove, California, during April and May of 1965. Two species of Pagurus, P. Samuelis and P. granosimanus, were used. Both species are common in the intertidal regions around China Point, Monterey Bay, with P. granosimanus found slightly lower insthe intertidal where it is subjected to stronger currents than P. samuelis. Methods The primary method employed was direct observation of intact crabs, both in the field and in the laboratory, and later of crabs subjected to various extirpations of antennae and other appendages. Operations were performed on unanesthetized crabs which had been poked out of their shells with a probe, after a hole had been hammered in the side of the shell. Crabs were allowed to return to their shells immediately after the operations, which were performed with a pair of fine scissors. Antennules were removed down to the first articulation next to the statocyst, and in one group the statocysts were also removed. Removal of the statocysts did not appear to hamper a crab particularly, but such crabs were very slow to return to their shells. In attempts to demonstrate chemoreception, a technique sim- ilar to that described by Barber (1961 ) was used. Blotters were impregnated with the various test solutions and added to a basin containing the crabs to be tested. Food, such as fresh- crushed Mytilus edulis or Tegula funebralis, was also added to tidepools and tanks, and observations made. To examine left-right tendencies å "Y" of rocks was set up in a tank and orabs were individually tested for possible directional preferences. To avoid light effects, these tests were performed indoors and the tank was turned around after each trial and the orab tested again. Each crab was tested at least twice. For the rheotactio studies, a procedure similar to that used by Luther (1930) was followed. A glass dish approximately 15 centimeters in diameter was anchored in the center of a larger glass dish approximately 30 centimeters in diameter, to provide a convenient channel in which currents could be easily produced and reversed with a direoted stream of water. Each crab was tested four to five times in succession by means of reversing the eurrent when a response was elicited. When the crab did not respond to the current change after two revolutions about the dish, the current was reversed and the lack of response recorded. The normal response of both P. granosimanus and P. samuelis to a current is to move into it, at first facing the current, then, as the strength of the current increases, turning sideways, and finally, backing into the current in short hops. When the crab can no longer move against the current, he relaxes and is carried to a point where he can again move against it. In this paper, such behavior is considered a positive rheotactic response. Results Crabs lacking antennae appear able to lead close to normal lives, and recover quickly from the operation. Mortality was not noticeably heightened in the operated crabs and most survived at least five weeks in the laboratory. Operated crabs did show an increase in aggressive behavior toward other crabs, however, and were slow in returning to their shells after operation. There was no appreciable differenoe in the dispersal times of operated and intact crabs, but operated crabs exhibited less exploratory behavior than intact crabs. Crabs without second antennae were especially non-exploratory. It was generålly observed that the second antennae are closely to visual activity, being stimulated by sight into an exploratory and tactile response. The antennae are quickly withdrawn on contaot with objects, and are seen to follow objects passed in front of the orab, even from the other side of a glass partition. Blinded crabs dod not exhibit such a following response, although they do avoid objects and other crabs. Grabs which were blinded and without either pair of antennae also avoided objects in their paths without any apparent bodily contact, however, which would indicate a complex sensory system involving more than just the eyes and antennae. The second antennae have also been observed snagging escaping food partioles. Crabs without second antennae showed a slight tendency to extend their walking legs wider and further in front of them than unoperated crabs. No evidence for chemotaxis in the two species of Pagurus was found. Even intaot orabs do not exhibit a directed response, either to food, in the form of mussel or snail meat, or to such chemicals as acetic acid, beta alanine, quinine, dextrose, maltose, sucrose, or lactose. In no case was andescape response to a predator found. Blinded crabs showed no response to food dropped directly in front of them. There was no apparent left or right preference, either in intact crabs of crabs without one or both first and second antennae. The number of crabs tested ranged from 25 to 35 per group, with a total of 442 orabs tested in all. Figure 1 shows the results of the rheotactic tests. Positive rheotactio response is virtually eliminated in P. granosimanus lacking either all of the first antennae, or the brushes of the first antennae. Extirpation of only the right or left first antenna also showeddå lowered number of positive responses; out of twenty trials five crabs gave a positive response only 60 per cent of the time without the right antennule, and without the left antennule the same number gave 65 per cent positive responses. Extirpation of the second antennae or of the maxillipeds had no effect. P. samuelis, on the other hand, shows no such loss of rheotaxis with removal of the first antennae. The P. samuelis o n210 T=50 2 2 80 60 867 40 20 INTACI P SAMUELIS n230 1o0 TEIIO 80 94% 2 60 40 20 TACT n210 T250 84 NO ANT. n210 T-90 86 NO ANT, NO ANT n220 T200 92 n230 TE 146 27 NO ANT n=20 T2100 35 BRUSHES ANT n210 1=50 96 NO MXPD n220 T2100 87 NO MXPD FIG. I crabs were much slower to respond to the current when a response was observed, however, than were the P. granosimanus. A total of 110 crabs were tested, and within each group there was a remarkable consistency of response. It was also noted that both species of Pagurus continually align their antennules into currents, even when backing into them. Brock (1930 ) found similar behavior in Carcinus maenas. Discussion In studies on chemoreception in other decapods, Hodgson ( 1958), and Case and Gwilliam ( 1961 ) found little evidence for chemoreception by either pair of antennae, but did find very sensitive receptors on the dactyls of the walking legs. In the present study, no evidence for a directing chemoreceptive organ was found, but in tests with glaeial acetic acid it was noted that the dactyls and mouth parts were apparently more sensitive than other parts of the crab. The fine brush structure of the antennules and their orientation with currents would make them a logical chemoreceptive organ, but no indication of such a function was found in this study. The most interesting results were those on rheotaxis. It was a surprise to find that two animals of the same genus, living at times in the same tidepool, should display such a difference in rheotactic behavior. Luther (1963 ) found similar differences between certain decapod genera, though. P. granosimanus is found in areas of stronger current than is P. samuelis,which may help to explain the difference. P. granosimanus, because of its greater exposure, may have lost all current sensitivity except in the antennules. P. samuelis may have body hairs which are current sensitive, or it may simply react to pressures on its shell. A neurophysiological investigation is the next logical step. Summary lons of the first and second A behavioral study of th antennae of Pagurus granosimanus and Pagurus samuelis was made. The primary method was extirpation and comparison with intact crabs. No chemotazis or lefteright preference was observed. Rheotaxis was found to occur in both species, but antennular involvement was only evident in Pagurus samuelis. Literature Cited Barber, S., (in Waterman, T. Ed. (1961 Physiology of Crustacea II: 112-113 Academic Press, N. Y. (681 p%2 ) (1930 Brock, F. Das Verhalten Der Ersten Antennen von Brachyuren und Anomuran in Bezug auf das Umgebende Medium Verg. Phys. 11: 774-790 Case, J., and G. F. Gwilliam (1961 Amino Acid Sensitivity of the Dactyl Chemoreceptors of Carcinides maenas 121: 449-455 BIOI. Bull. 1958 Hodgson, Edward Electrophysiological Studies of Arthropod Chemoreception Biol. Bull. 115: 114-125 1930 Luther, W. Versuche über die Chemorezeption der Brachyuren 12: 177-206 Verg. Phys. Luther, W. „( 1963 ) Versuche über die Funktion der 1. Antenne von dekapoden Krebsen als Strömungssinnesorgan Helgolander Wiss. Meeresuntersuchungen 844): 321-332 Legend for Fig. 1. Percentage of positive rheotactic responses observed in A) granosimanus. (n- total number P. samuelis, and B) P. crabs tested per group; Te total number of trials per grou