Linsenmeyer Attachment of Chitons INTRODUCTION According to Arey and Crozier (1919), the mechanism by which Polyplacophora attach themselves to surfaces is suction, primarily from the mantle. Hoffman (1931) comments that the role of the ventral mantle and the foot are of equal importance in fastening to surfaces. Detailed research on this subject, however, is lacking. This investigation was designed to determine the forces required to remove several species of Polyplacophora from various surfaces, and to elucidate the mechanism by which they resist removal. MATERIALS AND METHODS Mopalia lignosa (Gould, 1846), Mopalia muscosa (Gould, 1846), Nuttalina californica (Reeve, 1847), Katharina tunicata (Wood, 1815), and Stenoplax heathiana (Berry, 1946) were collected intertidally off the Monterey Peninsula in California in April and May, 1974. After collection, undamaged chitons were held in running sea water aquaria until tested. Chitons were used within four days of collection and each animal was used in only one trial. Prior to testing, chitons were allowed to attach to the test surface for a period of two hours. The device sketched in Figure 1 was used to determine the resistance page 2 Linsenmeyer Attachment of Chitons to removal by a force parallel to the surface. Aproximately three grams were added to the container every two seconds until the animal broke free of the surface. The device had a mechanical advantage of two so that the force required to remove the chiton was taken as twice the weight of the container. To determine resistance to removal by a force perpendicular to the surface, a cloth-backed picture hanger was glued to the dried plates of an animal (using the contact glue "ZIP GRIP" 10, Devcon Co., Danvers, Mass.), and connected by a string to the device illustrated in Figure 2. Weights were added at the rate of three grams every two seconds. Following each test, the area of the foot and the total area of the ventral surface were determined by placing the chiton on a transparent grid. Resistance to removal is expressd as grams per square centimeter of foot area, since the foot proved to be the primary site of attachment. The relation between roughness of the surface to which the chiton was attached and resistance to removal was also determined. Tests were conducted using chitons attached to smooth glass or Plexiglas, #120 waterproof emery cloth having an average grit size of 120 u and flat natural granite. In order to evaluate the role of reduced atmospheric page 3 Linsenmeyer Attachment of Chitons pressure (suction) under the foot as a method of attachment, individual Mopalia muscosa, attached to Plexiglas plates, were placed in a vacuum desiccating jar. Piano wire, attached to the glued hook on the animal, exeited the jar through a small hole and was connected to the device illustrated in Figure 2. Tests were run at atmospheric pressure and under vacuums of 127 and 380 mm of mercury below atmospheric pressure. Additionally, chitons were allowed to settle on substrates with varying pore size: fritted glass with a pore diameter of less than 15 u, a 1 mm mesh wire screen, and Plexiglas containing numerous 3 m diameter holes. Their ability to cling, after two hours was tested. RESULTS AND DISCUSSION Figure 3 shows the resistance of chitons to removal by forces parallel to a Plexiglas surface, in g/cm of foot surface, for each species. Nuttallina, which inhabits rocks subject to heavy surf, exhibited a mean resistance to removal of 237 g/cm. This was more than double the resisting force of the three species occupying the most protected habitats: the two under-rock species, Stenoplax heathiana and Mopalia lignosa, had mean values of 64 and 94 g/cm respectively, and Mopalia muscosa, found on the top surface of protected roacks, had a mean value of 112 g/cm. Katharina tunicata, which, like page 4 Attachment of Chitons Linsenmeyer Nuttallina, occupies a surf-exposed habitat, had a comparatively high mean resistance to removal of 146 g/cm. Thus, the resisting force had a direct adaptive relationship to the habitat occupied by each of the species. It was observed in all species that the plates and mantle were forcefully displaced as the loading increased, while the foot remained fixed. This strongly implies that the foot plays the major role in attachment. The lateral forces required to remove Mopalia muscosa from the three types of surfaces are shown in Figure 4. Mopalia muscosa had a mean resistance of 112 g/cm when settled on Plexiglas, 144 g/cm when on #120 emery cloth and 223 g/cm when on granite. The animals had a mean resistance to a vertical (lifting) force of 82 g/cm on natural rock. Thus substrate roughness had a significant effect on the resistance to detachment by forces both lateral to and normal to the attachment surface. During the tests using forces normal to the surface, the plates and mantle lifted off the surface by as much as 5 mm with¬ out detachment of the foot, again implicating the foot as the organ of attachment. No significant differences were detected in the resistance of Mopalia muscosa to lifting forces applied at atmospheric pressure and under veuums of 127 and 380 mm of mercury below atmospheric pressure (Figure 5). page 5 Linsenmeyer Attachment of Chitons Chitons could clearly attach to porous glass, wire screen, and Plexiglas with 3 mm holes. This result also argutes against suction as a major factor involved in the resistance of chitons to removal. A chiton on the 3 mm thick Plexiglas with 3 mm holes passed part of its foot through a hole and expanded it on the opposite side. providing a mechanical wedging which held the chiton to the surface. Similarly, portions of the chiton's foot penetrated the wire screen and expanded on the opposite side. When chitons were detached from smooth surfaces, a relatively insoluble mucus-like residue (hereafter "mucus") the shape of the chiton's foot remained on the surface. The chiton's resistance to removal from such smooth surfaces may be largely due to the adhesive effect of the mucus they secrete. It appears that on rough surfaces, the bonding of chitons to the surface is partially adhesive, due to mucus, and partially mechanical as the foot conforms well to surface irregularities. Furthermore, the foot appears to be the sole organ of attachment. SUMMARY 1. The lateral force required to dislodge the chitons Mopalia lignosa, Mopalia muscosa, Nuttallina californica, Katharina tunicata, and Stenoplax heathiana was directly page 6 Linsenmeyer Attachment of Chitons related to the degree of exposure to wave action in the habitat of each species. 2. The rougher the surface to which a chiton was attached, the greater the force required to remove the specimens from the surface. 3. The foot plays the major role in attachment by secreting an adhesive mucus material and by mechanically conforming to surface irregularities with its tissue. 4. Suction does not appear to play a major role in the attachment of species tested. ACKNOWLEDGMENTS I wish to express my appreciation for the help and encouragement received from the faculty and staff of Hopkins Marine Station. Special thanks to Dr. Gilmartin, my advisor, for his time and many helpful suggestions throughout this study. page 7 Linsenmeyer Attachment of Chitons LITERATURE CITED Arey, Leslie B. and W. J. Crozier 1919. The sensory responses of Chiton. Journ. exp. Zool. 29 (2): 157-260; 14 figs. (October 1919) Hoffmann, H. 1931. Amphineura und Scaphopoda. H. G. Bronn (ed.). Klassen und Ordnungen des Thier-reichs. Vol. 3. Abt. 1. page 8 Linsenmeyer Attachment of Chitons FIGURE CAPTIONS Figure 1 Device used to determine resistance to removal by a force parallel to the attachment surface. Figure 2 Device used to determine resistance to removal by a force perpendicular to the attachment surface. Figure 3 A comparison of the lateral force required to detach the five species of Polyplacophora in relation to the degree of wave action to which they are normally exposed. Bar graphs show mean and standard deviation. Figure 4 The lateral force required to detach Mopalia muscosa from various substrata. Bar graphs show mean and standard deviation. Figure 5 The vertical lifting force required to detach Mopalia muscosa from a Plexiglas plate at atmospheric pressure and at vacuums of 127 and 380 mm of mercury below atmospheric pressure. Bar graphs depict mean and standard deviation. page 9 container for weights pulley lever 2 chiton FIG V 20 em Linsenmeet container for weights pulleys — 20 cm ——1 chiton FlG. 2 Liisenmeget C . X- 237 N= 15 S.D.- 61 X-146 N= 12 5.).= 37 X=112 N- 44 5).- 49 X- 94 X- 64 N= 31 N= 8 S). = 29 5.d.- 27 3 52 Rock Exposed Rock Under Rock 46111 F16 3 Unsenneet .. ..1...r 200 3 100 X=112 N=94 S.D.-49 Glass X-144 N=2 S.D.=56 4120 Substratum FIG X=223 N=6 S.D.=69 Rock iasenmeg — 50 X-87 =13 S.D.=20 X-82 N=9 S.D.=25 380 127 Vacuum (mm of Hg) FIG 5 —, X=9. N=8 S.D.=43 nsennger