Vogelzang Abstract: Cucumaria curata and C. pseudocurata live and feed in the hydrodynamically stressful environment of exposed intertidal areas. They have been observed to actively feed in flow velocities up to 0.2 m/sec., while allowing their feeding tentacles to bend with the current as flow increases. They suspension feed with peak effectiveness at some optimum low water velocity. Consequently, it is suggested that they incorporate detrital feeding into their food-gathering behavior in order to supplement the aerosol particle capture. However, this too presents mechanical problems. The feeding strategies and mechanisms of C. curata and C. pseudocurata may depend on local water velocities within their different microhabitats, and thus help to differentiate their respective niches. Vogelzang Introduction: Cucumaria curata and C. pseudocurata are small (up to 4 cm when fully extended), dark brown holothuroids that live in exposed intertidal regions on the Central Cali- fornia coast, e.g. Yankee, Pescadero, and Cypress Points just south of Pacific Grove. These cucumbers possess ten feeding tentacles, or buccal podia, arranged radially around the mouth. The buccal podia consist of cylindrical stalks that are branched much like deer antlers and have mucous- covered papillae at the tips (see Figures 5 and 6). The cylindrical stalk is simply a 1.75 x 10 m thick tissue wall surrounding a fluid-filled lumen. This wall consists of a cuticle, epidermis, dermis, longitudinal muscle fibers, and ciliated epithelium (Brumbaugh, 1964). Within the dermis are circumferentially-arranged fibers making up the collagen sheath. The papillae are sticky protrusions to which food particles adhere. Food particles are removed when the animal retracts the entire stalk and inserts it into the mouth. Sea cucumbers are known to feed by extending their buccal podia into the current to capture suspended particles. by "shoveling" substrate into their mouths, and by picking up detritus off solid surfaces (Hyman,1955). In light of the fact that C. curata and pseudocurata are not very sturdy animals, it seems unlikely that they Vogelzang should survive in environments of strong wave exposure. This study was undertaken in order to examine these organisms behavior at various water velocities in relation to the mechanics of their buccal podia and feeding actions. The results, along with Brumbaugh's lab observations and diet estimates for C. curata (1964), and La Barbera's ophiuroid particle filtration data (1978), suggest that C. curata and pseudocurata perform both suspension and de¬ trital feeding. The actual mechanics of their feeding methods, however, cannot yet be determined without more conclusive data. Vogelzang Materials and Methods The apparatus used (see figure 1) allowed close observation of the animal and visual approximation of buccal podia angles through the glass tube. Flow was manually controlled at the spigot and directly measured with a 500ml graduated cylinder and stop watch. Average velocity was calculated from flow by the equation: flow (ml/s)/area(cm2)-velocity (cm/s) where area is the cross-sectional area of the Cucumaria subtracted from that of the tube. Active animals were fixed in glutaraldehyde while pressure was applied to the body in order to maintain extension of the B.P. papillae. These fixed animals were then placed in, first sea water, second distilled water, and gradually dried in acetone, dry acetone and the C0» critical point dryer. Dried specimens were gold-coated for 1.5 minutes. and examined under the SEM. Measurements of B.P. papillae diameter, surface area, and stalk length were taken from the scanning electron micrographs. By dissection of cucumbers relaxed in 50/50 MgCl, and sea water, gut contents were obtained and visually analyzed under 100X and 400X magnification for food particle sizes. B.P. were amputated from active animals and embedded in 20% gum arabic with 10% formalin. Sections were cut in freezing microtome. The stalks' inner and outer radii buccal podia Vogelzang (see figure 3) were measured with a scaled eyepiece under 100X magnification. Center of area for the B.P. stalks was calculated from measurements of center of mass of cardboard models The models were hung sequentially from thread at various points, and the intersection of traced lines of gravity was assumed to be the center of mass (Alexander, 1968). Vogelzang Results: Drag vs. Velocity The pressure drag, Da, and frictional drag, Dp, forces were calculated by the formulae Dp - 2 Cp' S Ufe (Vogel, 1981), and Dp - 2 Cp S U60 (Hoerner, 1965), where the density of sea water,o, is 1.024 x 10 kg/m, and U is the water velocity. The surface area, S, is .252 x lengthx width of the buccal podia stalk where .252 is the ratio ig for the Cucumaria in the SEM (Figure 5). Cp', the useful drag coefficient, was obtained from the equation '- Cp sin, which accounts for the change in angle of the buccal podia in relation to the flow as water velocity increases (Hoerner, 1965). The angle was estimated visually, and Cp and Cp were found using standard drag coefficient vs. Reynolds Number plots for cylindrical objects (Vogel, 1981 and Hoerner, 1965). Reynolds Numbers were calculated using the Re - Pea A in which u 2 1.1 x 10 2 kg/m-s, the viscosity of sea water at 15°0; the diameter, d, of the buccal podia stalk was estimated from that of the Cucumaria shown in Figure 5. The resulting graphs from plots of drag vs. velocity (see Figure 2) show an increase in drag with increasing velocity. The calculated shifts in these curves between velocities of .07 -.19 cm/s are due to an observed step Vogelzang change in angle of the buccal podia and to peculiarities of the relationship between Cpand Re at these velocities. Stress To determine the stress due to pressure drag that the wall material of a buccal podia stalk might encounter, the stalk was assumed to behave as a cylindrical cantilever, and the formula G - M was used (Wainwright, 1976). M, the applied bending moment, equals DpX L, where D. is the pressure drag (a typical value of 10 kg-m/s as calculated for an angle of 60°, surface area of 1.32 x 10 m and velocity of .2 m/s) and L is the length from the base to the center of mass (as determined with card¬ board models and a ratio of L/D 2 3.64. See Figure 3). is the average radius of material in the stalk Y - (see Fig. 3), and I - is the second moment of area. The stress (long before breakage) due to pressure drag for the experimental angle of 60° and velocity of 20 cm/s is between 10 and 10“ N/m. The stress for the (100u-thick) collagen sheath in the buccal podia stalk, at an angle of 90° and field velocities up to 5 - 16 m/s, results in values on the order of 10° and 102 N/m2. The stress due to frictional drag was calculated (D/area) to be 10'-10- N/m' for the collagen sheath in a buccal podia stalk at an angle of 180° in field velocities up to 5 - 16 m/s. Vogelzang Mode of Particle Capture The aerosol particle filtration theory offers five mechanisms by which particles may be trapped. (See Figure 4) This theory involves a filter system of widely spaced sticky fibers, in this case, papillae. This theory replaces the classical ideas of sieving, wherein the captured par¬ ticles must be of larger diameter than the spacings of sieving elements. Four of the mechanisms were examined in relation to C. curata and C. pseudocurata particle capture. The fifth mechanism involves electrostatic attraction and does not significantly affect particle capture in seawater due to an abundance of ions.) (Rubenstein and Koehl, 1977) The index of direct interception as a method of particle - dp/ capture was determined using the equation Non 1.689. (Rubenstein and Koehl, 1977, after Pich, 1966) dp, the diameter of particles captured, is 25 (diatom sizes measured visually from gut contents); and da, the "fiber" diameter, is 14.8 (papillae measured from the SEM. Figure 6). The index of motile particle dispersion (Rubenstein and Koehl,1971, after Pich,“ -— - 4.14 x 10 %. K, Boltzmann's Constant, mf 3rudp Udp is 1.38 x 10 22 J/0K; T - 288°K, the absolute temperature of the sea water; + - 1.1 x 102 Ns/m, viscosity; dp 2.5 x 10 2 m, particle diameter; dp - 1.48 x 10 2m, fiber diameter; and U -.25 m/s, typical water velocity. The index of gravitational deposition 10 Vogelzang = dp2 g (ep - P) /18 uU - 2.48 x 10*2 (Rubenstein Na and Koehl, 1977 after Chen, 1955) to 1.86 x 10 if the difference in density between particle (diatoms) and fluid Rubenstein and Koehl, 19 14 aftcr medium, (ep - Pw), is 20 (Woodley, 1967) to 150 kg/m (Hutchinson, 1967) respectively, where g is gravity - 9.807 m/s. The index of inertial impaction Nyp - [(ep - Pw) dp2 U/ 18 udp (Rubenstein and Koehl, 1977 after Fuchs, 1964) - 1.07 x 10 6 to 8.0 x 10, again if (ep - Pw) - 20 to 150 kg/m2. Since Npp is considerably larger than either NIE: NGE; or Nyp; direct interception is the most important mode of particle capture for C. curata and C. pseudocurata amongst the aerosol suspension methods. Boundary Layer The boundary layer, a function of viscosity, is an area of retarded flow adjacent to the surface of an object in the flow. A rough estimate of the boundary layer thickness (as if the buccal podia did not affect the flow around the cucumbers' bodies) was calculated from the equation 6- 586 (Vogel, 1981), where 4 is the boundary layer thickness, x (extended buccal podia measured under dissecting scope and values applied to similar size animals) is the distance from the base of the stalk to the tip, u is the viscosity of 15°0 sea water, o is the density of sea water, and U (values at which buccal 11 Vogelzang podia were 90 - 120° relative to current) is the water velocity, According to calculations using experimental velocity data, the boundary layer thickness of 10 - 10 2 m would not be great enough to include the buccal podia at angles up to 135° relative to the current (which appears to be the maximum angle under experimental conditions). Sticky Surface Area The percent surface area of each buccal podia stalk covered by mucous-covered papillae was calculated by first figuring the surface area of the papillae and multiplying by the number of papillae clusters as they appear on a single stalk in the SEM (Figure 5), and finally dividing by the Kpap = percent sticky surface stalk surface area: pap stalk area of E 0.05%. 12 Vogelzang Discussion C. curata and pseudocurata are found associated with coralline algae in tide pools and clinging to wave-washed rocks in the mid to low intertidal zones of exposed coastal areas. Due to the intensity of wave action at these sites, it is not feasible to observe the cucumbers at high tide. However, the cucumbers are not observed to actively extend their buccal podia at low tide, indicating that they feed during high tides. Yet high tide, with its flow velocities of 5 to 16 m/s (Vogel, 1981), seems an unlikely situation for Cucumaria to be effectively feeding. How do these animals feed? According to Brumbaugh's estimates (1964), planktonic diatoms comprise between 15 & 50% of the C. curata diet, while detritus and sessile diatoms make up 10 to 708 of the food source over a period of one year. Thus these animals feed by sampling both the water moving by them and the surface to which they are attached. What requirements do these modes of feeding place on the structure and materials of buccal podia? High tide flow velocities impose drag forces on an extended B.P. stalk. Overall drag force is a combination of pressure drag and frictional drag. The maximum value of 10"1.10° N for pressure drag is calculated here for B.P. at 90 in 5-16m/s current. This is unrealistic in that experiments show that flow velocities as low as 0.3-0.8m/S create enough stress for the B.P. to fold backward to 135 with the current; and only up to velocities of 0.2- 0.3 m/s can the animals maintain their B.P. at angles of 13 Vogelzang 90. For the folded back B.P., pressure drag no longer exerts any substantial force on the B.P. stalk. As stated previously, experiments show that B.P. bend back to 135 at velocities of 0.3 m/s; therefore B.P. would certainly be folded completely back (180) at currents of 5-16 m/s and probably as low as im/s. Here, friction drag, 10-3 to 10-2 N, is calculated for B.P. 5-16 m/s current at 180. Both pressure and friction drags impose stresses on the buccal podia stalk. If it is assumed that the force is resisted solely by the collagen sheath, the imposed stress due to pressure drag would be 10°-109 N/m2 for the unrealistic situation of B.P. holding up at 90' in 5-16m/s current. Not only is this situation unlikely, but it is probably impossible. The collagen of rat tail tendon breaks at a stress of 10'N/m (Wainwright, et al, 1976). Therefore the B.P. would be torn apart at the calculated stress due to pressure drag of 10°-10? N/m2. However, the stress due to frictional drag as calculated for 180' B.P. even in 5-16 m/s current, is 10“-102 N/m2, well below the breaking stress values. Among the mechanisms of suspension-feeding outlined by aerosol particle filtration theory (Rubenstein & Koehl,1977) direct interception is the most important, as indicated by its high relative index (see results). The index of direct interception is independent of water velocity, but 14 Vogelzang suspension-feeding effectiveness by this method (defined as the number of particles captured /time) does depend on the total volume of particles intercepted by the buccal podia's sticky surface area; and therefore is a function of velocity. Increased water velocity results in the following gains in suspension-feeding effectiveness: (1) A large volume of fluid flowing past the buccal podia per time. (2) A thinner boundary layer adjacent to the buccal podia. The decreased velocity in a boundary layer would decrease the volume of water intercepted. (3) An increased index of inertial impaction, thus slightly increasing the suspension-feeding effectiveness. In contrast, increased water velocity simultaneously diminishes suspension-feeding effectiveness as follows: (1) Particle capture indices for motile-particle disper- sion and gravitational deposition decrease. (2) Drag force, and therefore stresses are increased which cause the B.P. to fold back, decreasing the exposure of sticky surface area to flow, and consequently reducing particle capture. It seems likely that there exists some optimum water velocity at which these cucumbers suspension-feed most effectively. Although exact figures cannot be calculated at this point, the optimum velocity is likely to be below 1m/s since 180' B.P. bending occurs above this velocity and reduces suspension-feeding effectiveness by minimizing 15 Vogelzang exposure of sticky surface area to flow. Considering the suggested low range of water velocities these species require in order to be effective suspension¬ feeders, it is unlikely, according to aerosol particle filtration theory, that C.curata and C.pseudocurata are exclusively suspension- feeders. This conclusion is supported by a comparison of these cucumbers with two species of ophiuroids that are thought to be strictly suspension-feeders (Hyman, 1955). The calculated particle capture indices for C.curata and pseudocurata are in rough agreement with those for Ophiopholis aculeata (La Barbera, 1978) and Ophiothrix fragilis (Rubenstein and Koehl,1977). (see figure 8:) However, the cucumbers' mucous-covered papillae represents only -0.05% of the surface area of each B.P. stalk. Ophiuroid arms extend a much larger relative surface area into the water column. Unless the cucumbers encounter a correspondingly greater volume of particles in their fast flow regimes, the ophiuroids are more effective suspension feeders. This analysis implies that suspension particle capture is unlikely as an exclusive feeding mechanism for C.curata and pseudocurata. Inevitably, these cucumbers need an additional feeding mechanism. Based on behavioral observations, the only likely alternative is detrital feeding. Brumbaugh reports C.curata performing detrital-feeding 16 Vogelzang behavior under the dissecting scope (Brumbaugh, 1964); the cucumber sweeps its feeding tentacles along solid surfaces, picks up settled material, and ingests the detritus indiscriminately. Though much of the ingested material non-nutritive, Brumbaugh states that this is likely to be the primary food-catching method. However, as with suspension-feeding, the B.P. must be extended when detrital feeding and thus experience frictional and pressure forces as they are swept along the substratum. These forces create the problem of folding the B.P. back at water velocities as low as 0.3 m/s experimentally. On the basis of the observations made here, it seems likely that fluid dynamic forces thus render the feeding tentacles less functional for effective suspension feeding and, to a greater extent, non-functional for detrital feeding at field velocities. Yet these intertidal Cucumaria have been reported to use a combination of suspension and detrital feeding. How might this paradox be explained ? It is possible that the conclusions reached here will need to be modified when careful measurements are made of the feeding behavior and flow in intertidal microhabitats for example, local flow velocities around the B.P. crown may be affected by the cucumbers' behavior: clustering with one another, wedging themselves into crevices, living at the base of the coralline alga Calliathron and even crawling under the encrusting coralline algae. 17 Vogelzang C.curata and pseudocurata exhibit slightly different behaviors and occupy slightly different habitats in the field. Specifically, C. curata has 10 evenly arranged buccal podia and is generally larger in size than C. pseudocurata. C.curata lives isolated, in pairs, or in small groups located within vertical rock crevices in wave-exposed zones. In contrast, C.pseudocurata tends to aggregate in clusters of numerous individuals on the encrusting coralline algae at the bottoms of protected tidepools. This smaller species has two ventral B.P. which are reduced in size compared with its other eight (Collison, 1983). These differences may possibly play a role in affecting feeding behavior and local flow in each species' microhabitat. Further investigation is necessary in order to determine niche differentiation between C.curata and C.pseudocurata, especially in relation to feeding mechanisms. One intriguing observation (with potential for closer examination) was that of the cucumber's apparent stream¬ lining behavior, when the anterior end was directed into high velocity flow of more than 0.8m/s (see fig. 7). When the animal's anterior end oriented away from the current, it still humped up the anterior end. More detailed investigation is required to analyze the effectof this second shape on flow forces. This study has examined aspects of the mechanics of feeding by C.curata and C.pseudocurata on wave-swept shores. Present understanding of buccal podia mechanics Vogelzang does not shed light on the problems of feeding under conditions of fast flow. Resultant stress prevents full and effective extension of the B.P. More measurements and observations of behavior need to be made in the field to better correlate laboratory results to various natural conditions of microhabitat such as high tides during the day and night. Vogelzang Many thanks go to Mark Denny for providing his direction, motivation, and encouragement. Thanks also to Freya Sommer for her patient listening and support and to Magruder for his teaching and technical assistance. To the dwellers of 629 Jewell, my warmest thank you for late night cookies and laughter. 20 Vogelzang References Alexander, 1968. Animal mechanics. University of Washington Press, Seattle. Brumbaugh, J. H. 1964. The anatomy, diet, and tentacular feeding mechanism of the dendrochirote holothurian Cucumaria curata Cowles 1907. Ph.D. diss. Stanford. Collison, J. 1983. Establishment of niche differences between Cucumaria curata and C. pseudocurata. Unpublished. Hoerner. S. F. 1965. Fluid-dynamic drag. P.O. Box 342, Bricktown, NJ. Hutchinson, G. E. 1967. A treatise on limnology. Vol. 2. Introduction to lake biology and the limnoplankton. Wiley, New York. Hyman, L. H. 1955. The invertebrates: echinodermata. Vol. 4. McGraw Hill Book Co., Inc., New York. La Barbera, M. 1978. Particle capture by a Pacific brittle star: experimental test of the aerosol suspension feeding model. Science 201:1147-1149. Rubenstein, D. I. and M. A. R. Koehl. 1977. The mechanisms of filter feeding: some theoretical considerations. Am. Nat. 111:981-994. Vogel, S. 1981. Life in moving fluids. The physical biology of flow. Willard Grant Press, Boston. Wainwright, S. A., W. D. Biggs, J. D. Currey, and J. M. Gosline. 1976. Mechanical design in organisms. Edward Arnold Ltd., London, UK. Woodley, J. D. 1967. Problems in the opiuroid water- vascular system. Symp. Zool. Soc. London 20:75-104, Vogelzang Figure Legends 1. Flow apparatus 2. Graphs of Drag vs. Velocity (on four separate pages) 3. Schematic representation of buccal podia stalk cross section and buccal podia models for determination of center of area 4. Aerosol suspension particle capture methods A. Sieving B. Direct interception C. Inertial impaction D. Motile particle dispersion E. Gravitational deposition 5. Scanning Electron Micrograph of buccal podia crown 6. SEM of papillae 7. Streamlining 8. Indices of aerosol particle filtration C S S (C C C VELOCITY 20 A 90° Cucumber Length .8 cm Width.2 cm 40o-2 60 m/s VELOCITY 1O A 90 Cucumber Length 1.5 cm Width .35 cm 200-2 30 m/s 40 VELOCITY 20 490 Cucumber Length 2.0 cm Width .2 cm -260 m/ 80 C 9 VELOCITY 1O A 90° Cucumber Length I.O cm Width .3 cm 20 0-2 30 m/s 40 Cross-sectioned Buccal Podia Stalk 0. ol 8 L/d-3.64 L= dx3.64 A —— 2 — — W C Streamlining — . .. posterior anterior anterior posterior C C ... M Nor M Ophiopholis aculeata Ophiothrix fragilis Cucumaria curata & pseudocurata 0°-10 10 10 03 10-2 0-2 -10 10 0-2-0—4 10-40-5 05 100 0-0 10