Peppard Shell Growth and Repair in the Gastropod Tegula funebralis By Margaret Caroline Peppard Shells of Tegula funebralig (A. Adams, 1854) inhabiting the intertidal areas of Mussel Point, Pacific Grove, California, are rarely found to have more than four whorls, irrespective of the size of the snail, due to heavy erosion of the upper parts. Questioning the nature of repair of erosion damage led to a consideration of the more general question of shell repair in T. funebralis. Fretter and Graham (1962) discuss shell form- ation in prosobranch molluscs, but little is understood of shell repair mechanisms. Eegla funebralis lives in what is essentially a tapered tube, closed at the small end. This is clearly seen in Figs. 1 and 2. Macroscopically, there are three layers in the shell. The thin, transparent periostracum on Mussel Point specimens is present only on the body whorl near the shell aperture, if at all. Underlying the periostracum is a black prismatic layer. These two layers are secreted only by the mantle margin. Innermost is a thick nacreous layer, white over most areas, but sometimes yellow or greenish in the upper whorls. Slides of decalcified shells embedded in paraffin show the laminar character of the nacreous matrix. Most of the specimens of Tagula funebralis from Mussel Point have shells which are conspicuously eroded. Although some of the erosion appears to be due to the radular action of Peppard predaceous snails, boring of bryozoans and polychaetes, or mechanical wear, all save a minority of T. funebralis(indi- viduals measuring 5 mm or less at the greatest basal diameter) are pitted over most of the eroded surface. Under 30x magni- fication, this damage closely resembles that caused by a fungus described as infesting shells of marine animals by Bonar (1936). Attempts were made to culture the fungus on T. funebralis on a medium of 100 ml sea water, 1 gm Caco,, 1 ml 1 M NaNo,, 1 ml 1 M KH,PO., 1.5 gm agar, 1 gm humus, 5 gm Tegula funebralig shell, finely ground, O.1% glucose, and 0.01% yeast extract. The fungus in culture shows chlamy- dospores (see Fig. 3) which differ from those on the species raised by Bonar, and also from those found by Johnson (1962) in a fungus growing on smooth jingle shells (Anomia simplex d'Orbigny) from Pivers Island beach, North Carolina. Normal shell growth in Tegula funebralis was measured for a fifteen-day period (May 14-29, 1963) on individuals ranging 13.0-27.5 mm in greatest basal diameter. Measurements were made with an ocular micrometer of growth increments on the outer lip of the aperture, secreted on top of a baseline of fingernail polish painted on the edge of the aperture at the beginning of the study. Results are shown in Table 1. The average overall addition to shell aperture was six microna per day, but growth occurred in spurts, not evenly each day. Total growth over the fifteen-day period did not measurable affect the greatest basal diameter of the shells. In order to assess the ability of Tegula funebralis to Peppard repair damage incurred to the shell, snails were operated on in various ways, inflicting different types of shell damage, as indicated in Table 2. Five individuals were operated in each way. The holes (windows) made in the shell back of the aperture were ground on an emery wheel, care being taken to keep the shell wet and cool, and the internal tissues intact. Table 2 gives the average change in each group on successive days. The range of variation within each group was not so great as to make the average irrelevant. In every case of damage to the shell aperture, growth of the damaged part proceeded faster than the growth at apertures of undamaged controls. All such operations on the apertures were repaired by the folds at the border of the mantle. The same was the case with the windows over mantle cavities. New shell mater- ial included a black prismatic layer. When the mantle margin was slit in an otherwise undamaged specimen, within two days a notch appeared in the shell aperture at the point apposed to the incision. It is not clear whether the notch was due only to lack of growth, or in part due to active resorption of shell at the point, but wehin six days the notch was repaired. Snails with openings over the visceral hump first secreted a soft, membranous layer across the inside of the hole; this later became impregnated with calcium carbonate. Successive layers, similar in appearance, were built up beneath the first layer, which bulged through the opening. After thirty days 172 Feppard (April 30-May 29, 1963), one specimen had plugged the shell window with a hard patch of white material, apparently calcium carbonate embedded in an organic matrix. The patch protruded through the opening like a bubble, and was translucent at the eperiphery, opaque in the center portion. Of three females and two males, with windows cut over the visceral hump, the females began repair sooner than the males. None of the snails with windows over the visceral hump died, al- though the gonad was frequently ruptured. On the other hand, animals wih the shell damaged by grinding a hole over the region of the heart and kidney died in all cases except one. Death was due not to the operation, but to the later rupture of the kidney or pericardial sac against the sharp edge of the opening produced by the operation. One specimen which lived an entire month with this operation failed to successfully repair the damage, for each time the soft membranous layer covering the hole became embedded with calcium carbonate, it was sloughed off through the opening. In the "windowed" animals, even where the holes penetrated yellow and green layers, I observed no secretion of yellow or green material by the mantle covering the body, nor is it secreted by the mantle margin. Secretions by other than the mantle margin were always either transparent membranous layers or white inorganic material. However, natural repair does sbow yellow or green material, particularly in eroded areas at the shell apex. Perhaps the inner layers of nacreous material are dyed by pigments secreted by the visceral hump, specifically 72 Peppard either the digestive gland or the gonad (see MoGee, 1963a). Cracked shells were bound firmly together in a solid unit within five days by a calcium carbonate-embedded membrane on the inner surface. Additional white nacreous material was laid over the outer surface of the crack within sight days where the break passed through the underside of the body whorl adjacent to the shell aperture. Summary 1. Shell erosion is caused by the activities of several animals (bryozoans and polychaetes), by mechanical wear, and by a fungus, which was cultured on agar plates. Normal shell growth, recorded over a period of fifteen days in twenty-eight animals, was intermittant, but averaged six microns per day added to the outer lip of the aperture. 3. Repairs to shells damaged mechanically, by filing the aperture, grinding holes in the body and upper whorls, and by cracking in a vise are described. Peppard Table 1 Growth Studies on the Shell of Tegula funebralis Legend: no change since last observation no observation dead Snail Greatest Addition to Shell at Aperture on Successive Day basal mm) no. diam. (mm) 1 8911 13 7060 .09 13.0 - .090 13.5 - .120 .210 - * 16.0 .030 .060 .090 .060 .030 17.0 .030 .060 17.0 .030 18.0 .030 .060 18.0 19.0 .090 20.0 .120 2.5 .090 .120 10 11 21.0 .165 12 .100 21.0 22.0 .165 100 13 .033 .132 23.0 .060 .090 .120 .150 3.C 090 3.0 .030 060.090 3.0 .030 060 18 23.0 .030 .060 .090 24.0 24.0 21 .090 24.0 .060 090.120 24.5 .060 25.0 .015 .030.120 25.0 .090.120 .030 .030 25.5 .132 .165 27. .060 27.5 .060 .120 Average growth day 506 .014 .004 .006 .004 .002 .004 008 .008 .011 O11 006 .010 .006 .004 .006 006 008 008 008 002 O11 .007 .008 175 Peppard Table 2 Repair of Shell Damage in Tegula funebralis Legend: no change since last observation no observation just visible trace of nacreous layers secreted soft, membranous layer secreted over opening calcium carbonate embedded in soft layer Caco, dead Peppard Table 2 Repair of Shell Damage in Tagula funebralis Type of Operation Repair on Successive Days After Operation (mm Added to Shell at Aperture) 11 19 — control(no opera¬ .165 .1 tion; normal growth at aperture lip recorded) 2mm notch Mantle margin slit notch filled .O66mm.240mm .3881m 2.6mm notch filed .479mm.677mm in edge of shell of notch of of of aperture filled notch notch notch notch filled filled filled filled window over Caco, 2 Caco. layers visceral hum window over heart Gaco, and kidney area window over mantle .18 .397 .198 .353 cavity added to hole edge shell cracked white Caco, (with a vise) nacreous sevretion over crack(xx) .354 shell broken at .386 .187 .221 aperture (with pliers) shell aperture .583 .333 .397 .353 ground off Pepperd Figure Captions: Fig. 1. Ventral view of Tegula funebralis, 8 mm in basal diameter; decaleified cleared in cedarwood oil, and with black prismatic layer removed Fig 2. Dorsal view of Tegula funebralis, same individual as shown in Fig. 1. Fig. 3. Fungus found on Tegula funebralis shell; part of mycelium with chlamydospores. a, shell aperture; an, anus; em, columellar muscle; et, ctenidium; e, eye; f, foot; g, gonad; h, heart; ht, head tentacle; k, kidney; me, mantle cavity; 8a, operculus; ad, esphradium x, rectum: z, chell; ge, spiral caecun; 2t. sterach. C a D