Sandy-lobed Ascidians Mulroy, W. Abstract The similarities in external morphology and distribution have made distinction between Aplidium arenatum, Aplidium propinquum and Synoicum parfustis difficult in the field and laboratory. Quantitative differences have been found between the species in: height and area of the lobe top; the shape, color, and clustering of lobes; the number of systems per lobe; the number of zooids per lobe; the number of zooids per system; zooid size the number of rows of stigmata; the number of developing embryos in the atrium; and habitat. 1 Sandy-lobed Ascidians Mulroy, W. Several species of compound ascidians in which the colonies form sandy lobes occur near the Hopkins Marine Station at Pacific Grove, California. They are so similar that species identification in the field and even in the laboratory may be difficult. The original descriptions of these species bring out more similarities between them then differences. Van Name (1945) described Aplidium arenatum as forming irregularly club-shaped lobes thickly encrusted and impregnated with sand, the top of the lobes in a cluster fitting together to form an even surface. He found that lobe height reached 25 mm but was usually less than 5mm. Aplidium propinquum was described by Van Name (1945) as irregularly club-shaped lobes similar to A. arenatum, although reaching a greater height. Ritter and Forsyth (1917) described the external appearance of Synoicum parfustis as sand-encrusted, club-shaped masses having a peduncle twice as long as the rounded head. The height of an individual lobe reaching 6 cm. In addition to the similarity in appearance, all three species are distributed in the low intertidal zone. The zooids all have three body regions and form systems in a transparent, common test. Positive identification requires studying such details of zooid structure as the contour of the stomach wall and the number of rows of stigmata in the pharynx. The objective of this study was to describe the range in variation for each species, both in external characteristics of the lobes and in zooid characteristics, to make identification easier in the field and in the laborator) Laboratory studies Methods: The methodology was simple. All sand encrusted lobes that looked Mulroy, W. Sandy-lobed Ascidians different were collected from the intertidal zone. The organisms were then placed in bowls of seawater, relaxed overnight with menthol, then fixed in 10% formalin for 48 hours. Only well-expanded zooids in which all rows of stigmata were clearly visible were used for detailed study. Measurements of zooids and lobes were made with a Vernier caliper. Results: A quantitative measure of shape of the lobes is provided by plotting the height of a lobe against the area of the lobe top (greatest width of the lobe multiplied by the greatest thickness) for many lobes of each species, The results are shown in Figurel. and original data are tabulated in Appendix A. Aplidium arenatum has the smallest and thinnest lobes of the three species. The largest lobe measured 35 mm in length. A. propinquum lobes reach 50 mm in length and are more plump than those of A. arenatum. Synoicum parfustis lobes reach the greatest size. Note the overlap in size and shape of the three species in the smaller lobe size range. Clearly, other characteristics must be studied for identification. Another value showing some differences between species is the number of systems per lobe (Figure 2.). A small area at the top of the lobe allows only a small number of systems. On the graph note that the 7+ category of the number of systems includes both A. arenatum whose maximum value was 8 systems in a lobe and Synoicum parfustis which commonly had over 12 systems in a large lobe. In the area range of 50-200 mm’, A. propinquum and Synoicum parfustis have similar values. Figure 3. compares area of the lobe top with the total number of zooids in the lobe. A. arenatum never had more than 27 zooids in the lobes studied. In Synoicum parfustis large lobes have well over 100 zooids. Sandy-lobed Ascidians Mulroy, W. Although A. propinquum lobes do not have as many zooids as large lobes of Synoicum parfustis, zooids are more densely packed in a given area. Figure 4. displays the number of zooids per system. Aplidium arenatum averaged 5.4 zooids per system, and lobes containing one zooid were unique to A. arenatum. Most systems of Synoicum parfustis had between 6-8 zooids. In A. propinquum, with an average of 10.8 zooids per system and as many as 15 zooids in one system, the individuals are more densely packed than in the other species. The variation in size and shape of lobes of each species is compared in Figure 5. From observations made on each species the external appearance of the lobes in clusters is as follows: Aplidium arenatum is a small, brown, slender lobe occurring in tightly packed clusters. The small size, heavy sand-encrustation and flat lobe tops give the cluster top the appearance of a continuous mound. Sand not only encrusts the outside of the common test but is loosely spread throughout the inside also. Aplidium propinquum forms more plump lobes and larger clusters than A. arenatum. The lobes range up to 50 mm in height although 30 mm is more common. The heavy sand encrustation gives way to the transparent tunic at the bright orange-red, common cloacal aperture, giving the system opening a distinctive crater-like appearance. Synoicum parfustis described by Abbott and Newberry (1980) as clusters of orange, tear-drop shaped lobes are unmistakable when they attain this shape in well-developed lobes. Smaller lobes are not as distinctively shaped; however the heads are still more rounded than flat-topped, and the bright orange systems can be seen in life. Sandy-lobed Ascidians Mulroy, W. The growth of the lobes of each species is also reflected in Figure 5. A. arenatum, A. propinquum, and Synoicum parfustis all form new lobes from the bases of existing lobes. All three species can add to the area of a lobe top by increasing the number of systems in the same lobe head. A. arenatum and A. propinquum have a third method of growth, branching of a lobe. A new system may form and split the lobe top forming two heads. This lobe branching adds to the tight clustering of the colonies, Figure 6. compares the zooids of each species. The drawings, taken from Van Name (1945), show the number of rows of stigmata and the contour of the stomach wall. It is the zooids which give the lobes their characteristic color. The oral and atrial siphons of A. arenatum are brown. The brown coloring fades to a transparent thorax. In A. propinquum the thorax is a bright orange-red color, and in Synoicum parfustis it is bright orange. Figure 7. displays the variation found in the number of rows of stigmata for each species. A. arenatum always has 5 rows of stigmata. A. propinquum has 15-21 rows of stigmata, while Synoicum parfustis has 10-16 rows. All counts were made on well-expanded zooids in which the number of rows could be counted accurately. te zige ahe In Figure 8., species are compared with respect tozooid. Since the length of the post-abdomen is too variable, the combined length of the thorax and abdomen was used as a measure of size in well-expanded zooids. A. arenatum was the smallest, maximum length 6 mm. The largest zooid of A. propinquum is 9 mm. Synoicum parfustis has the largest zooids with some reaching over 12 mm in length of thorax and abdomen combined. Figure 9. shows the number of developing embryos found in brooding zooids. A. arenatum and A. propinquum showed comparable numbers of developing Sandy-lobed Ascidians Mulroy, W. embryos in the atrium. Synoicum parfustis has the greatest capacity for brooding embryos of the three species by carrying up to 13 developing embryos. Field studies The second phase of the project was conducted to see if any distributional differences could be seen between the three species. Methods: From collection of specimens, the general area where lobes occurred was known. Fourteen areas were then selected for study on the basis of surf exposure, depth, and the presence of at least one of the three ascidian species being studied. Abundance and distribution of each species was observed in each area from the zero tide level to the ocean floor. Each area was divided into three ranges of depth below the zero tide level: +0.25 to -0.25 m,-0.25 to -0.75m and -0.75 to -1.0 meters. The zero tide level was determined using a meter stick and the Monterey Bay tide charts and measuring the zero tide level from the height of the low tide. The abundance of each species in each area and depth was measured on a scale of 0 (species not present), 1 (species present), 2 (species present in good amount), or 3 (species present in large amount). Results: General patterns of distribution are shown in Figure 10. Synoicum parfustis occurs lower down in the areas studied, A. arenatum and A. propinquum higher up. A. arenatum was not as abundant as either Synoicum parfustis or A. propinquum. Looking at Table 1., the environments for Figure 10., further information is revealed about distribution. Synoicum parfustis Sandy-lobed Ascidians Mulroy, W. grew best in protected areas of moderate surf and on horizontal surfaces in the lower portion of the depth studied. A. propinquum grew best in exposed areas with horizontal or vertical surfaces, while A. arenatum grew best in exposed areas on horizontal surfaces. Discussion: From the studies on distribution a logical question follows: what are the advantages of being a sandy-lobed, colonial tunicate? One advantage is in space competition. These compound ascidians are benthic organisms. By having only a stalk attached to the substratum most of the tunicate's mass is out in the free-flowing water. The lobe shape allows more individuals per given area of substratum, and prevents the organism from being as easily overgrown by flat colonial tunicates or sponge as an encrusting colony might be. Since the tunicates are filter-feeders, a lobe sticking out from the substratum may get better circulation of water and thus more food. Finally, the hard sand encrustation protects the tunic from abrasion in the surf as well as discouraging predators. With these advantages in gare mind, the distribution of the three species can, be explained. The distribution of Aplidium arenatum and A. propinquum higher up and in heavier surf as compared to Synoicum parfustis appears to be related to the morphological differences between the species. A. arenatum and A. propinquum form tightly packed clusters; the lobes are smaller than in Synoicum parfustis and are more heavily sand encrusted. These characteristics provide more protection from the heavyy surf and from desiccation at low tide. Synoicum parfustis on the other hand, form loose clustering colonies which doeé not allow the individual lobes to support each other in the heavy surf or to retain water in the cluster at low tide. The large size 7. Sandy-lobed Ascidians Mulroy, W. of the lobes also would be a disadvantage in heavy surf by putting more stress on the stalk than a smaller lobe would be subjected to. For these reasons Synoicum is better suited for moderate surf and greater depth below the zero tide level. dgeme Acknow I woull like to thank Don Abbott for his enthusiasm and guidance which made this project so enjoyable. Special thanks goes to Keith Kohatsu, the best T.A. ever. And finally my gratitude must be expressed to all the students in the spring class, the faculty and the rest of the staff who make Hopkins such a special place. 10. Literature Cited Abbott, D.P. 1975. Phylum Chordata: Introduction and Urochordata, pp 638-625. In Smith, R.I..and S.F. Carlton., Light's Manual: Intertidal invertebrates of central California coast. Univ.Calif. Press. 716 p. Abbott, D.P. and A.T. Newberry.1980. Urochordata: The Tunicates. pp. 177-226, pls. 57-67. In Morris, R.H., D.P. Abbott and E.C. Haderlie, eds., Intertidal Invertebrates of California. Stanford University Press. 690 pp. 200 pls. Jackson, J.B.C. 1979. Morphological strategies of sessile animals, pp 499-503. In G. Larwood and B.R. Rosen, eds., Biology and Systematics of colonial organisms, Academic Press, 589 pp. Ritter, W.E. and R.A. Forsyth 1917. Ascidians of the littoral zone of Southern California. Univ. of Calif. Publ. Zool. 16:439-512, pls. 38-46. Van Name, W.G. 1945. The North and South American Ascidians. Bull. Amer. Mus. Nat. Hist. 84:1-476, pls. 1-31. 11. Figure Captions Figure 1. Area of the lobe top vs. height of the lobe for three ascidian species. Numbers above bars show number of lobes involved. Figure 2. Area of the lobe top vs. the number of systems per lobe for three ascidian species. Numbers above bars show number of lobes involved. Figure 3. Area of lobe top vs. the number of zooids per lobe for three ascidian species. Numbers above bars show number of lobes involved. Figure 4. Variation in number of zooids per system for three ascidian species. Numbers above bars show number of systems involved. lobes Figure 5. A comparison of lobe shape for,up to 50 mm in height for three ascidian species. Figure 6. A comparison of zooid structure taken from Van Name (1945). Variation in the number of rows of stigmata per zooid for Figure 7. three ascidian species. Figure 8. Variationg in combined length of thorax and abdomen for three ascidian species. Figure 9. Numbers of developing embryos in the atrium in brooding zooids of three ascidian species. Figure 10. Distribution in depth below the zero tide level vs. the environment studied. 0 le Captions Table 1. Environments (A-N) for Figure 10. Table 2. Summary: Characteristics useful for separating three species are summarized in the following table. 12. e Table 1 Environments for Figure 10 I. Exposed areas (heavy surf) AREA VERTICAL SURFACE HORI ZONTAL SURFACE -0- -O- -0- -O- -O- -0- -0- -0- -0- -0- -0- -0- II, Protected area (moderate surf) VERTICAL SURFACE HORI ZONTAL SURFACE AREA -O- -O- -O- -O- -O- -0- -0- -O- -O- +-prasa O - alan BOTH SURFACES -0- -0- -O- -0- -0- -0- BOTH SURFACES -0- 13. Table 2 Aplidium arenatum Brown 1. Lobe color Heavy 2. Sand encrustation Flat 3. Lobe top 4. Clustering of colony. Tight 1. Lobe formation New lobe growth by: at base of another lobe. 2.Splitting of one lobe to form two. 5. Lobe shape Flat 35 mm 6. Maximum height of lobes. 7. Visibility of systems Difficult to see. in vivo. 1-8 8. Number of systems/lobe 1-27 9. Number of zooids/lobe (range) sastem 5.4 10.Number of zooids/lebe (means. 11.Number of rows of stigmata 5 12.Combined length of zooid abdomen and thorax 1-6 mm 1-5 13.Number of developing embryos in atrium Heavy surf 14.Distribution 14. Synoicum parfustis propinquum Orange Orange-red Heavy Moderate Flat Rounded Tight Separate 1.Lobe formation 1.Lobe formation at base of at base of anothe another lobe. lobe. 2.Splitting of one lobe to form two. Rounded Flat 60 mm 50 mm Orange-red, crater-/Orange. Clear like 1-6 1-15 5-124+ 8-62 6.6 10.8 10-16 15-21 2-12 mm 2-9 mm 1-14 1-6 Heavy surf Moderate surf g gegrg L — — R — — — — +Gt v-O2 68-9 1 -0 ) HH 8 O I 6 GEN fig. 1. 15 — 10 9 R — +1 9-9 v-2 2- 1807/SWZISAS ZO ON — — . L fig. 2. 6 — — — +OV 62-08 61-0 1801/SGOOZ DO ON — fig. 3. 6) 0) 0 LL 8 fig. 4. Aplidium arenatum N:24 LT L A. propinquum N. 29 I 4 2 E Synoicum 8 parfustis 81 N: 34 4 3 LL 4 4 8 O 2 NO. OF ZOOIL S/ SYSTEM 1 + J a I S L 10 N Q I fia 0 S C S — — S — S S S e fig. ( 2 kig. 7 139 40. Aplidium arenatum 70. N- 139 — 0 20 19 O A. propinquum NO N= 65 4 O 10 9 Synoicum parfustis 304 N-110 19 20 0 L 20 15 NO. OF ROWS OF STIGMATA 40 20- 20 O 101 L 30 2 20 10 22 fig. 8 42 Aplidium arenatum 28 26 Nz104 6 2 — — 12 16 2 A. propinquum 21 N=62 17 8 2 2 a - L 2 10 2 27 Synoicum parfustis N=105 2 12 11 10 9 2 12 4 15 8 9 10 22456 LENGTH OF THORAX + ABDOMEN (mm.) 20 12: 41 0 O 12 L 8 D 2 41 8 4 20 22 fig. 9. Aplidium arenatur N60 4 14 A. propinquum N=45 263 et preingn L 10 Synoicum parfustis N-66 5 2 L L NO. OF DEV. 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