ADDITIONAL INFORMATION, IF ANY, CONCERNING AUTHORS, ADDRESS, TITLE, OR CITATION DATA PLEASE TYPE ABSTRACT DOUBLE SPACED BELOW WALKER, CATHÉRINE GENE. (Hopkins Marine Station of Stanford University, Pacific Grove, California, USA.) Studies on the jaw, digestive system, and Goelomic derivatives in representatives of the genus Acmaea (Mollusca: Gastropoda: Prosobranchia) Veliger . -- Placement and extent of the radula sac, the form of the jaw, the arrangement of the salivary glands, the looping of the gut, and the interconnections of coelomic derivatives are described and illustrated for Acmaea asmi, A. digitalis, A. limatula, A. pelta, A. scutun, A. scabra. The ratio of radula length/shell length is nearly 2 in A. scutum and 1-1.5 in other Acmaea species. Jaws vary in shape; the hard anterior band in A. pelta, A. insessa, and Lottia gigantea bears an anterior median tooth. Salivary glands vary between species in length of ducts, size and position of glands. The pattern of gut loops hows minor but consistent differences between Acmaea species. Development of the Fadult pattern of loops is essentially completed before animals reach a shell length of 4 mm. The gut tends to be thicker and heavier in species feeding on larger erect or encrusting algae (A. pelta, A. limatulà, A. scutum) than in species feeding on films of microscopic algae (47 digitalis, A. Scabra, A. asmi) Limpets from the high intertidal zone (A. digitalisaand A. scabra) have relatively larger right and left kidneys than do Acmaea species from lower levels. PLEASE DO NOT TYPE BELOW THIS LINE 107 Studies on the Jaw, Digestive System, and Coelomic Derivatives in Representatives of the Genus Acmaea (Mollusca: Gastropoda: Prosobranchia) By Catherine Gene Walker Hopkins Marine Station of Stanford University, Pacific Grove, California (13 Text figures; 1 Table) Previous and contemporary workers have noted some dietary differences between California species of Acmaea (e.g., Test, 1945, 1946; Craig, 1966; Eaton, 1966), and have noted some species differences in the radula and radula strap (e.g., Test, 1945, 1946; Fritchman, 1960, 1961), which also may be related to diet. No similar attempts have been made to compare the digestive tracts or jaws of California limpets, though these too might be expected to show some variation possibly related to food habits. In the present work, the gross morphology of the gut has been compared in the following species: A. asmi (Middendorf, 1849); A. digitalis Eschscholtz, 1833; A. limatula Carpenter, 1864; A. pelta Eschscholtz, 1833; A. scabra (Gould, 1846); A. scutum Eschscholtz, 1833: The jaws have been studied in these six species and also in A. insessa (Hinds, 1842), A. paradigitalis Fritchman, 1960, and Lottia gigantea Sowerby, 1843. (*sec tstriste ) 5 106 Catherine Gene Walker -2- Additional observations have been made on the development of the digestive tract in very young Acmaea, and on the interconnection of the coelomic cavities in the six species. METHODS All specimens examined were collected on Mussel Point, near the Hopkins Marine Station, Pacific Grove, California, and at Point Pinos and Point Joe nearby on the Monterey Peninsula, during April and May, 1966. Dissections were made on animals killed in 95% alcohol. Since this type of preservation, making tissues more pliable, introduced distortion of certain structures, comparisons were made with fresh material also. Injection of Ward's latex solution was used to determine placement of gland ducts and intestinal loops. Starvation of the animals for 3 - 5 days before injection proved helpful in clearing the digestive tract, though injection of thee entire tract was never successful. GENERAL DESCRIPTION OF THE DIGESTIVE SYSTEM The gross anatomy of the digestive tract of the Acmaea species studied (Figures 1 and 2) is similar to that of Acmaea virginea (Muller) as described by Fretter and Graham (1962; pp. 149-239, 477-509), and Lottia gigantea (see Fisher, 1904). The mouth opens ventrally on when the head, isinanaturelposttionor whenthelimpet ie The lips completely encircle the elliptical opening with a wide wrinkled border. The mouth leads Catherine Gene Walker -3- into the buccal or oral cavity which contains a series of pouches, grooves, the radula, and the naw. The radula sac begins just behind the buccal mass and cavity. Passing straight back over the head muscles, it enters the anterior side of the visceral mass. It passes between the intestinal loops and stomach, curves to the right and forms a loop surrounded by the digestive gland, and extends anteriorly again, rising up over the top of the gland where often it can be seen when the shell and dorsal body wall are removed. Reentering the head cavity at the same point at which it left, the sac ends in a fleshy knob or caecum which secretes the radula (Fretter and Graham, 1962,p. 173; Runham, 1963). The jaw, a pliable chitinous structure, is posi¬ tioned anteriodorsal to the oral cavity (Figures 2 and 6). It protects the upper lip from the scraping move¬ ment of the radula teeth, and prevents food from escaping the buccal cavity (Fretter and Graham, 1962, p. 168). In Acmaea and Lottia gigantea there is a single symme- trical jaw, with four wingtlike extensions that are opaque, white, and fairly flexible. The smaller anterior wings overlap the posterior wings on the dorsal side. Muscles attached to the bases of the wings run to the body wall and buccal mass. Across the anterior mid¬ portion of the jaw runs a harder band, which becomes reddish-brown in older animals. 110 Catherine Gene Walker -4- Behind the oral cavity lie the pharynx dilation, and the esophagus. The latter extends backward to enter the visceral mass slightly to the left of the midline. Curving to the right it passes upward through the digestive gland and ends just under the dorsal surface of the gland inna mid-dorsal position. The greater and lesser folds of the esophagus follow the course described by Fisher (1906, pp. 10-11) in Lottia. The lesser fold begins in the mid-ventral region of the pharynx, twists counter-clockwise as viewed from the rear, and finally reaches a dorsal position just before it ends. The greater folds, which begin dorsally, are twwisted so they come to lie ventrally at the posterior end of the esophagus. The twisting of the esophagus is the result of torsion in the veliger stage. Lateral pouches or sacculations are found between the greater and lesser folds. They become smaller and finally disappear as the esophagus curves to the right in the mid-visceral region (Figure 7). The paired buccal glands lie along or under the mid¬ esophagus. Their ducts, which overlie the head muscles and empty into the oral cavity on dorso-lateral folds, are white and prominent. The numerous posterior salivary, or esophageal glands lie on either side of the pharynx dilation and esophagus. The are conspicuous, small, finger-like processes that Catherine Gene Walker -5- extend out laterally and, in some species, ventrally around the esophagus. These glands open into tiny pockets, which in turn are divisions of the lateral sacculations of the esophageal wall. Like the esophageal folds, the glands and pharynx dilation appear twisted as a result of torsion. The esophagus passes into the proventriculus or forechamber of the stomach, which opens into the stomach by way of a contractable aperture encircled by numerous small folds. The stomach forms a wide loop, encircling the digestive gland. In sexually mature individuals the gonad is sometimes visible laterally beyond the stomach margin. The digestive gland occupies the central region of the visceral hump, slightly overlapping the inner margin of the stomach and almost completely covering the right lateral pootions. A single digestive gland janction with the csophaqus. duct enters the stomach near its prorimalend. The volume and color of the gland vary somewhat with species and with the size of the animal. The hind gut makes a series of four loops before it empties into the rectum. (Figure 2). Loops 1 and 4 circle clockwise, while Loops 2 and 3 circle counter¬ clockwise. The rectum begins where Loop 4 crosses the anterior portion of the stomach. Constriction of the feces into linked pellets occurs in the latter portion 4/3 Catherine Gene Walker -6 of Loop 4 and the rectum. The pellets are readily broken up, making them easily washed out of the nuchal cavity when the animal is splashed. Often if the animal has not been exposed to water for some time, fecal pellets may fill both sides of the mantle cavity. In the comparative study of the gut in species of Acmaea, differences between species were observed in the length and placement of the radula sac in the body, the jaw, the salivary glands, and the gut loops. LENGTH AND PLACEMENT OF THE RADULA SAC The radula sac of all species except Acmaea scutum extends only to the mid-visceral region (Figures3 and 4). The radula sac of A. scutum passes farther to the left and extends to the posterior visceral region before passing anteriorly again (Figure 4). The radula sac of A. scutum is almost twice the length of the shell, while in the other species it rarely exceeds 1.5 times the shell length (Table 1). THE JAW For species comparison, the jaw was removed, the muscle scraped away from the lower side, and the jaw then examined microscopically. When jaws were mounted vater in fluid on slides, the cover slips were slightly prop- then ped up to avoid breaking the jawsor bending severely. Approximately 10 jaws, taken from animals ranging from small to large, were examined for each species (Figure 5). Catherine Gene Walker -7- The jaw remains the same size, relative to the size of the animal, as growth occurs. With age, however, the anterior band gets harder and darker. The median ridges of the band become more prom-inent, possible as a result of wear caused by scraping of the radula, In Figure 5 the jaws of large adult specimens of the species studied are arranged according to an increase in the irregularity of the anterior band. Acmaea asmi, A. limatula, and A. scutum show little or no marking on the band, and the anterior margin is smooth and unridged. A. scabra, A. digitalis, and A. paradigitialis have small ridges in the medial region of the band, causing the margin to be slightly uneven. In A. insessa, A. pelta, and Lottia gigantia the anterior band bears a conspicuous median tooth with smaller ridges running along it. This characteristic is most apparent in A. pelta, which also has a darker anterior band. The undulating anterior border of the anterior band in the jaw of Lottia is rather distinctive. Species differences in shape of the lateral wings of the jaw are less apparent, though? variations occur in wing shape and in the relative size of the anterior and posterior wings. ANTERIOR AND POSTERIOR SALIVARY GLANDS The anterior and posterior salivary glands show some variation in placement and size in the species examined. In Acmaea digitalis, A. limatula, and A. scutum Catherine Gene Walker -8- the two buccal or anterior salivary glands extend along the esophagus, their posterior ends lying on the esopha¬ to the gus, rear of the posterior salivary glands (Figure 6). The ducts of the buccal glands of A. pelta, A. scabra, and A. asmi are shorter and wind back and forth several times across the head muscles (Figure 7). In these species the distal extremities of the buccal glands terminate near the posterior margin of the buccal mass on either side. In Acmaea digitalis, A. scabra, and A. limatula the very numerous posterior salivary glands extend straight out from the pharynx dilation and anterior esophagus (Figure 6). In A. pelta and A. scutum the pharynx dilation is wider than in the above species and the twist of the esophagus more apparent. (Figure 7). The glands here are slightly smaller, and curve ventrally around the foregut. In A. asmi the posterior salivary glands are very small, and light green in color, while in the other species they are white. A count of glands was attempted, but an exact number was difficult to obtain. The left side of the esophagus has about 65 glands, while the right side has a slightly smaller number, the number varying somehat with size of the animal and the species. LOOPING OF THE GUT The length and placement of the gut loops in six species of limpets is shown in Figure 8. Some variation Catherine Gene Walker -9 in the exact placement of the loops occurs within the individual species depending on the size of the animal and the degree of maturity of the gonad. However, there is a characteristic placement of the loops in each species. In Acmaea peltay A. scutum, and A. scabra Loop l circles farther posteriorly. Loop 2 is considerably smaller in A. digitalis, A. scabra, and A. asmi. In A. asmi the anterior portionsof Loops 1 and 2 extend farther anterior reltive to the stomach. In A. digitalis, and A. scabra Loop 3 does not lie as far to the left as in the other species. Dorsoventral thickness of the visceral mass is difficult to show in the drawings; in A. limatula and A. scutum, both rather flat species, the loops lie beside or just beneath one another. In the other species with taller shells the loops pass back and forth from ventral to dorsal regions as they twist through the digestive gland. The digestive tracts in A.pelta, A. limatula, and A. scutum are somewhat greater in diameter than those of the other species; this is a consistent difference, which appears related to diet but shows up whether the gut is full or empty. The stomach of A. pelta often contains sizable fragments of larger algae, and this species has been shown by Craig (1966) to feed mainly on larger red, brown, and green algae. The gut in A. limatula, which feeds mainly on flat encrusting red algae (Eaton, 1966) contains smaller fragments. The gut contents of A. scutum ob¬ Catherine Gene Walker -19- served in the present study resembled those found in A. limatula. In contrast A. digitalis, A. scabra, and A. asmi are known to eat microscopic green and blue¬ green algae and diatoms (Castenholz,1961; Fritchman, 1961; Haven, 1965). The gut contents of these three species consists at very finely divided material. Looping of the gut is less complex in the smallest limpets. In order to determine at what stages the small limpets develop the adult pattern of intestinal coils, small limpets, 0.5 - 5.0 mm in shell length, were examined. Animals 3 to 5 mm long could be identified to species. They were preserved and dissected ;in the usual way. Results showed that the digestive tract was present in essentially the adult condition. Still smaller individuals (1-2 mm in shell length) probably represent¬ ing Acmaea scabra and A. digitalis were collected in a small splash pool in the high intertidal zone, which contained only large A. scabra and A. digitalis. Minute specimens (0.5 - 2.0 mm in shell length) of these species and probably A. pelta as well, were also collected.in the crevices among Mytilus californianus Conrad 1837, Tetraclita squamosa Darwin, 1854, and Mitella polymerus (Sowerby, 1833). These tiny limpets were killed and preserved in 70% alcohol, dehydrated in an alcohol series (Ehe shells being removed in 95% alcohol), and cleared in cedar wood oil. The foot and Catherine Gene Walker -11 mantle was removed by dissection, and the animals mounted on slides in cedar wood oil. The limpets had been feeding, and the digestive tract was dark and clearly distinguishable through the remaining more transparenfk tissues. The results of these studies are shown in Figures 9 - 11. Limpets 0,5 - 1.5 mm in shell length have a relatively short stomach and intestine, and intestinal Loops 1, 2, and 3 are not evident, though there are twists in the gut. Loop 4 and the rectum are :in the normal position. Limpets 1.5 - 2.5 mm in length have Loops 1, 2, and 3 but these are not fully developed (Figure 12). When the shell reaches 2.5 - 3.5 mm in length the digestive tract is almost the same as that in the lrger animals (Figure 13). Placement of the inner Loops 1, 2, and 3 may be slightly out of regular orientation, but this displacement is random. The radula sac is easily seen in its normal posifion. COELOMIC SYSTEM The coelomic system consists of four interconnected cavities in the visceral mass: the right and left kidneys, the pericardium, and the gonad. The points of inter- connection lie anteriorly on the right side of the visc¬ eral mass, just behind the muscular wall of the nuchal cavity (Figure?12). The right kidney, easily identified by its dark green to brown color, encircles the whole visceral mass, Catherine Gene Walker -12 with its distal end just posterior to the pericardium. Internally its surface is quite irregular. The large right renal-genital pore has thick muscular lips and is just to the right of the anal opening. The pore on Acmaea scabra is much larger than that in the other species, and even exceeds the size of the anus. The left kidney is much smaller and difficult to distinguish from the muscular wall separating the pallial and visceral cavities. It is situated just back of the nuchal cavity, between the midline and the anus, and overlies part of the rectum (Figure 12). A. digitalis and A. scabra both have relatively large left kidneys. The left renal pore is a narrow slit opening on the anterior surface of the middle of the kidney (Figure 13). Only in A. digitalis are there very small bulbous lips on each side of the slit. The pericardium occupies the space above the gonad reoy in the left dorsal anterior a of the visceral mass. From it the reno-pericardial canal (Figure 13) crosses the body from left to right and, as in Lottia gigantea (see Fisher, 1904), divides just under the left kidney, one canal going to the left kidney and one under the rectum to the right kidney. The gonad is situated in the hollow above the foot beneath the digestive system. During the breeding season when it is greatly enlarged it extends up around the qut. The duct is merely an extension of the thin 3. 4. Catherine Gene Walker -13 epithelium that surrounds the gonad (Figure 13). It arises on the antero-dorsal wall of the gonad in the midline and extends to the right to open into the right kidney. Gametes pass from the gonad to the right kidney, and leave the kidney through the right renal-genital pore. SUMMARY Placement and extent of the radular sac, the form 1. of the jaw, the arrangement of the salivary glands, the looping of the gut, and the interconnections of coelomic derivatives are described and illustrated for Acmaea: A. asmi, A. digitalis, A. limatula, A. pelta, A. scabra, A. scutum, A. insessa, A. para- digitalis. Some notes on Lottia gigantea are included. The radula of A. scutum is almost twice the length of its shell, while in the other species the radula ranges from 1 - 1.5 times the length of the shell. Limpet jaws vary primarily in the shape and character of the hard anterior band. The jaws of K. pelta, A. insessa, and Lottia gigantea bear an anterior median tooth. The limpets studied can be separated into two groups on the basis of the salivary glands. A. digitalis, A. limatula, and A. scutum.have buccal glands that extend down the esophagus posterior to the esophgeal glands, while in the other species they terminate near the posterior margin of the buccal mass. 45 6. Catherine Gene Walker -14- In A. pelta and A. scutum the posterior salivary glands are slightly smaller, and curve ventrally around the foregut, while in the other species the glands etend straight out from the pharynx and esophagus. The pattern of gut loops showa minor but consistent differences between Acmaea species. Development of the adult pattern of loops is essentially com¬ pleted before animals reach a shell length of Amm. The gut tends to be thicker and heavier in species feeding on larger erect or encrusting algae (A. pelta, A. limatula, A. scutum) than in species feeding on films of microscopic algae (A. digitalis, A. scabra A. asmi). The coelomic cavities and ducts are described and diagramed. Both right and left kidneys of A. digitalis and A. scabra are relatively larger than those of other Acmaea species. Aneexcellent paper by Righi (1966), published after the present study was completed, describes the internal anatomy of the Brazilian species of Acmaea and provides illustrations permitting comparisons to be made between Brazilian and Californian species. Righi also finds that the shape of the jaw and the looping of the gut provide means of distinguishing Acmaea species. 48 Catherine Gene Walker -15- ACKNOWLEDGMENTS My sincere thanks go to Dr. Donald Abbott for his encouragement and guidance throughout this project. Thanks also are extended to the staff of Hopkins Marine Station. This work was made possible by Grant G4806 from the Undergraduate Research Participation Program of the National Science Foundation. 42 Catherine Gene Walker -16- LITERATURE CITED Castenholz, Richard W. 1961. The effect of grazing on marine littoral diatom populations. Ecology 42 (4): 783-794, 6 figs. Craig, Peter C. 1966. The activity pattern and food habits of the limpet, Acmaea pelta. The Veliger Eaton, Charles M. 1966. The Activity and food of the file limpet, Acmaea limatula. The Veliger Fisher, Walter K. 1904. The anatomy of Lottia gigantea. Zool. Jahrb., Anat. 20 (1): 1-66, 13 figs., pls. 1-4. Fretter, Vera, and Alastair Graham 1962. British Prosobranch Molluscs. The Ray Society, London. 1-xvi + 755 pp., illus. Fritchman, Harry K. 1960. Acmaea paradigitalis sp. nov. (Acmaeidae, Gastropoda). The Veliger 2 (3): 53-57, pls. 9-12. Fritchman, Harry K. 1961. A study of the reproductive cycle in the California Acmaeidae (Gastropoda). The Veliger: Part I, 3 (3): 57-63, pl. 10. Part II, 3 (4): pls. (5-17. 95-101, Part III, 4 (1): 41-47, pls. 9-14. 123 Catherine Gene Walker -17- Haven, Stoner B. 1965. Field experiments on competition between two ecologically similar intertidal gastropods. Bull. Ecol. Soc. Amer. 46 (4): 160. G. Righi, 1966. On the Brazilian species in the Acmaea subrugosa complex (Gastropoda: Prosobranchia: Patellacea). Malacologia 4 (2): 269-295, 27 figs. Runham, N. W. 1963. A study of the replacement mechanism of the pulmonate radula. Quart. Jour. Micros. Sci. 104 (2): 271-277, 3 figs. Test, Avery R. (Grant) 1945. Ecology of California Acmaea. Ecology 26 (4): 395-405. Test, Avery R. (Grant) 1946. Speciation in limpets of the genus Acmaea. Contrib. Lab. Vert. Zool., Univ. Michigan 31: 1-24 42 SPECIES A. limatula A. insessa A. digitalis A. paradigitalis A. scabra A. asmi A. pelta Lottia gigantea scutum NUMBER OF SPECIMENS EXAMINED 10 10 10 10 10 10 10 10 14 -18 TABLE 1 SIZE RANGE SHELL LENGTH cm. 2.0 - 3.5 1.5 - 2.0 1.5 - 2.5 1.0 - 2.0 1.0 - 2.5 0.5 - 2.5 1.3 - 3.5 3.0 - 5.6 1.5 - 4.0 Catherine Gene Walker RADULA LENGTH RATIOMOF SHELL LENGTH RANGE MEAN o.90 - 1.29 1.06 0.97 - 1.33 1.07 0.90 - 1.25 1.13 1.00 - 1.40 1.19 1.00 - 1.40 1.20 1.06 - 1.37 1.23 1.06 - 1.50 1.24 1.07 - 1.58 1.29 1.71 - 2.00 1.90 2 Catherine Gene Walker -19- CAPTIONS Figure 1: Generalized dorsal view of an Acmaea with shell and mantle removed: AN - anus, GPT - cephalic tentacles, CT - ctenidium, DG - digestive gland, E - eyespot, ET - foot, GN - gonad, RCT - rectum, RS - radula sac, SM - shell muscle, ST - stomach. Figure 2: Generalized dorsal view of the digestive tract of an Acmaea spe with the dorsal body wall, kidneys, gonad, digestive gland, and pericardium removed: AN - anus, DGD - digestive gland duct, ES - esophagus, J - jaw, RRO - Proventriculus¬ RCT - Rectum, RS - radula sac. Figure 3: Dorsal view showing the placement of the radula sac in the visceral cavity of Acmaea limatula (shell length - 2.5cm.; ratio of radula lenghh: shell length - 1.11). Placement of the sac here is similar to that in A. pelta, A. scabra, A. digitalis, and A. asmi. ES - esophagus, RC - Radula caecum, RS - radula sac, ST - stomach. Figure 4: Dorsal view showing the placement of the radularsac in the visceral cavity of Acmaea scutum (shell length - 2.5cm; ratio of radula length: shell length 7 1.9). ES - esophagus, RC - radula caecum, RS - radula sac, ST - stomach. Figure 5: Dorsal viewsof the jaws of large adult limpets. The jaws are partially flattened out on slides and drawn with a camera lucida. Dark areas represent the most opaque areas. Ke Catherine Gene Walker -20- Figure 6: Dorsal view of the anterior end of Acmaea digitalis, showing the location of the anterior (buccal) and posterior salivary glands. Semi-diagrammatic, based on 15 adult individuals. BG - buccal gland, BGD - buccal gland duct, ES - espphagua, GN - gonad, J - jaw, PD - pharynx dilation, PSG - posterior salivary glands, RS - Radula sac, ST - stomach. Figure 7: Dorsal view of the anterior end of Acmaea pelta, showing the location of the anterior (buccal) and posterior salivary glands. Semi-diagrammatic, based on 15 adult individuals. BG - buccal gland, BGD - buccal gland duct, GF - Greater folds, J - jaw, LF - lesser folds, PD - pharynx dilation, PSG - posterior salivary glands, RS - radula sac, SAC - sacculations. Figure 8: Diagrams of the stomach, intestine, and rectum of adult individuals of six species of Acmaea,based on dissections of 25 - 30 animals of each species. Exact placement of the loops is subject to slight variation, within each species; the diagrams show the typical condition in large adults. RCT - rectum, ST - stomach. Figures99, 10, 11: Generalized diagrams of Acmaea digitalig and A. scabra, showing the development of the digestive tract in dorsal view. RCT - rectum, ST - stomach 42 Catherine Gene Walker -21- Figure 12. Generalized dorsal view of the coelomic system in Acmaea, with the dorsal body wall, digestive system, and posterior te wall of, nuchal cavity removed. AN - anus, GN - gonad, LK - left kidney, P - pericardium, RK - right kidney, RRP - right renal pore. Figure 13: Enlarged view of the right upper corner of Figure 12, showing the interconnection of the coelomic cavities. Kidneys and anus open into the nuchal cavityl. AN - anus, CT - ctenicium, GN - gonad, GND - gonad duct, LRP-left renal pore, PRCr pericardial renal canal, RK - right kidney, RRP - right renal pore. Permanent address: -22- FOOTNOTES 43 Catherine Gene Walker Cer¬ CT- GN- RCT- OG- 57. ua? 8S¬ Ber Lop3 PRo- Loel jguve . 1 . -85 K-1 -80 -89 —AN 18 -100P2 ++ e 0 es . . .. .. . . ka- . . ea iadeinihäini 5 . . 4 . e t ... . * ..... . . .. Wi . . . . . . . .. ... .... .. . . ... .... Eis apo . . . - . . * . . . . „ . . . . 1/4 . . (. N Fr. 2 88 0 859 7. . /. Vr . ( - -— ee, . 5--- 4 V W r NA R W eiein 9 .pet 6 p 00P. L 10003 p3 Loi. A.dig talis A.nalula AScabre A Sun r ig (--—- - Ae L - D P t . . r . Wii S i 1 . „ : * .. A — Be -ST oo1 i 1.5 -5m 8e1 +1003 L Fiue 11 2.5-.5m Lop2 p3 Lop Kg X10 7 .*.. R .. :.- . . . .* 5 2 &Z ---- . . ... 77 . ....