Introduction Orbinia johnsoni (Moore, 1909) inhabits, according to Smith and Carlton (1975) protected sandy beaches. This species is known from Marin County to La Jolla, California, in both low intertidal and subtidal locations. Hartman (1957) describes the worm as being 80-90 mm long, but the worms used in this study, after being relaxed in magnesium chloride and preserved in formaldehyde, were found to range from 161 to 225 mm, with a mean length of 190 mm. Though the descriptions of Moore (1909) and Hartman (1957) are full descriptions, they do not give any information on the biology and feeding behavior of the orbinids. This study proposes to contribute information on these two factors. Two questions are raised. First, what is the population density of the worms? Second, what is their feeding behavior? The latter concern is further divided into four categories. First, when do they eat? Second, how long does it take for food to travel through their gut? Third, what do they eat? Fourth, where do they find their food? Materials, Methods and Results The population density of Orbinia johnsoni from the Boatworks Beach (fig. 1) east of Mussel Pt., Pacific Grove, California, was measured by taking a transect between 3.5 ft and -1.0 ft. tide level. The worms were found between 3.1 ft. and -0.5 ft., seaward of a pop¬ ulation of Nerinides acuta (Spionidae). Only occasional Nephtys californiensis (Nephtyidae) occurred with Orbinia. The sand grain size on the beach is fairly homogeneous (+1-+20). The worms are uncovered 3 to 4 days out of a monthly tidal cycle. To collect the worms, a hollow metal cube with a volume of 15,625 cubic cm, enclosed on four sides and open at the top and bottom, was pounded into the sand at intervals of 2 m throughout the length of the transect. The cube was pushed into the sand until the upper limits were flush with the surface of the sand. All the sand inside the cube was then dug out and placed on a wire screen with holes larger than the sand but significantly smaller than any organisms living in the sand. Seawater was poured upon the screen to filter the sand through it. After all the san was drained away, all the orbiniids living in the sand remained scattered upon the screen. They were then counted and recorded (fig. 2). Nine samples were taken. The range was 7-94 worms per sample. The mean density was 27 orbiniids per 15,625 cc. When the worms eat was investigated by collecting worms at various times during a 36-hr period, coinciding with a series of low tides. Comparing volumes of fecal material from different times of day with respect to tidal levels could disclose if there was periodicity in their eating habits that conformed to the rise and fall of the tide. Worms were collected at 6-hr intervals and at every low tide for a 36 hr period. Ten worms were obtained during each collection. Each worm was brought to the laboratory within one-half hour after collection, cleaned thoroughly of sand and mucous, and placed in a petri dish partially filled with sea water. The worms seemed to defecate all of their gut contents in 5-6 hours. The petri dishes were kept on a sea table with running water for as much as 36 hours. The 36-hr collection yielded ninety worms. The last thirty worms, however, were placed in syracuse dishes rather than petri dishes. All of these worms died before they had defecated completely; death was attributed to the small size of the dishes. Only the first 60 fecal pellets (obtained from the first 24 hrs of collection), consequently, were used for determining fullness of the guts. After 36 hours the fecal pellets were removed from the petri dishes using a pipette. The volume of each fecal pellet was measured by placing it in a graduated centrifuge tube filled with sea water. The feces, mostly sand, would always quickly settle to the bottom. The volume of each worm was measured by the amount of water it displaced in the tube. A ratio was attained for each worm's defecation by dividing the volume of the fecal pellet by the volume of the worm after defecation. The mean for each of the six sets of ten ratios was attained to arrive at six separate data points. The fecal pellets were then mounted on slides and preserved for later examination. A comparison of attained volume feces/volume of worm ratios to tidal cycles within the 24-hr period shows a parallel relation between high ratios and high tides, in addition to the close relation between low ratios and low tides (fig. 3). All of the data points, however, have extremely wide ranges. Some of the ranges vary from 0.0 to 1.0. The standard deviation for each of the data points is extremely large, making the means statistically insignificant. A follow-up study was conducted in search of further evidence that might support the finding that orbiniids have full guts during high tides and empty guts during low tides. Worms were collected every three hours for a 24-hr period. Eight sets of ten worms, the product of the collection, were placed individually in separate petri dishes partially filled with sea water which were, in turn, kept on a sea table. Again, this procedure was completed for each set of ten worms within one-half hour after collecting them from the field. They were left to defecate for 36 hours. Ratios of volume of feces/ volume of worm were acquired by the same methods applied in the preceding experiment. The means for each of the 8 sets of ten ratios was obtained to arrive at 8 separate data points. The 80 fecal pellets were then mounted on slides and preserved for later use. The mean defecation over time does not appear to conform to tide levels (fig. 4). Once again, ranges and standard deviations are very large. Comparing all 140 of the defecation ratios to tide levels in a scatter diagram, where each fecal pellet's relative volume is represented individually sheds more light on the extent to which feeding behavior could be a function of tide (fig. 5). Here one notes low ratios clustered about low tide levels and high ratios clustered about high tide levels. Furthermore, primarily low ratios occur during outgoing tides and mainly high ratios are found in incoming tides. Determining how long it took for food to go through the worm's gut was done by placing worms for one hour in petri dishes that were partially filled with methylene blue stained sand. Then the worms were cleaned off and moved to clean petri dishes filled partially with sea water. The dishes wereplaced on a sea table and spot checked every fifteen minutes for defecation of blue sand. Fig. 6 summarizes the results of the four worms that did defecate the blue sand. Discerning what the orbiniids ate was done by analyzing the preserved fecal pellets retained from the earlier experiments. 170 slides were analyzed. These were compared to the composition of sand samples from the orbiniids' habitat (fig. 7). The worms were found to eat about 932 sand, 42 organic detritus, 12 diatoms and less than 1 percent of both blue green algae and bacteria. Not all the animals, however, had extremely high percentages of sand in their fecal pellets. Some had as low as 5 percent sand and as high as 95 percent organic matter in the feces. This large variation justifies an analysis of the percentages of organic matter in the fecal pellets at varying tide levels. Such an investigation might offer an explanation for the large percentages of organic detritus in some feces. Fig. 8 shows that 15 of the 16 fecal pellets with organic matter constituting more than 20 percent of the feces were collected during tide levels of 2 ft or lower. Furthermore, the majority of these worms were collected during an outgoing tide. An examination of the size of the fecal pellets that have high percentages of organic matter reveals that they are relatively large in size (fig. 9). A breakdown of the feces by percentage of organic composition shows that unidentifiable organic detritus accounts for most of the increase in amount of organic matter (fig. 10). To find out where in a column of sand the worms get their food 3 narrow aquaria, each 40 cm long, 40 cm deep, and 1.5 mm wide, were filled with sand. One orbiniid was put in each aquarium with constantly circulating sea water. The aquaria were placed in a darkroom lit by a dim red light. Observations were made on the movements of the thorax in two overlapping twelve hour shifts (fig. 11), in order to determine vertical movement and hence food-getting behavior. Discussion The population density of the orbiniids is most highly concentrated at 0.0 ft tidal level. Here they are found 27 per 15,625 cubic cm. The worms are not in uniform concentrations but are clustered. Food apparently becomes available on incoming high tides since the guts are full at this time, even though there is a lack of statistical significance to prove this (fig. 7). It is conceivable that the incoming tide triggers their feeding response since high tide finds them with their guts the fullets. It takes 3-3.5 hrs for food to pass through the gut which is ample time for feeding during an incoming to high tide, defecate during outgoing to low tide and be ready to feed again for the next high tide. Though the average worm appears to ingest sand either preferentially or concurrently with other food (fig. 10), some worms showed upward of 512 of their fecal contents (fig. 10) as being derived from organic material. A possible explanation of this might be in being near a pocket of decaying seaweed which commonly occur on this beach. During outgoing tides the receding wave action has a tendency to leave rows of wrack on the beach. This wrack could seep into the sand when the tide is out and eventually provide a food source for the worms. The worms are probably bottom feeders. In the field they were never observed to be closer than 10 cm below the surface of the sand. In aquaria their tendency was to go to the bottom and stay there. Summary Orbinia johnsoni were studied from a small sandy beach near Mussel Pt., Pacific Grove, California, where they are most abundant at low tide levels in fine sand. They were found to have a mean length of 190 mm. The orbiniids were found with their guts the fullest during incoming to high tides. It is plausible that this is when they are feeding. It takes about 3 to 3.5 hrs for food to travel through the length of the gut. The orbiniids eat 93 percent sand and seven percent organic detritus. Special note should be taken that some food selectivity appears to be involved and that high percentages of organic matter in the feces are found in worms collected during low, outgoing tides. Evidence suggests that the worms are bottom feeders, not coming to the 0 surface to feed. ACKNOWLEDGEMEN I wish to acknowledge Dr. Isabella and Dr. Donald P. Abbott for their time, patience and endless energy. References Hartman, 0. 1957. Orbiniidae, Apistobranchidae, Paraonidae. Allan Hancock Pacific Expeditions 15: 257. Moore, J.P. 1909. Polychaetous annelids from Monterey Bay and San Diego, California. Proc. Acad. Nat. Sci. Phila. 161: 260-262. Smith, R. and J. Carlton (eds.). 1975. Light's Manual, Intertidal Invertebrates of the Central California Coast. 3rd ed. Univ. of Calif. Press. Berkeley. 716 pp. FIGURE CAPTIONS Figure 1: Map of a portion of Boatworks Beach, showing location of Orbinia johnsoni. Figure 2: Population density per 15,625 cm’. Mean density: 27; range: 7 to 94; sample size: 9. The population is most highly concentrated at around O foot tidal level. Figure 3: Mean defecation over time compared to tide levels.: Solid line: tide level; dotted line: mean defecation. Figure 4: Mean defecation over time compared to tide levels. Solid line: tide level; dotted line: mean defecation. Figure 5: Defecation ratios compared to tide levels. V: the worm was collected during an outgoing tide;A: the worm was collected during an incoming tide. Figure 6: Time for food to go through gut. Figure 7: Detritus: feces compared to sand samples. A: sand; B: organic detritus; C: diatoms; D: blue-green algae; E: bacteria. Figure 8: Organic detritus (2) in feces at varying tide levels. V: the worm was collected during an outgoing tide; A: the worm was collected during an incoming tide. Figure 9: Percentage of organic matter in feces compared to fecal volume. Each dot represents one fecal pellet. Figure 10: Breakdown of feces by percentage of organic composition. A: sand; B: organic detritus; C: diatoms; D: blue-green algae; E: bacteria. Figure 11: Positions of thorax in aquaria. Solid line: mean depth of thorax along vertical axis, representing depth of sand; dotted line: mean distance moved each hour. 0 - sand level. FI GURE 1 LOWER LIMIT Vot — —0.5 1t UPPER LIMIT O O ROCK TRANSECT SCALE L tif L o N E - S — RATIO: VOL FECES/VOL WORM — 13431 3011 m 8 RATIO: VOL FECES/VOL WORM 1A31 3011 58 5 S I RATIO: VOL FECES, VOL WORN 55 WORMS — - m DETRITUS (%) IN FECES EACH SAMPLES SAND (%) 03 N s. J 5 S5 S I SS ORGANIC DETRITUS (%) 55 5 8 8 8 VOLUME (ML) . . . . . 0.2 o 5 8 5 8 -- - - — —-— - —— — — — — —- — -- A- DEPTH IN SAND (CM)