Kirsten Mattern Abstract The feeding behavior and prey selection of the deep-water calanoid copepod Gaussia princeps was observed under three different light wavelengths and four different food conditions. Both the free-swimming and tethered Gaussia showed no significant preference for any food type, light wavelength, or combination of food and light. Two possible foraging strategies and prey types of Gaussia are suggested. Comparison of the observed foraging strategies of the Gaussia to the known foraging tactics of the shallow-water calanoid copepods of similar structure show that Gaussia may be a suspension feeding herbivore or a cruise and sink feeding omnivore. This conclusion was made on te basis that the time copepods spend beating their mouthparts is a measure of foraging strategy. Kirsten Mattern Introduction Gaussia princeps is a calanoid copepod found in the mesopelagic zone. They have been found between 300 and 1000 meters and are thought to be vertical migrators (Bob North, pers. comm.). Both the adult male and female copepods are 9.0-12.0 mm in length. The bioluminescent capabilities of these crustaceans have been extensively studied, but much of their biology is unknown (Barnes and Case, 1972; Latz, Bowlby and Case, 1990; Bowlby and Case, 1991). The diet and foraging behavior of Gaussia has not been described, but the shallow-water copepod feeding strategies and food preferences have been widely characterized. The shallow-water calanoids are much smaller than Gaussia, but their similar features may help elucidate much about the deep-water copepods. These shallow-water copepods employ several different feeding strategies and select different prey types. The feeding strategies used by the shallow-water copepods are influenced by what type of prey they are seeking and the physical characteristics of their environment, such as light and water movement. By characterizing and comparing the feeding behavior of the Gaussia to the shallow-water calanoids, the foraging tactics and possible prey type of the Gaussia may be determined. The purpose of this research was to determine what type of prey Gaussia preferentially feeds on and what methods these deep-water copepods use to catch their prey. Kirsten Mattern Materials and Methods To examine copepod activity with different food and light types, I observed sixteen Gaussia each in two tanks maintained at 5°0 with a current created by continuous water flowing in a vertical circular pattern. As these crustaceans are accustomed to low light intensities, even a total absence of light, black sheets of plastic surrounded the tank, the front panel removable by velcro strips. The copepods were gently swept around by this current, and the current was necessary for them to remain suspended. lused three light types and four food conditions. White light came from the normal room light, UV light from an overhead light set on top of the observation tank, and red light from a normal light covered with two layers of red celophane, also set on top of the tank. I used the white light for a contrast from the light intensity the Gaussia encounter in their natural habitat and to provide a comparison to the shallow-water copepod environment. The UV light is absorbed at depths above that at which Gaussia have been found, and has been observed to cause the luminous cells in the copepods to fluoresce as well (Bowlby, 1991). Red light is also absorbed at a depth above that at which the Gaussia live and so allows the copepods to be observed in an environment closest to their own as they may be insensitive to this wavelength of light. The three food types consisted of phytoplankton, brine shrimp nauplii and natural zooplankton. The phytoplankton (Dunaliella and Nannochloropsis) and nauplii were obtained from the Monterey Bay Aquarium and the zooplankton was obtained from 10 minute tows done at Hopkins Marine Station. The zooplankton consisted primarily of shallow-water copepods, radiolaria, diatoms, cladocera, and chaetognatha. I did not feed theGaussia in one tank more than one food type per day, and the tank was cleaned regularly. Kirsten Mattern To specifically observe which food type Gaussia prefers and to determine how they capture their microscopic prey, I superglued individual Gaussia to a fine glass rod and suspended it in a sealed glass flask through which Ilet water flow with the different food types. observed the Gaussia under a microscope with white light. The cold temperature was maintained by packing ice around the flask. The appendages of the Gaussia and the food types were clearly visible under these circumstances. To observe individuals under each food condition, 1 placed 12 Gaussia in 12 separate glass tubes 4 inches tall and 1 inch in diameter. Three tubes had no food and served as the control, three had phytoplankton, three had brine shrimp nauplii, and the last three had zooplankton. I maintained the tubes in a 500 water bath. The tubes were also kept in the dark when not being observed. When I studied these tubes, I took them out of their water bath and set them under the different light types for 10 minute intervals to see whether the Gaussia displayed different behavior patterns under the different food conditions. Äfter the first week, I cleaned the tubes and rotated the food types, excluding the control tubes. I repeated this process after the second week of observations so each set of copepods was exposed to each food type. defined two types of behavior most often seen during my initial observations. One, the dart, is an abrupt voluntary swimming movement that was not a result of touching or colliding with another individual. Second, the side is a behavior where the individual hangs upsidedown facing the current, either with its dorsal or ventral side facing the tank wall. My observations of this copepod activity included watching them for 30 minute intervals and recording how often each type of behavior occurred per minute. I broke these time intervals into three 10 minute periods, each time using a different light source. observed the Gaussia for these 30 minutes under four different food conditions: Kirsten Mattern no food, phytoplankton, brine shrimp nauplii and zooplankton. For each 10 minute interval I calculated the averages of darts and sides. I conducted between two and five replicates of each food and light combination. Data were analyzed by two-way analysis of variance with light source and food type as fixed factors. Results Under the microscope, the Gaussia displayed several different types of movement patterns. When I kept the water flowing at a steady rate or at a zero velocity, the tethered Gaussia rapidly moved its antennae to create a current flow past its maxillipeds in the posterior direction. This pattern could be seen when added a dense suspension of food to the water. IfI suddenly changed the rate of water flow, the copepod would either completely stop all movement for a moment then resume activity with a rapid appendage contraction which included pumping of the legs and contraction of the maxillipeds and antennules, or immediately perform the rapid appendage contraction before resuming the antennae moyement. This movement of the legs and contraction of the maxillipeds and antennules are how the Gaussia propel themselves through the water, producing the dart behavior. I also observed this behavior when large pieces of debris associated with the plankton food suspension became entangled with the copepod's maxillipeds or antennules. The copepod could also move just the antennule or maxilliped on which the piece was stuck. When the copepod contracted its maxillipeds, they moved toward its mouth. This movement is extremely fast and although the microscopic plankton and individual nauplii were visible, I could not tell whether the Gaussia had Kirsten Mattern captured a prey or what prey type it was. The tethered copepods showed no specific preference for either plankton, algae or nauplii. Any contraction of the maxillipeds was sporadic and did not occur to a greater extent in any one food suspension. found no significant differences of copepod activity between each food type or between each light type. There was no significant interaction between food and light (Tables 1 & 2). Gaussia showed the most activity under white light with no food (Fig. 1). The least amount of dart activity occurred with the brine shrimp nauplii, regardless of light condition. The greatest average of individuals on the side per minute occurred under red light and with phytoplankton (Fig. 2). The side activity under UV light peaked with zooplankton. Under white light, the average activity remained fairly constant, regardless of food condition. Again, none of these differences were significant. When I placed the Gaussia in the glass tubes, they immediately sank to the bottom and made no attempt to swim unless the tube was disturbed. After I added each food type, no dart or side activity occurred. The copepods lay on either their dorsal or ventral surface and continued with the normal rapid antenna movement. This sinking behavior also occurred when I turned off the current in the large tanks. With no current, there was no activity under any combination of food or light. Discussion The foraging behavior of the shallow-water calanoid copepods has been extensively characterized. Koehl and Strickler (1981) investigated the sieve feeding tactic of the copepods. They scan the water by beating their feeding appendages, producing a postero-lateral current. The assymetric swinging of the Kirsten Mattern maxillipeds orients the copepod to capture the section of water containing the algae. Only the water containing the algae is pushed through the bristles on the second maxillae. The food particles are transferred to the mouth via the first maxillae. The preferred prey is phytoplankton, and these copepods can scan the water they filter for certain food. Captured prey can be ingested or rejected. Greene (1988) characterized the same filtering tactic used by the suspension feeding calanoids. He also described the omnivorous and carnivorous calanoids that recognize prey and seize the prey raptorially with the second maxillae and maxillipeds then use their first maxillae to guide the prey to their mouths. Greene (1988) stressed the strong interaction between feeding and swimming because these behaviors utilize the same appendages. He characterized three foraging tactics: the suspension feeders who remain stationary, the cruising, raptorial predators, and the omnivorous calanoids who employ a 'cruise and sink behavior. Tiselius and Jonsson (1990) described three hydrodynamically based foraging strategies of shallow-water calanoids. Stationary feeding is a balance between gravity and upward thrust, cruising copepods encounter more prey and can approach these prey in a hydrodynamically quieter manner than the stationary feeders, and the ambush predators sink slowly and passively and are hydrodynamically quieter than both the stationary and cruising calanoids. The time the calanoids spend moving their mouthparts is a measure of their foraging strategy. The upward thrust created by this movement counteracts the sinking brought about by gravity so the stationary feeders exhibit continuous movement while the ambush predators should only use their mouthparts to capture the prey. Gaussia exhibit the continuous movement of the mouthparts, or antennae, both in the presence and absence of food. These calanoids also sank to the tank floor and to the bottom of the glass tubes in the absence of a current, vet continued the movement. When in the tank, the upside down position on the Kirsten Mattern sides of the tank was also accompanied by the antennae movement. This may indicate that the Gaussia are suspension feeders, a strategy employed by the shallow-water algal feeders. In their natural habitat, Gaussia may be sinking while moving their antennae then moving upwards by the darting behavior, such as the cruise and sink method Greene (1988) attributed to omnivourous copepods. Based on data from shallow-water copepods, (Tiselius and Jonsson, 1990) the ambush method may be eliminated as a strategy of the Gaussia because they continuously move their mouthparts. The Gaussia may then be either suspension feeders or cruising feeders, depending on the current velocity, and their prey preference could be either herbivorous or omnivorous. Buskey, et al. (1989) found that certain oceanic copepods found ca 80m showed negative phototaxis over a wide range of light intensities, that is, the copepods were found in sections farthest from the light. Such copepods would then migrate downwards during the day and towards the water surface at night. This differs from the coastal and estuarine vertically migrating copepods which stay towards the bottom but within the photosensitive zone during the day According to Buskey et al. (1989) the maximum photosensitivity displayed by the deep-water copepods corresponded to the light wavelengths most prevalent in their environment and to the wavelengths of most marine bioluminescence. The exact effect of light on vertical migration is not clear. It is clear, however, that light can be used to predict the position of copepods if their photosensitivity thresholds are known. Light affects where the copepod is during a 24 hour period, and prey types at certain depths affect what strategy the copepod uses in its particular environment to capture the prey. Gaussia showed no differences in activity under any wavelength, white red or UV. Because they are deep-water copepods, they may be insensitive to the red and UV light because these light wavelengths are absorbed at a depth Kirsten Mattern above that which Gaussia inhabits. These light wavelengths could imitate their natural environment. The white light may have had some effect as the least amount of side behavior consistently occured under this wavelength. If light is not the factor that regulates preferred depth, Buskey et al. (1989) suggested that pressure may control the vertical migration of the deep-water copepods. To understand the foraging tactics and prey selection of the Gaussia to a greater extent, these calanoid copepods should be viewed in an environment more like their natural surroundings. The layer they live in not only has different water movements compared to the shallow-water environment and artificial tank currents, but is also under greater pressure. In addition, identity and abundance of the possible prey organisms in Gaussia’s natural habitat is necessary to fully determine what they eat. Two possible methods of discovering possible prey of the Gaussia are analyzing the gut contents of freshly caught Gaussia, and performing a fecal analysis. Defining the feeding biology of the mesopelagic calanoid copepod Gaussia princeps will not only show what types of species inhabit this deep zone, but will also help to determine the physical characteristics of this zone and how these species interact with each other and with their environment. Acknowledgements Thank you to Bob North at the Monterey Bay Aquarium for supplying the Gaussia and equipment for my research. would also like to thank Professor Mark Denny for his help with my set-up and with background material, and Professor Stuart Thompson for the use of his microscope. would also especially like to thank Professor Jim Watanabe for his help with research ideas, background material, statistical analysis, and reviewing my paper. Kirsten Mattern Literature Cited Barnes, A.T. 1972. Bioluminescence in the mesopelagic copepodGaussia princeps. J. Exp. Mar. Biol. Ecol. amp; 53-71. Bowiby, M.R., and J.F. Case. 1991. Ultrastructure and Neuronal Control of Luminous Cells in the Copepod Gaussia princeps. Biological Bulletin. 180: 440. 446. Buskey, E.J., K.S. Baker, R.C. Smith and E. Swift. 1989. Photosensitivity of the oceanic copepods Pleuromamma gracilis and Pleuromamma xiphias and its relationship to light penetration and daytime depth distribution. Marine Ecology Progress Series. 55: 207-216. Davis, C.C. The Pelagic Copepoda of the northwestern Pacific Ocean. Seattle, University of Washington Press, 1949. pp. 50-51 Greene, C.H. 1988. Foraging tactics and prey-selection patterns of omnivorous and carnivorous calanoid copepods. Hydrobiologia. 167/168: 295-302. 1981. Copepod feeding currents: Food Koehl, M.A.R., and J.R. Strickler. capture at low Reynolds number. Limnol. Oceanogr. 2666): 1062-1073. Latz, M.I., M.R. Bowiby, and J.F. Case. 1990. Recovery and stimulation of copepod bioluminescence. J. Exp. Mar. Biol. Ecol. 136: 1-22. Tiselius, P., and P.R. Jonsson. 1990. Foraging behavior of six calanoid copepods: observations and hydrodynamic analysis. Marine Ecology Progress Series. 66: 23-33. Kirsten Mattern Table 1: ANOVA of logdart: Average number of darts per minute during a 10 minute period. Data were log-transformed prior to analysis (In (x+1)). E-BATIO SOURCE MEAN¬ SQUARE 2.138 0.114 Food 0.450 0.778 0.05 0.253 Light Food X Light 0.12 0.574 0.748 0.2 ERROR 34 Kirsten Mattern Table 2: ANOVA of side: Average number resting per minute during a 10 minute period. SOURCE MEAN¬ E-BATIO of SQUARE 0.987 0.411 Food 1.12 0.057 3.568 3.128 Light 0.489 Food X Light 926 1.056 — 34 ERROR 1.141 Kirsten Mattern Figure Legend Figure 1: The average number of darts min during a ten minute period per light wavelength and food type. Figure 2: The average number of sides min during a ten minute period per light wavelength and food type. Kirsten Mattern FIGURE 1: AVERAGE NUMBER OF DARTS PER MINUTE. 3- — RED LIGHT —---- UV LIGHT ------: WHITE LIGHT 0 + NONE NAUPLII PLANKTON ALGAE 0 FOOD Kirsten Mattern FIGURE 2: AVERAGE NUMBER OF SIDES PER MINUTE —----- UV LIGHT WHITE LGIT ooo NONE NAUPLII PLANKTON ALGAE 0 FOOD