page 2 Cynthia Lebsack INTRODUCTION The chiton Mopalia lignosa (Gould, 1848) is a common inhabitant of the inter-tidal zone on Californian shores. Although it usually retreats to sheltered regions during the day and at low tide, it must be able to cope with relatively wide variations in temperature and salinity, both on a seasonal basis and during a single tidal cycle. Temperature and salinity in the environment do not necessarily fluctuate together. In winters, for example, animals may experience relatively high salinities and low temperatures when submerged, and low salinities and low temperatures during rains at low tide. Studies of the combined effects of temperature and salinity on metabolic rates have been carried out on such marine organism as the crabs Hemigrapsus oregonensis Dana, 1851) and H. nudus (Dana, 1851) by Dehnel (1960), and the gill tissues of oysters and mussels by Van Winkle (1968). However, no studies of this sort have been made on chitons. I here report the effects of simultaneously imposed temperature and salinity variations on the respiratory rate of M. lignosa. All studies were carried out at the Hopkins Marine Station of Stanford University, Pacific Grove, California, during the period April-June, 1974. The chitons used in the experiments were collected at various points on the rocky shores of the Monterey Peninsula. Cynthia Lebsack page 3 MATERIALS AND METHODS Respiration rates were measured using Warburg- Barcroft manometers kept in a water bath held at a constant temperature. A wick of filter paper wetted with 0.2 ml of 10% KOH was placed in the side arm of the respiration chamber in order to absorb C0», and the vessels were agitated. The vessel constants were measured using a Gilmont-Warburg calibrator. Salinities were obtained by dissolving "Instant Ocean Synthetic Sea Salt" (Aquarium Systems, Inc.). in appropriate amounts of distilled water and were checked with a salinometer. Normal sea water from the environment (100%) was determined to be 33.91%. Animals taken from the field were held in running sea water at 13.510.5°0 for at least three days before use. I assumed the relation between respiratory rate (Vo2) and body weight (W) followed the equation; Voz-Kw0.74 where K is a constant. The value 0.74 is the generalized value for animals given by Prosser (1973) and is close to the average value (0.73) found by Kincannon (1974) for the chiton Tonicella Lineata (Wood, 1815). Using the above equation, the measured respiratory rates were corrected to that of a hypothetical ten gram chiton. The weight of the animals varied from 5.5-18.5 grams. Sample size varied from three to six animals at each temperature salinity combination. Cynthia Lebsack page 4 Individual chitons were placed in separate Warburg vessels and covered with seventy ml of synthe tic sea water at the salinity to be tested. They equilibrated for one hour at a given temperature. Readingswere then taken hourly for six hours. The average number of ul of 0 consumed/gram/hour was taken as the respiratory rate. Each of the three temperatures within the environmental range encountered by M. lignosa (800, 13.500, and 19°0) was combined with four different salinities (90%,100%, 110%, and 120% of normal sea water) to yield separate experimental conditions. Kincannon (1974) has shown that there is not conspicuous diurnal rythm in regards to chiton respiration. thus experimental runs were made at various times during a day. RESULTS Respiration rates vs. temperature for the four salinities are shown in Figure 1. Their 010 values are listed in Table 1. Like most poikilotherms. chitons respire at an increased rate at higher temperatures: go as. gar ou 36.2 ul at 100% sea water and 190C ys. 14.27 ul at 100% sea water and 800. The 010 value calculated using the 13.5°0 and values for animals in 1906 and 100 % sea water is 2.0. However, using the 80C and 13.50C values. a much higher 910 of 2.7 is obtained for the same animals. page 5 Cynthia Lebsa Respiration as a function of salinity for each experimental temperature is plotted in Figure 2. For each temperature, animals in 100% SW respired at a significantly higher rate than at any other salinity. DISCUSSION Most marine molluscs are osmoconformers with varying degrees of stenohalinity (Prosser, 1973). Kinne (1971), states that although not much is known on how salinity affects invertebrate metabolism, there are some general trends in the responses of species. One of these trends is that animals which are stenohaline decrease respiration at both the higher and lower salinites. This reduction in respiratory rate may be mediated through any of the following routes: (1) increasing or decreasing the body water or salt content; (2) changing of internal ion ratios; (3) hormonal, neuromuscular, and enzymatic interference and (4) behavioral changes (Kinne, 1971). In addition there is probably a reduction in the concentration of cellular metabolites in animals placed in hypotonic solutions, and a reduction of oxygen tension in those exposed to hypertonic media, both of which could lower respiratory rate. page 6 Cynthia Lebsack The least change in respiration due to osmotic stress was found, in both hyper and hypoosmotic solutions, at 8°0. Perhaps at this temperature the metabolism is nearly minimal for maintenance, and the chiton actively controls against further lowering. In 100% sea water the Qjobetween 8°0 and 13.5°0 is higher than that between 13.5°0 and 1900; this agrees with many observations on other animals that Q0 decreases with increasing temperature (Prosser, 1973). The Qio's increase from the lowest salinity to the highest for the colder temperature interval but remain quite constant at the higher temperature interval (Table I). Work should now be done to elucidate the physiological processes responsible for this low temperature change of 9jo with salinity. SUMMARY 1. Mopalia lignosa respire at a higher rate as the temperature increases. 2. Deviation in salinity from normal sea water causes a decrease in respiration rate. 3. At low, but not higher; temperatures, the 910 increases with salinity. Cynthia Lebsack page 7 Acknowledgments I would like to extend a sincere thanks to Christopher Harrold for his instruction on use of the Warburg, to Dr. Robin Burnett for his patience and help with statistics, and to Charles Baxter for his advice. aide, and encouragement on this project. To the staff and students of Hopkins Marine Station thank you for making this academic pursuit such an enjoyable one. Temp & Salinity Effects on Respiration, Mopalia lignosa page8 Lesback LITERATURE CITED Dehnel, Paul A. 1960. Effects of temperature and salinity on the oxygen consumption of two intertidal crabs. Biol. Bull. 118(2): 215-49; 215-49; 10 figs. Kincannon, Elizabeth A. 1975. The relations between body weight and habitat temperature and the respiratory rate of Tonicella lineata (Wood, 1815) (Mollusca-Polyplacophora) Kinne, otto 1971. Salinity: animals—-invertebrates. Pages 821-995 in Otto Kinne, ed. Marine Ecology. Vol. I, Part 2. New York, N.Y. (Wiley-Interscience) Prosser, Clifford Ladd, ed. 1973. Comparative animal physiology. xxii + 966 + xly pp.; illus. Philadelphia, Penn. (W. B. Saunders Co.) Van Winkle, Webster, Jr. 1968. The effects of season, temperature and salinity on the oxygen consumption of bivalve gill tissue. Comp. Biochem. Physiol. 26(1): 69-80; 5 figs. (28 December 1967) e Lebsack TABLE 1 values for the intervals 8°-13.5° C and for 13.5 C, for the four test salinities. page 9 o-19° page 10 Cynthia Lebsack FIGURE CAPTIONS Figure 1 - Mean oxygen consumption vs. temperature at salinities of 90%, 100%, 110%, and 120% sea water. Figure 2 - Mean oxygen consumption and standard errors vs. salinity at 8° C, 13.5° C, and 19° C. Letters next to individual data points indicate significant differences, using student's C-test, from 100% S.W. values obtained at the same temperature. =)p K.025; Cp.05. A2p.005; c Cynthia Lebsack TABLE 1 - ( Values TEMPERATURE INTERVAL 8°c - 13.5° c Seawater 1.5 90% 2.7 100% 110% 3.0 3.4 120% 13.5° c - 19° c 2.1 2.0 2.0 3.0 S 8 3 8 8 8 — 8