4 EXCRETORY PRODUCTS OF PAGURUS SAMUELIS AND PAGURUS GRANOSDANUS (Malacostraca: Decapoda) by Peter W. Brown Hopkins Marine Station of Stanford University Pacific Grove, California INTRODUCTION Although the excretory products of many crustaceans have been analysed and compared, Pagurus have received little study. Preliminary work revealed a large amount of urea in the excretory products of hermit crabs. This find- ing was unexpected, since most other crustaceans excrete ammonia as the principal product of nitrogen metabolism. A more complete investigation was undertaken to confirm the preliminary study. METHODS Pagurus samuelis (Stimpson, 1859) and Pagurus grano- simanus (Stimpson, 1859) were collected from the area around Point Pinos, the southern and most western tip of Monterey Bay, California. These crabs were brought into the laboratory and placed in small (30 ml.) beakers containing 15 ml. of sea water. The sea water was treated with antibiotics to prevent the growth of bacteria and decomposition of the nitrogenous waste products. Terramycin (oxytetracycline; intravenous; Chas. Pfizer and Co., Inc., New York) and streptomycin sul- fate (Eli Lilly and Gompany, Indianapolis) were used. The beakers were covered with cheesecloth to prevent the escape of the crabs and maintained at 15° C. for 12-18 hours. At the end of this time the crabs were removed, and the sample of excretory water was frozen-ferolater analysisater. The samples to be analysed were removed from the freezer and centrifuged to remove fecal pellets. A 12.5 ml. volume of the sample was then treated bytpassage through a 5 ml. column of Dowex 50Wx 8, in the sodium form, to remove most of the divalent cations present in the sea water. These cations precipitate on Nesslerization. The column was then washed with distilled water to a final volume of 25 ml. Total nitrogen, ammonia, urea, allantoic acid, and allan- toin were measured as ammonia by Nesslerization with or without preliminary treatment. Allantoin was cõnverted toi allantoic acid bytbasicahydrolysisnandathe allantoic acid to urea by acid hydrolysis. Urease solution prepared from urease tablets (Matheson, Coleman, and Bell) was used to convert urea to ammonia. Distillation of the ammonia was finally resorted to because of various precipitates that formed upon Nesslerization. Urease, allantoin, and allantoic acid were found to precipitate to various degrees with the Nesslers Reagent. The distillate was collected in 1 ml. of MI. O.1 N HCl, diluted 1with distilled water. 3 ml. of Nesslers was added and after 30 minutes the color was read in a Klett- Sommerson photoelectric colorimeter, with a blue filter. This value was compared to a standard curve. Uric acid was determined by the method of Folini(1930). RESULTS The amounts of the various products of nitrogen ex- cretion are presented in Table 1. While ammonia was the principal compound excreted from all the crabs tested, substantial amounts of other nitrogenous wastes were found. The results suggest the mechanism for dealing with excess purines is through a total conversion to NH.. Most of the intermediates in the mmetabolic series for the degradation of purines are found to some extent in the excretory water of these animals. The amount of urea excreted showed great variation; it was an important product excreted by somgcrabs and was absent from the excretroy water of others. No marked difference in the excretory products of the two species was detected. TABLE 1 Excretory Products of Pagurus samuelis and Pagurus granosimanus ALLANTOT ALLANTOTN UREA NH, ACTD P. Samuelis 67.6* 74.4 10.5 42.0 149.0 120.0 27.0 103.8 101.2 P. granosimanus 22.8 2.0 178 13.4 12.0 217.2 44.6 137.4 34.2 188.4 xAll table values in ug N/ml. of undiluted excretory water. URIC ACTD 2.46 42 .86 .42 1.30 DISCUSSION Pagurus samuelis and P. granosimanus seem to follow the pattern set by other crustaceans, i.e. they are ammonotelic, the principal waste product of nitrogen metab- olism being ammonia. (See Waterman, and Prosser et al). The presence of large amounts of urea excreted by some of the crabs was a surprising result and is a deviation from the norm established by other members of the class. This fact suggests more extensive study to determine the extent of these large excretions of urea in other Pagurids of the same and different species. If this trend continues, espec- ially in species of the upper intertidal, it could indicate that a least one of the necessary metabolic adaptations for a more terrestrial way of life is possible in this group of crabs. SUMMARY While ammonia was found to be the principal nitrog- enous excretory product of Pagurus samuelis and P. granosimanus, substantial amounts of urea, allantoic acid, and allantoin were also detected. Small amounts of uric acid are also excreted. 2. Individual crabs showed considerable variation in the amounts of the various nitrogenous compounds excreted. 3. No difference in excretory products was observed be- tween the species tested. BIBLIOGRAPHY An improved method for the 1. Folin, O. determination of uric acid in blood. J. Biol. Chem. 86: 179- 187, 1930. 2. Prosser, Cl Ladd and Frank A. Brown Comparåtive Animal Physiology, 2nd. edition, W. B. Saunders Company, Phila. and London. pp. 135-152. 3. Waterman, Talbot H. The Physiology of Crustacea, volume 1. Academic Press, New York and London. pp. 341-366. C