Trace metal levels in sandy beach Polychaetes of Monterey Bay, California W. Allen Shotwell Hopkins Marine Station of Stanford University Introduction Studies of trace metal levels in marine organisms have shown that toxic metals such as cadmium, lead and mercury are concentrated by many marine organisms (reviews by Vinogradov, 1953; Goldberg, 1965). Except for the studies of Bryan and Hummerstone (1971) on tolerance levels in Nereis diversicolor (Müller), Cross et al. (1970) on manganese, iron and zinc in estuarine polychaetes, and Phelps (1966) on iron, zinc, scandium and samarium, no research has been initiated on trace elements in polychaetes. This study is an investigation of levels of nickel, copper, cadmium, lead, iron, zinc and manganese in six species of typical beach polychaetes of Monterey Bay. Specific sampling sites were chosen so that known point sources of pollution in south Monterey Bay might be included in the survey. Materials and methods Collection sites on seven beaches in south Monterey Bay (Fig. 1) in an area extending from Hopkins Marine Station to Holiday Inn were chosen for study. These included the boat la Trace metals in beach Polychaetes works beach next to Hopkins Marine Station, the Monterey marina beach between wharf number one and two, a beach 500 yds, east of wharf number two, the Del Monte Apts. beach, the U.S.N.P.G.S. beach, the beach at the Monterey sewage pipe and the last, 50 yds. southwest of Holiday Inn in Monterey. Polychaetes were collected by sifting sand through a 1/8 inch plastic screen until sufficient numbers of organisms of a given species were obtained. Sand samples were also collected from those specific levels where each species was taken for later analysis. Specific sites for species were taken from a polychaete distribution study of south Monterey Bay by Anderson et al. (1972). The six species studied were: Cirriformia spirabrancha Moore, 1904 Euzonus dillonensis Hartman, 1938 Euzonus mucronata Treadwell, 1914 Nephtys californiensis Hartman, 1938 Nerinides acuta Treadwell, 1914 Travisia gigas Hartman, 1938 The worms were washed in seawater for 24 to 48 hours to purge them of attached sand and to allow removal of sand from the digestive tracts. The organisms were then dried for 12 hours at 72°C. They were subsequently ground to a powder with mortar and pestle and sifted through .5 mm. nylon netting. One gram samples were weighed out into 30 ml. beakers. Ten ml. of 90% HNO3 was added and the sample left uncovered under a hood for one hour. Samples were then refluxed one hour and evaporated to a volume of 5 ml. Five ml. of H90, was added Trace metals in beach Polychaetes dropwise until bubbling stopped and the resulting solution was evaporated to a final volume of 5 ml. One ml. of con¬ centrated HCl was then added and the solution was diluted to 25 ml. with distilled water. Sand samples were digested in the same manner in order to compare levels in the sand en¬ vironment with those in the polychaetes studied. The digested samples were analyzed by atomic absorption spectrophotometry. All collections of the polychaetes were pooled and two one gram subsamples were taken from the pooled tissue samples. All of the polychaetes studied are substrate ingestors with the exception of Nephtys californiensis, which is a predator. Feeding type was determined, in the case of the substrate ingestors, by dissection of the gut and microscopic analysis. N. californiensis was observed feeding on nematodes and other polychaetes. Results The results are presented in Figures 2-10 and Table 1. The elements will be discussed individually. Nickel: The nickel concentrations in the sand remain relatively consistent from beach to beach in a range of 4 ppm. to 10 ppm. (Fig. 2). Nickel levels do not vary greatly with different polychaete species showing a range of 3 ppm. to 7 ppm. (Fig. 4). The one exception to this is the opheliid Travisia gigas. At two different locations the levels in this polychaete are much higher (27 ppm. and 55 ppm.) than those seen in the other species (Fig. 4). The ratio of Trace metals in beach Polychaetes polychaete nickel concentration to concentrations in the sand are consistently in the range of .330 - .784, except for the ratios of T. gigas to the sand which are 8.30 and 4.09 (Table 1). Copper: Copper levels in the sand showed little varia¬ bility from transect to transect, with a range of 1 ppm. - 6 ppm. The polychaetes showed a ratio to the sand of over one, ranging from 3.00 to 12.6 (Table 1). The species with the highest ratios are Nerinides acuta with 24.0, Nephtys califor- niensis with 23.0 and Euzonus mucronata with 93.0. N. acuta and E. mucronata also showed the highest overall copper con¬ centrations of any species with 63 ppm. and 300 ppm. respec¬ tively (Fig. 5). Manganese: The levels in the sand were all over 10 ppm. with the highest concentration being 33 ppm. Manganese levels in the polychaetes were consistent from species to species ranging from 4 ppm. to 15 ppm. None of the ratios show any significant concentration of manganese in the polychaetes. Iron: Sand levels of iron were consistent with higher levels occuring at the boat works beach, 1400 ppm. and the harbor beach, 1500 ppm. The iron levels in any given species of polychaete were intraspecifically consistent from beach to beach. E. dillonensis and E. mucronata show a range of 750 ppm. to 860 ppm. T. gigas with a range of 350 ppm. to 420 ppm. and C. spirabrancha from the harbor showed the highest levels of up to 1700 ppm.; N. acuta and C. spirabrancha showed a ratio of 1 to the sand levels, and all others were from .10 - .75. Trace metals in beach Polychaetes Zinc: Levels of zinc in the sand were inconsistent from transect to transect, but showed a range of 6 ppm. to 32 ppm. The levels in the worms showed intraspecific consistency similar to that found with the iron levels. E. dillonensis had a range of 100 ppm. to 160 ppm. T. gigas had a range of 30 ppm. - 60 ppm. C. spirabrancha had 70 ppm. at both sites. The highest levels occur in N. acuta and Nephtys californiensis with 250 ppm. and 240 ppm. respectively. The ratios of the worm levels to the sand levels were all greater than 1:1 ranging from 3.06 to 3.14. The highest ratios were in N. californiensis, E. dillonensis from the U.S.N.P.G.S. and N. acuta. Lead: Sand levels were only detectable on the harbor beach and at the boat works beach. All species showed con¬ sistent levels of lead from 8 ppm. to 11 ppm. with the ex¬ ception of E. dillonensis and E. mucronata which had levels of 21 ppm. and 57 ppm. respectively. Cadmium: Cadmium was not detectable in the sand at any of the sampling sites. All species analyzed contained cadmium with the highest level in E. dillonensis of 11 ppm. Discussion All of the metal levels in the sand found in this study are consistent with surface sand levels found in a study by Koski (1972), with iron, zinc and manganese occuring in the greatest concentrations. Trace metals in beach Polychaetes The Monterey marina is directly adjacent to the Del Monte beach transect and the Monterey Boat Works is above the tran¬ sect at Hopkins Marine Station. These are sources of environ¬ mental pollution. It is interesting to note that the highest concentrations of lead and iron were found on the sand from these beaches. The polychaetes can be separated into accumulators and non-accumulators with respect to each metal. T. gigas seems to be the only nickel accumulator out of the six species studied. The consistently high ratios of copper found in the worms seems to indicate copper may be used in all of the species studied, possibly as a metallo-enzyme. Vinogradov (1953) cites evidence for a copper metallo-enzyme in annelids. The accumulators of copper seem to be Nephtys californiensis, E. mucronata and Nerinides acuta. These species show a ratio to the sand of an order of magnitude greater than the other species. All species seem to be non-accumulators of manganese as indicated by the ratios in Table 1. Annelids use three iron containing pigments, haemoglobin, chlorocruorin and haemerythrin (Dales, 1963), which accounts for the intraspecific consistency in the iron levels from transect to transect. The C. spirabrancha from the harbor shows a greater ratio of iron to the sand than C. spirabrancha from the Boat Works beach. This may reflect the presence of sand particles rich in iron in the gut or accumulation by the Trace metals in beach Polychaetes worm. However, the relatively consistent levels of iron in the same species coupled with the use of iron suggests regu¬ lation of iron in the polychaetes independent of the levels in the sand. The zinc accumulators seem to be N. californiensis and Nerinides acuta. The six species studied contain higher con¬ centrations than the sand. Phelps (1966) suggests accumula¬ tion of zinc in nine species of benthic polychaetes. They also show consistently high ratios of tissue zinc levels to the levels in the sand. Bryan and Hummerstone (1971) and Cross et al. (1970) give evidence for regulation of zinc con¬ centrations in polychaetes independent of the sand levels. The data from this study seems to justify such an hypothesis. All species studied appear to accumulate lead and cadmium above the levels found in the sand. E. mucronata seems to be the largest accumulator of lead. It is interesting to note that E. mucronata is also the largest accumulator of copper. While strongly suggestive, the data presented is not sufficient to confirm accumulation of these metals in the polychaetes investigated. This is a result of the time limit of six weeks and the scarcity of two of the species. (N. cali¬ forniensis and T. gigas occur in a density of approximately 1 per 1/4 m2 (Anderson et al., 1972).) Further research should be initiated to confirm the suspected accumulation of these metals. Nickel and iron levels in annelids suggested by Bowen (1971) are consistent with the levels recorded in this study. Trace metals in beach Polychaetes Summary There is evidence to suggest accumulation of nickel in T. gigas. Copper is accumulated in Nephtys californiensis, E. mucronata and Nerinides acuta. Accumulation of zinc takes place in N. californiensis, E. dillonensis and N. acuta. Lead accumulation seems to take place in E. mucronata. This study shows that zinc and copper are in consistently high levels in the worms as opposed to the sand and this may sug¬ gest some use by the worms such as a metallo-enzyme. Acknowledgements I would like to thank Dr. John Martin for his assistance in interpreting the data and his technical advice. I thank Dr. Welton Lee and Dr. Donald P. Abbott for their help in writing and editing this paper. Many thanks go to Mr. Keith Skaug for his invaluable assistance in the laboratory. Literature Cited Anderson, D. et.al. 1972. Polychaete distribution and related physical parameters on sandy beaches of South Monterey Bay, Cali¬ fornia. On file Hopkins Marine Station of Stanford Uni- versity. Bowen, H.J.M. 1971. Trace Elements in Biochemistry. Academic Press, New York. pp. 70-71. Bryan, G.W. 1971. The effects of heavy metals (other than mercury) on marine and estuarine organisms. Proc. Roy. Soc. London B Biol. Sci., vol. 177, pp. 389-410. Bryan, G.W. and L.G. Hummerstone 1971. Adaptation of the polychaete Nereis diversicolor to estuarine sediments containing high concentrations of heavy metals. J. Mar. Biol. Ass. U.K., vol. 54, pp. 845- 863. Cross, F.A., Duke, T.W. and J.W. Willis 1970. Biogeochemistry of trace elements in a coastal plain estuary: distribution of manganese, iron and zinc in sediments, water and polychaetous worms. Chesapeake Sci., vol. 11, pp. 221-34. Dales, R. P. 1963. Annelids. Hutchinson & Co. LTD., London. pp. 200. Goldberg, E. D. 1965. Review of trace element concentrations in marine organisms. Puerto Rico Nucl. Center., pp. 535. Koski, R.A. 1972. Trace metal concentrations on sandy beaches. On file Hopkins Marine Station of Stanford University. Phelps, D.K. 1966. Partitioning of the stable elements Fe, Zn, Sc, and Sm within a benthic community, Anasco Bay, Puerto Rico. International Symposium on Radioecological Con¬ centration Processes, Stockholm., Symposium Publica¬ tions Division, Pergamon Press, New York., pp. 721-34. Vinogradov, A.P. 1953. The Elementary Chemical Composition of Marine Or¬ ganisms. Yale University Press, New Haven, Conn. (Sears Foundat. for Marine Res.).. pp.640. Table A Boat Wks. Beach Harbor Del Monte Beach Del Monte Apts. U.S.N.P.G.S. Sewage Pipe Holiday Inn Table B N. californiensis dillonensis dillonensis dillonensis gigas dillonensis mucronata gigas spirabrancha C. spirabrancha N. acuta Appendix Sand metal levels in ppm. Fe Mn Pb cd Zn 12.7 =593 0.00 2.60 10.4 8.30 0.00 5.20 1400 14.9 16.7 0.00 26.3 6.50 6.20 15.5 1500 23.2 14.2 0.00 10.0 3.10 1160 12.9 0.00 0.00 23.8 1115 6.40 0.00 0.00 6.60 2.00 16.6 10.0 2.00 1141 32.0 0.00 0.00 24.7 6.60 2.00 16.6 1115 6.40 0.00 0.00 10.0 2.00 23.8 1141 6.40 0.00 0.00 6.60 2.00 32.7 1165 12.9 0.00 0.00 4.30 1.00 12.5 1700 7.60 0.00 0.00 Polychaete metal levels in ppm. Fe Cu Zn cd 23.0 3.2 239 182 3.3 11.3 3.2 5.1 16.5 3.8 748 106 7.0 2.0 4.1 6.6 16.6 4.0 818 161 858 6.6 25.2 5.3 100 18.2 6.7 55.0 6.1 7.0 367 52.3 O.00 1.0 38 6.6 4.0 801 128 22.7 11.4 4.0 299 3.3 2.6 15.1 419 59.0 1.1 27.0 6.1 6.8 5.1 1719 7.9 19.4 10.9 1.0 29.5 10.9 3.3 770 6.8 2.0 2.8 5.6 63.0 4.0 665 252 9.0 10 Table 1. Ratio of trace metal levels in each species to the trace metal levels of the sand in which that species was found. G. Nephtys californiensis F. Euzonus dillonensis E. E. dillonensis D. E. dillonensis D. Travisia gigas C. T. gigas c. E. dillonensis C. E. mucronata B. Cirriformia spirabrancha A. C. spirabrancha A. Nerinides acuta TABLE 1 Ni Cu Mn Fe .767 23.0 .256 .107 72 8.25 .642 .116 .660 8.25 .168 .716 .660 12.6 .271 .751 8.30 3.00 .421 4.09 .909 .375 3.00 .660 11.8 .168 .690 .330 93.4 .168 .669 .784 3.11 .703 1.14 5.67 .550 .414 24.2 .314 1.12 A. Boat Wks. Beach B. Harbor C. Del Monte Beach D. Del Monte Apts. E. U.S.N.P.G.S. F. Sewage Pipe G. Holiday Inn Zn Pb Cd 31.4 8.21 25.1 3.12 8.17 9.21 9.92 6.12 3.06 .492 4.83 .407 24.2 1.08 Figure 1. Collection sites. MENEII TII T Eil ——L . L L Holiday Inn HE L iin +———— Sewage Pipe. NIUSNPGS DelMonte Apts. Monten + —+ De AonteBeach, Bay t 1— in l Harbe . m 9 m — BoatiWks. Beach JMS ti ++— ut IH — EI. 4 iita titrititritit LIIIII LItN I 4 e 'igure 2. Trace metal levels in the sand from which the polychaetes were taken. *.. 16 N Cu 40 30 30 20 20 10 104 H HD — E G ABO C D A B 2006 Fe Mn 1800 40 1600 1400 10 0 H QL 100 ABCDELELG . Zn + Co 1 40 40 Amounts 3O Undetectable 20 20 10 10 + ++ n L BCDEE D Ai B A Boat Wks. Beach 40 B.Harbor 30 C. Del Monte Beach: 20 D.Del Monte Apts. 10 EHUSNPGS E Sewage Pipe A 3C D G. Holiday inn Sand lMetalllevelsin RRN.MitiL LLEig.2 Figure 3. Highest level of trace metals found in each species. 300 CU 3 40 6G 30 40 10 30 L 20 it A 10 MN Iiti LL L 20 A ) C 10 Ire 000 LEEE t BCDE S Z 300 E 0c 200 A L EE 100 C D A 40 60 umaamn H 3 540 2 O 30 L 20 ABC 10 CEuzonus dillonensis D.Euzonus mucronata ABCDE ravisia gigas ENephtys A. Nerinides acuta californiensis¬ B.Cinnifonmialspinabrangha HEigl3 Ni Figures 4-10. Trace metal levels in each species and the sand in which they were found according to geographical location. 20 O RC Nicke poo TE E Holiday Inn O t OE EE SewagePipe O LUSNPGSY Del Monte Apts 17 — — S DelMonte Beachl E Harbon U V If i Boat WksBeachln Mummn HMS m A.NERINIDES ACUTA B.CIRRIFORMIA SPIRABRANCHATIE C.EUZONUS DILLONENSS D.EUZONUS MUCRONATA: E.-TRAVISIA GICAS NEPHTYS CALIFORNIENSIS Mtitit Eig. 4 IIILIL 21 M — OA OO Copper Q O —I olidayinn 1 Sewage Pipey USNPGS Del Monte Apts t Del MonteBeach — — + 0 1 1 I O ++ O 1 it + CL Harbor — M — U BoatWks Beadh IMS ALINERINTDES ACUTA B.CIRRIFORMIA SPIRABRANCHA tt C. EUZONUS DILLONENSIS tiisiii HD-TEUZONUS MUCRONATA TF-TTRAVIS A GICAS ... E NEPHTYS CALIFORNIENSIS 4 Eig. 5 C 22 PPM C Manganese o 8 Se Holidaylnn O ... LILI — — Sewage Rpey USNPGS Del Monte Apts. i De Monte Beach: J — VO Harbon siti Boat Wks.Beach HMS L —A.—NERINIDES ACUTA¬ HH B. C IRRIFORMIA SP IRABRANCHA EUZONUS DILLONENSIS EUZONUS MUCRONATA HENITRAVISIA GIGAS +NEPHTYS CALTFORNTEN: m IAILLLIL LILLLMEig. 6 M — OO ron O T Holiday EILI SewagePipen USNPGSS Del Monte Aptsk TE Del MontelBeachs I - UE Habbon Boat Wks.Beach 5 HMS i MI 22 OO O — O L 9 O fi — — — ... D O — fi t — — NERINIDES ACUTA CIRRIFORMIA SPIRABRANCHA EUZONUS DILLONENSIS: EUZONUS MUCRONATA E. TRAVISIAGIGAS CALIFORNIENSIS NEPHTYS Eig.17 L Zinc Holidayllnn Sewage Piper USNPGS Del Monte Apts Del Monte Beach Harbot 1 Boat WksBeach t IMS + 7 ........ + ii n O m + A. NERINIDES ACUTA B.CIRRIFORMIA SPIRABRANCHA C-EUZONUS DILLONENSIS D.EUZONUS MUCRONATA EITRAVISIA GIGAS LF. NEPHTYS CAL FORNIENSIS Eig. 8 Q N 24 O O 85 PM Lead oo S Holiday Inn —0 1 + Sewage Pipe H — — USNPGS " +:: — Del MonteApts T — Del Monte Beach E JC Harbon ++ H 1 BoatWks. Beach D IN S ciast ronig e aaee . A-NERINIDES ACUTA CLEUZONUS DILLONENS IS EUZONUS MUCRONATA + TRAVISIA GIGAS NEPHTYS CALIFORNTENSS + i Eig. O ... Cadmium Holiday Inn Sewage Pipe USNPGS Del Monte Aptsk Del Monte Beach: Harbor Boat WksiBeach MS . PM o O I7 C O D 28 O +++1 Z Z V U + ++ 11 in at —1— A. NERINIDES ACUTA B.CIRRIFCRMIA SPIRABRANCHA C.EUZONUS DILLONENSIS D. EUZONUS MUCRONATA TRAVISIA GIGAS CALIIFORNIENSIS F. NEPHTYS Eig.10 26 L