Acknowledgements I would like to thank the entire staff of Hopkins Marine Station for their valuable assistance and encouragement, especially Dr. Malvern Gilmartin, Dr. John Martin, Mr. Robin Burnett, and the crew of the R/V Proteus. This work was made possible by grant GY 8950 of the Undergraduate Research Program of the National Science Foundation. C O O Abstract Plankton samples from the surface and from 10 meters depth were collected in Monterey Bay and analyzed for Ag, Zn, Cu, Cr, Mn, Cd, and Pb. The results showed a trend toward higher concentration at the air-sea interface. Possible mechanisms of concentration processes are discussed, with some consideration given to the ecological implications. 3 Introduction Because of the unique properties of the air-sea interface, it is surprising to find a paucity of information concerning the biology of this zone. It has been stated that chemical reactions in the ocean are "largely determined by phenomena which occur at interfaces" (Horne, 1969), and that almost all physical, chemical, and biological processes in the ocean depend on the ocean having a free boundary with the air (Wellander, 1961). Study of the air-sea interface would probably provide a good deal of information concerning the movement of heavy metals in the ocean. There is now good evidence that a significant portion of lead now entering the ocean does so via the atmos- phere; automobile exhaust has been found to contribute greatly to atmospheric lead pollution (Patterson, 1965, 1970; State of Cal. Air Resources Board, 1967; Mauchline and Templeton, 1964; Tatsumoto and Patterson, 1963). The interface is in constant exposure to such atmospheric fallout. Moreover, the physical and biological properties of the surface are such that materials are either concentrated or dispersed there. Materials in the surface layer are either heavier or lighter than sea water (Wilson, 1959); materials that are lighter float to the surface while heavier particles sink to deeper levels. Organisms facilitate both of these processes. They carry materials to the surface during their diurnal and seasonal migrations. 1 (Kuenzler, 1969 a,b). Also, small bubbles such as those produced by plants during conditions of high photosynthesis, and more importantly those produced by wave action, carry organic matter to the surface through a process of adsorption onto the surface of these bubbles. This process results in the formation of surface organic slicks(Riley, 1963; Riley, et.al, 1964; Siegel and Burke, 1965; Wilson, 1959; Blanchard, 1964; and Baylor and Sutcliffe, 1963). Materials move down- ward, again via plankton migrations, and also by elimination of waste products and detrital matter (Fowler and Small, 1967; Kuenzler, 1969 a,b; Martin, 1970; and Osterberg, et.al., 1963). It is likely that organisms living at or migrating to the surface of the ocean are important in both the concentration of heavy metals at the surface and transport to the ocean floor as a phase of the biogeochemical cycling of the various elements. Also, because heavy metalss are generally surface¬ active materials, and the surface of the ocean is a natural area where these types of materials accumulate (Blanchard, 1964; Baylor and Sutcliffe, 1963; Riley, 1963; Riley, et. al., 1964; Siegel and Burke, 1965; and Wilson, 1959), it is reasonable to suspect that heavy metals may be concen- trated at the surface of the ocean, relative to amounts found in the underlying water. The purpose of this study is to compare the concentrations of Pb, Mn, Ag, Zn, Cd, Cr, and Cu in organisms collected at the surface of Monterey Bay as compared to organisms collected at a depth of about 10 meters. Several different kinds of organisms were sampled and analyzed (calanoid copepods, the euphausid Euphausia pacifica, the ctenaphore Pleurobrachia bachii, and the siphonophore Velella lata); surface samples were designated as neuston, and subsurface samples labelled plankton. half hour taker W between 7:00 A.M. and 10.00 A.M. Boat spead was recorded and used in the calculatiens of relative biomass density While en board, the fresh samples were taken from the net and plaeed in aeid-eleaned (4-6 M Hno,) glaseware, cevered with aluminum foil, and stored en ke. In the lab, major groups of organisma vere separate This usually consisted of separating calaneid eoperod from P. bachij and medusae (especially Oetatina sp.) This was sasily done by passing the sample through a coarse (0.5 gm. mesh) net. The remaining sample was streined through a ring of phytoplankten netting, retai. the copeped segment of the sample. Excess water was remove nd wat weights recorded. All samples were driel at for at lesat 24 hours. Materials and Methods Basic means of collection were a neuston sled towed at the surface (David, 1965) anddsmall zooplankton net simultaneously towed about 10 meters below the surface. The two nets had openings of similar are (0.0h6 m for the sled, 0.061 m for the plankton net) and similar mesh (220.5 microns for the sled, 291 microns for net). The sled was either towed off to the side of the boat by means of a boom, or was trailed far enough aft so that the wake had little effect. Tows lasted from one-half hour to one hour. All inshore tows were taken between 7:00 A.M. and 10:00 A.M. Boat speed was recorded and used in the calculations of relative biomass density. While on board, the fresh samples were taken from the net and placed in acid-cleaned (1-6 N HNO,) glassware, covered with aluminum foil, and stored on ice. In the lab, major groups of organisms were separated. This usually consisted of separating calanoid copepods from P. bachii and medusae (especially Octatina sp.). This was easily done by passing the sample through a coarse (0.5 cm. mesh) net. The remaining sample was strained through a ring of phytoplankton netting, retaining the copepod segment of the sample. Excess water was removed and wet weights recorded. All samples were dried at 80°0 for at least 21 hours. Aliquots of the dried samples (less than l gm dry weight) were placed in acid-cleaned beakers and dissolved in concentrated HNO, (70-90%). Organic matter was destroyed using a modification of a method described by Middleton and Stuckey (in Christian and Feldman, 1970). The modified method consisted essentially of heating the sample with concentrated HNO,, evaporating the solution to dryness, redissolving the ash in acid, then adding a few ml of HoO, (30%) to oxidize any remaining organic material. A blank was included for each set of samples as a check for reagent contamination. After the sample was totally mineralized, the remaining ash was dissolved in concentrated HNO, and diluted to a standard volume with distilled H.0. The samples were then analyzed for Ag, Zn, Cr, Cu, Mn, Cd, and Pb on an atomic absorption spectrophotometer (model 303, Perkin-Elmer). 3/ damelty vis re Results Results from the analyses of three inshore and two offshore tows are presented in Table I and Figure I. The Vellela lata sample examined was taken from samples collected about 100 miles off the Oregon coast during a cruise of the R/V proteus in October, 1970. Highlights of the results follow. 1. The calanoid copepod samples collected at the surface had higher concentrations of Ag, Zn, Pb (p =.20; see Table II), and Cr (p = .10) than samples collected at 10 meters depth, while Cd concentration was higher in the plankton (p -.10). 2. Offshore plankton copepods had over twice the Pb concen- tration (10.37 £ 1.61 ppm vs. 1.28 - 1.12 ppm; see Table I) as that found in the inshore plankton copepod samples (p =.01). 3. Euphausia pacifica samples at the surface and at 10 m. showed no significant difference (all p values greater than 20) in heavy metal concentration except for Pb, for which the plankton samples had about twice the neuston Pb concen- tration (5.39 I .56 vs. 2.15 -.32, p -.05). 1. There was no significant difference measured for any heavy metal concentration between neuston and plankton samples of Pleurobrachia bachii (all p values greater than.30). 5. As the relative density of copepods in the surface zone decreased in a series of runs, a corresponding increase in C plankton density was recorded (see Figure II). 6. There is an inverse correspondence between Od concen- tration and relative biomass density in plankton copepods. and a positive correlation between density and Cd concen- tration in neuston copepods. Also, there are inverse correlations for Pb and Zn vs. density in neuston copepods. Discussion Results of this study do not show with certainty a process of heavy metal concentration at the air-sea interface. The probable reason for this is that the organisms sampled were members of a vertically migrating population rather than members of distinct neuston and plankton communities. Despite this, however, there was a trend toward higher concentrations in the surface copepod samples over those samples collected at 10 m. This strongly suggests that some heavy metals are concentrated at the interface. It is interesting that this trend is in the opposite direction to that found by Martin (1970), who reported higher heavy metal concentrations in copepod sampled at depths greater than 100 m. than in samples from surface waters. This apparent disparity may be related to the depth of sampling. The purpose of this study was to was to analyze movements of heavy metals at the air-sea interface and immediate underlying waters, while Martin was looking at larger scale movements. His "surface water" sampling zone included both zone sampled in this study. Despite this, there is fairly good agreement between values reported here and those given by Martin (Table Il1 Additional evidence for the phenomena of surface concen¬ tration comes from the results of Baxter (1971) on these same copepod samples for analysis of DDT. He noted that the surface samples had a greater concentration of DDT than did the plankton samples. Also, Marion (1971) reported that O 320 the DDT concentrations in Hermissinda crassicornis collected from floats were 2.3 times higher than those collected pilings, 0.5 to 7 m. below the surface. Offshore planktonic copepods had significantly more Pb than the inshore planktonic copepods, for reasons as yet unknown. An anomalous set of data resulted from analysis of the Euphausia pacifica samples collected during tow 15. For six of the metals analyzed there was no significant differ- ence between the surface samples and those collected at 10 m. Moreover, Baxter (1971) found that the two samples had the same DDT concentrations. This homogeneity for E. pacifica might be expected because this organism is a strong vertical migrator, and may move from a depth of several hundred meters up to the surface. However, the 10 m. samples contained about twice the Pb concentration of the surface samples. Physiological differences between the populations are not considered likely because they would tend to produce more than a difference in the concentrati on of just one metal. It is quite possible that, because only two samples were analyzed, and that the concentrations of Pb measured were at the limit of sensitivity for measurement by atomic absorp- tion, the statistical test for this particular case was not valid. The neuston and plankton samples of Pleurobrachia bachii showed no significant differences with respect to heavy 32 metal concentration. This is not surprising. P. bachii is capable of strong and rapid vertical migrations; they will move down when surface conditions are rough, and also when light intensity is high (Hyman, 1940). Also, because their surface area to volume ratio is minimal, P. bachii does not concentrate high levels of heavy metals by surface adsorption. Therefore, if differences between surface and subsurface samples do exist, they are more difficult to observe. It is reasonable to expect an inverse relationship between plankton vs. neuston density for the copepod samples because copepods exhibit vertical migrations. The results presented in Figure II indicate that both the neuston and plankton samples were at least partly composed of a vertically migrating population. But it is likely that some organisms spend most of their time at the surface, while other organisms seldomly leave the subsurface zone, because copepods were always found in both zones during simultaneous tows at the surface and 10 m. Copepods collected at the surface when relative density was low are probably those organisms having a long residence time on the surface. Similarly, samples collected at 10 m. during low density conditions contained those copepods that rarely migrate to the surface. The existence of either of these populations is masked when density is high. This is the type of subdivision one might expect within 322 a large population; while the majority of organisms will show an average amount of migration, some individuals will be skewed on either the high or low side of the mean distance and frequency of migration. There is evidence for some of the differences occurring between individuals in a copepod population. It has been noted that female copepods come to the surface to lay eggs, and also that females migrate more regularly than males (Marshall and Orr, 1955). Bainbridge (1952, in Marshall and Orr) observed that some copepods may inhabit only the top 30 cm. of water, while others seldoml migrate that close to the surface. It is reasonable to suspect that some copepods spend much of their time at the surface, while others more exclusively inhabit the subsurface zone. If there is a difference in metal concentration between the surface and subsurface, those organisms that spend the greatest amounts of time at each zone should exhibit those differences most clearly. There is good evidence that this is true. Relative density is inversely related to time; a low density value gives some measure of the length of time certain organisms spent in one zone. Figure IIIdemonstrates some relationships between metal concentration and residence time, as measured by relative density. From these relationships one can gain some knowledge as to the depths at which various metals are concentrated. 322 The relationships for Cd concentration vs. density Fiqve lLb) are especially interesting. In the plankton samples, concentration increases as density decreases; that is, concentration increases as residence time increases. The opposite is true for neuston copepods; concentration increases as density increases. This type of relationship might occur if Cd is in higher concentration in subsurface waters than at the surface. Organisms that remain at depth for a longer period of time would tend to concentrate the metal through adsorption and feeding. Also, as copepods that have spent some time in the subsurface waters migrate to the surface, they carry with them relatively high Od loads. So concentra- tion of Od in neuston increases while density also increases. It is interesting to note that Cd is the only metal tested that showed a higher concentration in the subsurface samples. Zn and Pb concentrations in neuston copepods show the same type of relationship demonstrated by Od in the plankton. There are no corresponding relationships in the subsurface samples for these metals, however. The probable reason for this lack of correspondence is that the downwardly migrating copepods diffuse throughout several meters of the subsurface waters so that there is no clear gradient. These results do support the hypothesis that at least Zn and Pb are concentrated at the air-sea interface. Conclusion The results presented here favor the hypothesis that certain heavy metals are concentrated at the air-sea interface relative to concentrations found in underlying waters. Problems encountered in sampling, analysis, and population structure of the plankton tended to mask these differences. The implications from this surface concentration effect may be significant. It is likely that organism feeding on this surface layer, such as larval fish and birds like skimmers and phalaropes, may be receiving a relatively high metal load in their food. Metals concentrated at the surface may feed directly into both aquatic and terrestrial food webs. Also, because of the chemical reactions that place at the surface, the concentration effect at the interface may play an important role in the biogeochemical cycling of these elements. References Cited Baxter, K.G., (1971), "Comparison of DDT residue values of planktonic organisms at the air:water interface and subsurface levels of Monterey Bay", (Unpublished MS on file at Hopkins Marine Station Library) Baylor, E.R. and W.H. Sutcliffe, Jr., (1963), "Dissolved organic matter in sea water as a source of particulate food", Limnol. Oceanog. 8: 369-371. Blanchard, D.C., (1964), "Sea-to-air transport of surface active material", Sci 116: 396-7. Christian, G.D. and F.J. Feldman, (1970), Atomic Absorption Spectroscopy. Wiley-Interscience, N.Y. Cox, L.J., (1971), "DDT residues in sea water and particulate mater in the California current system", Fish. Bull. (in press) David, P.M. (1965), "The neuston net; a device for sampling the surface fauna of the ocean", J. Mar. Biol. Assoc, U.K. 15: 313-320. Fowler, S.W. and L.F. Small (1967), "Moulting of Euphausia pacifica as a possible mechanism for vertical transport of Zn-65 in in the sea", Int. J. Oceanol. Limnol 1: 237-215. Horne, R.A. (1969), Marine Chemistry, wiley-Interscience. N.Y. 568p. Hyman, L. (1940), The Invertebrates, V.1. McGraw-Hill, N.Y. 726p. Kuenzler, E.J. (1969a), "Elimination of I, Co, Fe, and Zn by marine zooplankton", pp. 162-173 in D.J. Nelson and F.C. Evans, (ed.), Symposium on Radioecology. photocopy 324 reproduced by Atomic Energy Commission. 774p. (1969b), "Elimination and transport of Co by marine zooplankton," pp. 483-492. ibid. Marion, R. (1971), "Assimilation and transfer of DDT residues in a hydroid-nudibranch-tetribranch food chain) (Unpublished MS on file at Hopkins Marine Station Library). Marshall, S.M. and A.P. Orr, (1955), The Biology of a Copepod, "Calanus finmarchicus". Oliver and Boyd, Edinburgh. 188p. Martin, J.H., (1970), "The possible transport of trace metals via moulted copepod exoskeletons", Limnol Oceanog. 15: 756-761. Mauchline, J. and W.L. Templeton, (1964), "Artificial and natural radioisotopes in the marine environment", Oceanog. and Mar, Bio. A. Rev. 2: 229. Osterberg, C.L., A.G. Carey, Jr., and H. Curl, (1963), "Acceler- ation of sinking rates of radionuclides in the ocean", Nature 200: 1276-77. Patterson, C.C., (1970), "Lead", in D. Long, ed., Impingement of Man on the Ocean. preprint. —-------------, (1965), "Contaminated and natural lead environments of man", Arch. Env. Health 11: 311-360. Riley, G.A., (1963), "Organic aggregates in sea water and the dynamics of their formation and utilization", Limnol. Oceanog. 8: 369-381 Riley, G.A., P.G. Wangersky, D.V. Hemert, (1964), "Organic aggregates in tropical and subtropical surface waters of the North Atlantic Ocean", Limnol. Oceanog. 9: 546-50. Siegel, A., and B. Burke, (1965), "Sorption studies of cations on 'bubble-produced organic aggregates' in sea water". Deep-Sea Res. 12: 789-796. State of California, Air Resources Board, (March, 1967), Lead in the Environment and its Effect on Humans. 81 p. Tatsumoto, M., and C.C. Patterson, (1963), "The concentration of common lead in sea water", pp. 71-89 in J. Geiss and E.D. Goldberg, ed., Earth Science and Meteoritics. North Wellander, P., (1961), "Coupling between sea and air", pp. 4O1- 110 in M. Sears, ed., Oceanography, Horn-Shafer, Baltimore. 654 p. Wilson, A.T., (1959), "Surface of the ocean as a source of air¬ borne nitrogenous materials and other plant nutrients", Nature 184: 99-100. C Table 1-- Mean values obtained for Ag, Zn, Cu, Cr, Mn, Cd. and Pb concentration in samples collected during five tows in Monterey Bay. The standard error was computed as follows; SE - N-1 32 8 S 10 14 - ++ 40 44 4. 1+ .9. .O . — .O O O 4 14R + — + + 4 —. N- N- o 50 o + + + — . ORE • O V 8 +0 + + + +6 40 1+0 10 a N o O OO + — 4. ++ +N + +0 . .0 O CE + + +: + N +N — — — .J . SO OO Do O + + 1 + + + * 3 O 2 0 o . 6P 86 6 0 85 28 OS g o 0 12 3 R 1+4 + 1+ 0 4N +. • .O .0 5 O 400 4 +N + 140 +N e 8 + E L N 8 + +N + + + . O 8 * O • O O NO 55 O ++ R ++ +N + 40 +0 + 40 4 9e +. 0 +e in O O O OO +O +0 ++ +0 +140 40 +0 H — — - NO . O • O OE 0 + 140 40 + +0 1 40 + ON 5 —0 A • O O O 0 4 40 + 4 4 +N +0 +0 — i OO OO o 5 5 00 320 N 00 + + 4 ON 28 +O + N ES 40 + +. O. P 10 + + . O O +0 45 —0 —0 ++0 5 —0 • + N + o . O 0 O Table #2 - The probability that the two populations compared are from the same general population, as computed by the "t" test, ie. t =X: - 22 65 11 n-1 33 O Tabl Populations compared Nueston copepods vs. Plankton copepods Nueston E. pacifica Plankton E. pacifica Nueston P. bachii vs. Plankton P. bachii Plankton inshore opepods vs. Plankton offshore opepods 33, Cr Ag Zn Cu Mn Cd Pb .20 .20 .10 .60 .40 .10 .20 .80 .70 .30 .80 .20 .80 .05 .80 .90 .60 .80 .50 .50 .30 .10 .O1 .30 .80 .70 .30 90 O e 0 0 Table III-- Comparison of concentrations of Cu, Mn, Zn. and Pb between results of this study and those reported by Martin (1970). 33. O Table III this study a) all copepods n = 22 b) neuston copepods n = 11 c) plankton copepods n = 11 Martin (1970) a) surface copepods n = 10 b) all copepods n = 22 Szabo (1968) copepods Cu 19.71 t5.26 23.29 + 10.30 16.12 + 3.67 38 11 30 Mn 7.91 + 1.21 6.64 +1.63 9.19 + 1.77 23 29 11 Zn + 11 315 + 74 191 + 16 220 130 266 Pb 18.18 + 8.96 29.96 -17.51 6.19 + 1.39 39 19 —— 35 0 C Figure Fl - Species composition and towing conditions for the 11 runs in Monterey Bay. Abundance scale is: +++ most abundant; ++ frequently observed; + seldom observed. Circles indicate sewer effluents; corresponding numbers show treatment (ie. primary, secondary, etc.). 33 33 San Lorenzo FIGURE 1 River SANTA CRUZ Q Tow l-3 runs /7 1./29 15-0915 Calm-overcast alnoid oeos ++ p. bachii Tow 2-1 run ocoo-C830 5/6 Overcast,cold,wind Plankton: (10-15 M) euston: Calanoid opepods + Calnid oepod ++ achi++ surface) +++ bachi val shrimp ++ a rval shrimp ++ Ta: larval crab (zoa) + arval rab Elkhorn Leptomedusae + Gammarid amphipod + V Beroe forskali Slough larval fish+ phausid+ 3-3 runs To 5/1 30-1030 alm,overast Plankton: Galanoid copero Clnd oepods + Neus " +++ p. bachi + +++ P. bac rval crab (megalops) amphapods + ow -3 runs Salinas River 7 5/11 o900-1300 atly loudy,choppy Plankton:Calanoid copdpods + ton:alanoid opepods ++ Hyperid amphapods + Siphonophores + ral fish yperid amphapods + Euphausid P. bac! i + Tomopterus runs 57 5/21/7 TOW2 o030-02 101/3 ny,choppy,cold rons ia +++ Meuston: C. P rval crab (megalops sh (small) MONTERE Siphonophores + 6 Medusae + Tow 15 (con);0600-0700 Calanid opepo ndy,choppy,cold ifia + plankton: Neuston: Calanoid copepods p. bachii (10 M) crab larve (megalops) Calanoid copopods ica oa squid (juvenile) hapods + D7 lankton-10 M: Carmel River Calanoid copepods J iphonophores + Medusae . bachi T0W 5 larval fish+ O Figure +2 - Neuston copepod relative density vs. planktoi copepod relative density for tows 9.O -10.0 -11.0 O.O FIGURE II 0 —TOW -TOW 2 X -TOW 4 8.O -7.0 log plankton density (g/m- 33 O Figure III a-- Graph of log relative density vs. log concentration of Pb and Zn in calanoid copepod samples collected at the surface. Figure III b-- Graph of log relative density vs. log concentration of Od in plankton copepod samples and in neuston copepod samples. C 6.0 5.O 4.0 3.0 2.0 -11.0 FIGURE IIa Zn— Neuston ° Pb— Neuston —10.0 —9.0 log density (g/m3) — 2.0 1.5 1.0 11.0 10.0 7.0 9.O 8.0 log density (g/m3) FIGURE Tb Cd o- Plankton •-Neuston 00 0 340