Biochemical correlates of dissolved mercury uptake by the oyster Ostrea edulis

From previous work, the equilibrium concentration factor for dissolved mercury in the digestive gland of Ostrea edulis Linnaeus was found to be three to four times higher than that in the gills. In the present study, an analysis of soluble protein revealed values of 49.3±14.2 mg g wet tissue^-1 for the digestive gland and 0.7 ±0.1 mg g wet tissue^-1 for the gills. Starvation significantly reduces the soluble protein level of the digestive gland to 31.1±6.4 mg g^-1 and that of the gills to below the limit of detection. These results suggest that the difference in concentration factors between the gills and digestive gland may be based on a quantitative difference in macromolecular binding sites. However, the uptake of dissolved mercury over a period of 48 h was considerably greater in the gills, so that although the soluble protein content of the tissue may influence the final concentration factor, it does not appear to affect the rate at which this equilibrium is achieved. A more detailed investigation of the mechanism of dissolved mercury uptake by oyster gills has been carried out using isolated tissues. The process is inhibited by 5mM 2–4 dinitrophenol, by the absence of a readily metabolizable substrate (dextrose) in the uptake medium, and by 30mM K⁺. The effect of K⁺ necessitated further investigation with a specific inhibitor of K⁺ transport. Strophanthin G (ouabain), at a concentration of 0.01 mM, caused a significant increase in mercury uptake.     

Influence of temperature and salinity on the uptake,distribution and depuration of mercury,cadmium and lead by the black-lip oyster Saccostrea echinata

Saccostrea echinata (Quoy and Gaimard) were exposed to 10 g 1^-1 of either mercury, cadmium or lead at 30 °C, 36S; 30 °C, 20S; 20°C, 36S and 20°C, 20S for 30 d and were then transferred to clean seawater for a further 30 d to depurate. Specimens were removed at regular intervals during the exposure and depuration periods, dissected into gills, mantle, visceral mass and adductor, and analysed for the appropriate metal by atomic absorption spectroscopy. Mercury was concentrated more than the other metals in all tissues under all conditions. Cadmium uptake was greater than lead in all tissue in the high-temperature experiments, whereas both metals were concentrated to similar extents at low temperature. The gill tissue generally accumulated the greatest amount of all 3 metals, whilst the adductor concentrated the least amount. At both salinities, mercury and cadmium accumulation by all tissues was significantly greater at the higher temperature whereas lead uptake was only marginally increased. The accumulation rates of mercury at high temperature were significantly greater in all tissues at low compared with high salinity, whereas at low temperature, differences were not significant. Accumulation rates of cadmium and lead in the majority of tissues examined were significantly greater in lowsalinity water at both temperatures. In general, lead was lost the most rapidly from oyster tissues, followed by mercury and then cadmium. The residence times for mercury and cadmium differed significantly between tissues, with the gills showing the highest turnover rate. In contrast, residence times for lead were similar between tissues. Losses of all 3 metals from oyster tissues were not obviously influenced by temperature and only mercury losses differed significantly between salinities.     

Microprofiles of oxygen in epiphyte communities on submerged macrophytes

Mussels (Mytilus edulis) transferred in net bags from clean to chronically mercury polluted water readily accumulated mercury during an exposure period of three months. Growth of the transplanted mussels had a diluting effect on the mercury concentration, but the absolute weight of mercury uptake increased throughout the entire period, though there was a tendency for decreased efficiency of the removal of mercury per liter of water filtered by the mussels. Mussels were also translocated from polluted to clean (laboratory) water to depurate mercury. The biological half-lives of mercury was 293 d for M. edulis from the chronically polluted area in contrast to only 53 d for mussels from a temporary massive mercury polluted area near a chemical deposit. In both cases about 75% of the total mercury in the mussels was inorganic, and it is suggested that both inorganic and organic mercury species were immobilized in mussels from the long-term mercury polluted area, whereas the immobilization capacity was exceeded in the short-term mercury exposed mussels near the chemical deposit. Very slow elimination of mercury was observed in the deposit-feeding bivalve Macoma balthica from the chronically polluted area, and about 6% of the total mercury was methyl-+phenyl-mercury. This is more than three times lower than found in M. edulis from the same collecting site. A pronounced difference in the mercury speciation (i.e., total mercury, total organic mercury, methyl-mercury and phenyl-mercury) in M. edulis from the two mercury polluted areas is thought to reflect the different character of the mercury pollution in the two areas.     

Allometry of gill weights,gill surface areas,and foot biomass δ<Superscript>13</Superscript>C values of the chemoautotroph–bivalve symbiosis<Emphasis Type="Italic"> Solemya velum</Emphasis>

The protobranch bivalve Solemya velum Say, 1822 has large gills, which harbor chemolithoautotrophic bacteria that supply the majority of the clams organic carbon. A substantial portion of the CO₂, O₂, H₂S, and other nutrients necessary for symbiont autotrophy and host heterotrophy are acquired from the environment through the gills, whose large size may be necessary to facilitate the acquisition of sufficient O₂ from S. velums habitat to meet the combined demands of the host and symbionts. Large gills may also result in an oversupply of CO₂, which may in turn be responsible for the isotopically depleted ¹³C values observed in S. velum biomass (–31 to –34). Alternatively, gill hypertrophy may simply be an adaptation to house a large population of symbionts adjacent to their environmental source of dissolved gases and other nutrients. To better understand gill function in this symbiosis, gill weights, gill surface areas, and foot ¹³C values were measured as a function of total weights. S. velum gill weights were found to be a substantial portion of total clam weight, averaging 38% of wet weight, compared to nonsymbiotic protobranch bivalves Yoldia limatula Say, 1831 (5%) and Nucula proxima Say, 1822 (11%). Gill weights are a smaller percentage of total weight in larger individuals; the allometric equation for gill weight (G) as a function of total weight (M) is G=0.26M^0.85. Dry weights scale similarly. Gill surface areas are immense; the average gill surface area measured was 107 cm² g^–1 total soft tissue wet weight, the highest value for any marine invertebrate. Gill surface area (SA) also scales with size (SA=69.8M^0.85). When gill surface areas were calculated with respect to gill wet weights, they did not scale with size. The ¹³C values do not scale with size either, consistent with high rates of CO₂ supply at all sizes. Extraordinarily high rates of CO₂ supply relative to demand are supported by a model for CO₂ delivery based on Ficks law and the allometric relationship between surface areas and total weight, consistent with a role for large gill surface areas in the generation of isotopically depleted tissue ¹³C values.Communicated by J. P. Grassle, New Brunswick     

Transbranchial potentials and ion fluxes across isolated,perfused gills of Uca rapax

Transbranchial potentials (TP) and sodium or chloride fluxes were measured in an apparatus designed for the simultaneous perfusion of eight isolated gills of Uca rapax. In anterior gills perfused with U. rapax–saline (US) the TP varied almost linearly from-7.5 to +10 mV inside, and in posterior gills from +2 to-8.5 mV (inside), on exposure to salinities ranging from 8.7 through 52, i.e. 25 to 150% seawater (100%=34.6 S). Sodium influx and efflux in anterior gills exposed to US, 8.7 or 43.3 S (0.7 to 4.0 mmol h^–1 g^–1 dry wt) were always greater than in posterior gills (0.5 mmol h^–1). The chloride fluxes were slightly smaller than sodium fluxes in anterior gills, while in the posterior gills the chloride influx (2.8 to 4.6 mmol h^–1) was always larger than chloride efflux (0.6 to 1.1 mmol h^–1) or the sodium fluxes. At least three ion-transport mechanisms may be present in these gills: (1) an internal ( = basolateral), ouabain-sensitive Na⁺, K⁺ pump, restricted to anterior gills; (2) a furosemide-sensitive Na⁺, K⁺, 2Cl^– (plus water) transporter, apparently restricted to posterior gills, and (3) a Na⁺ exchanger (and possibly other as yet unidentified ion transporters, as suggested by large increases of the chloride influxes caused by amiloride), probably located on the apical membranes of the epithelial cells of both gill types. The differential selectivity of the gills of U. rapax for sodium or chloride may limit the transbranchial movements of either ion, without a reduction of the overall permeability of these crabs.Communicated by N.H. Marcus, Tallahassee     

Scanning electron microscopy study of the gills,scales and erythrocytes of Anabas testudineus upon exposure to chlorpyrifos

The present study was undertaken to assess the toxicity of sublethal concentrations (125, 250, and 375 µg L^?1) of chlorpyrifos on Anabas testudineus for 21 days. The morphological changes on the gills, scales, and erythrocytes of A. testudineus were observed using scanning electron microscopy. Gill alterations included highly active mucous cells, epithelial hyperplasia, and fusion of secondary lamellae. The scales showed damaged lepidonts. Oozed out cytoplasmic content and lobopodial projections were observed in the erythrocytes after exposure to chlorpyrifos.     

Bioenergetics and survival of the marine snail Thais lima during long-term oil exposure

The carnivorous snail Thais lima was fed Mytilus edulis during a 28-d exposure to the water soluble fraction (WSF) of Cook Inlet crude oil. The LC-50 of T. lima declined from >3000 ppb aromatic hydrocarbons on Day 7 to 818±118 ppb on Day 28. The LC-50 of M. edulis declined from >3 000 ppb aromatic hydrocarbons on Day 7 to 1 686±42 ppb on Day 28. Predation rate declined linearly with increasing aromatic hydrocarbon concentration up to 302 ppb; little predation occurred at 538 ppb and none at 1 160 or 1 761 ppb. Snail absorption efficiency averaged 93.5% and did not vary as a function of WSF dose. Total energy expenditure (R+U) increased at 44 ppb aromatics and declined at lethal WSF exposures. At sublethal WSF exposures, percentages of total energy expenditure were: respiration (87%), ammonia excretion (9%) and primary amine loss (4%). These percentages did not vary as a function of WSF dose or time. Oxygen:nitrogen ratios were not affected by WSF concentration or time and indicated that T. lima derived most of its energy from protein catabolism. The uptake of aromatic hydrocarbons into the soft tissues of snails and mussels was directly related to the WSF concentration. Naphthalenes accounted for 67 to 78% of the aromatic hydrocarbons in T. lima and 56 to 71% in M. edulis. The scope for growth was negative above 150 ppb WSF aromatic hydrocarbons and above 1 204 ppb soft-body aromatic hydrocarbons. These snails were physiologically stressed at an aromatic hydrocarbon concentration which was 19% of the 28-d WSF LC-50 (818±118 ppb) and/or 48% of the 28-d LC-50 of soft tissue aromatics (2 502 ppb).     

Effects of sublethal concentrations of mercuric chloride on ammonium-limitedSkeletonema costatum

The effect of sublethal additions of mercuric chloride on the marine diatomSkeletonema costatum (Grev.) Cleve grown in NH₄-limited chemostats and batch cultures was assessed. In short-term Hg exposure experiments (up to 5 h), the effect of Hg on ammonium uptake rates was studied by simulatneously perturbing the culture with 5 M NH₄ Cl and Hg concentrations ranging from 0.04 to 5.52 nM HgCl₂. The threshold of Hg toxicity occurred between 1.8 and 3.7 nM, based on a decrease in ammonium uptake rates. When the NH₄-limited culture was starved of ammonium for 30 h, the threshold of Hg toxicity decreased about an order of magnitude to 0.2 nM. In long-term Hg exposure experiments (679.5 h), NH₄-limited continuous cultures were semi-continuously exposed to 0.37 and 3.68 nM HgCl₂. After 4 days, the cell density in the Hg-treated chemostats began to drastically decline. After about 16 days these populations recovered, even though Hg additions continued. At the end of the experiment (26 days), cell densities had reached the levels observed at the beginning of the experiment. The reason for the recovery is unknown, but several possibilities are discussed. Ammonium uptake rates determined during the time-course of this long-term Hg exposure, indicated that these NH₄-limited cultures exhibited a significant loss in their ability to take up ammonium at low concentrations (e.g. 1 M). Thus, mercury pollution may seriously decrease the ability of a species to utilize the limiting nutrient during periods of seasonal nutrient limitation.     

Effects of temperature,nutritional factors and salinity on the uptake of L-methionine by the Sydney rock oyster Saccostrea commercialis

The effects of physiological and nutritional factors and of temperature on the uptake of L-methionine by Saccostrea commercialis were investigated on cultured rock oysters from Port Stephens, New South Wales, Australia, in 1983. Optimum conditions for L-methionine accumulation were 30°C, 18 h exposure and concentrations of other amino acids less than that of L-methionine. The uptake of L-glycine was inhibited by L-methionine in a reciprocal manner. There was no effect of salinity on the accumulation of L-methionine by acclimated oysters. During the latter investigations the range of osmoconformity was found to be 15 to 45 S. Oysters take 2 d to conform to new media osmolarities after salinity changes of 15. Amino acid supplements in oyster diets should be kept within the same order of magnitude to reduce inhibition of uptake.     

Effect of copper on ion- and osmoregulation in the shore crab Carcinus maenas

The uptake and effect of dissolved copper on regulation of hemolymph osmolality and Na⁺, K⁺, Cl^-, Ca⁺⁺, and Mg⁺⁺ concentrations in the shore crab Carcinus maenas (L.) were determined at 400 mOsm (=14 S) ambient salinity. One mg Cu l^-1 resulted in 50% mortality in 11 to 22 d; the highest sensitivity was observed around the moulting period. 0.25 and 0.5 mg Cu l^-1 were not lethal after a onemonth exposure. Ten, 1, and 0.5 mg Cu l^-1 altered regulation of osmolality, Na⁺, K⁺, and Cl^- concentrations, while 0.25 mg Cu l^-1 did not. Exposure to 1 mg Cu l^-1 reduced hemolymph osmolality and Na⁺, K⁺, and Cl^- concentrations to 80 to 90% of controls within 4 to 6 d and no further reduction was observed during a 21-d exposure. The effect of various copper concentrations on these four parameters were almost identical and the highest sensitivity was observed around the moulting period. Hemolymph calcium levels increased 20 to 80% in crabs exposed to 1 mg Cu l^-1, while they decreased 20 to 30% in crabs exposed to 0.5 mg Cu l^-1. Prolonged exposure to copper caused 20 to 70% reductions in hemolymph magnesium levels. Crabs exposed to 0.5 mg Cu l^-1 for 29 d accumulated copper in hepatopancreas, gills, carapace, heart, testes, and hypodermis, but not in muscles and hemolymph. Increased copper levels in crabs exposed to 0.25 mg Cu l^-1 were observed in hepatopancreas, gills, and carapace only. The present results suggest that effects of copper on ion regulatory processes in part explain why the toxicity of copper towards euryhaline invertebrates increases at low salinities.     

Arsenic in garden soils and vegetable crops in Cornwall,England: Implications for human health

Total concentrations of arsenic in surface (0–15cm) garden soils in the historical mining area of Hayle-Camborne-Godolphin, Cornwall, England are large and range widely (144–892 g/g). Amounts of water soluble and acid-fluoride extractable soil arsenic are significantly correlated with total content.Examination of 6 salad and vegetable crops grown in 32 gardens has shown arsenic concentrations in the edible tissues to be only slightly elevated. There were strong correlations between arsenic in beetroot, lettuce, onion and peas and soil arsenic (total, water soluble and acid extractable). Regression equations have been calculated. Ridge regression analysis applied to test the importance of other soil variables has shown both iron and phosphorus to influence the uptake of arsenic.Arsenic in all the vegetables sampled was below the statutory limit in the U.K. of 1 mg/kg fresh weight. Implications for health should be assessed in relation to other exposure routesvia water, air and directly ingested dust and soil.     

Accumulation and depuration of mercury in the American oyster Crassostrea virginica

To study the kinetics of mercury uptake in oysters, adult Crassostrea virginica (Gmelin) were held in seawater containing 10 g mercury/l (ppb) or 100 g mercury/l (ppb), added in the form of mercuric acetate, for 60 days. Mercury concentration in tissues was determined by analysis of individually homogenized oyster meats, using wet digestion and flameless absorption spectrophotometry. After 45 days, average mercury tissue concentration was 140,000 g mercury/kg tissue (ppb) and 28,000 g mercury/kg tissue (ppb) in the 100 ppb and 10 ppb experimental groups, respectively. After this time, concentrations dropped sharply, probably due to spawning. Clearance of mercury from tissue was studied by exposing treated adults to estuarine water (with no additions) for 30 days (100 ppb group) and 160 days (10 ppb group). Tissue concentrations in the 100 ppb mercury environment group declined from 115,000 to 65,000 ppb, and those of the 10 ppb group declined from 18,000 to 15,000 ppb, in 18 days; there-after, no further decline occurred in either group. Oysters accumulated mercury 1,400 times and 2,800 times above the environmental concentrations of 100 and 10 ppb mercury, respectively. Total self-purification was not achieved over a 6 month cleansing period.     

Identification of protein biomarkers of mercury toxicity in fish

Bioaccumulative metals such as mercury are found in increasing amounts in fish and their consumers. In the region of the Madeira River, in the Brazilian Amazon, mercury (Hg) is a predominant contaminant in the aquatic ecosystem. There is therefore a need to find specific biomarkers of mercury toxicity in fish to monitor contaminations. Here, mercury-bound proteins were identified in the liver tissues of fishes Mylossoma duriventre and Brachyplatystoma rousseauxii. Mercury was quantified in liver tissue, pellets and protein spots by graphite furnace atomic absorption. Proteins were fractionated by two-dimensional polyacrylamide gel electrophoresis and identified by mass spectrometry with electrospray ionization. We identified nine proteins linked to mercury and that presented biomarker characteristics of mercury. Among the proteins identified, isoforms of parvalbumin, ubiquitin-40S ribosomal protein S27a, brain-specific angiogenesis inhibitor 1-associated protein 2-like protein 2 and betaine–homocysteine S-methyltransferase 1 are notable for having the molecular function of binding to metallic ions.     

A kinetic and autoradiographic study of the direct assimilation of amino acids and glucose by organs of the musselMytilus edulis

The uptake of amino acids and glucose dissolved in sea water by different parts of common mussels (Mytilus edulis, L.) is studied from the first minutes up to 1 week. Autoradiography and donble countings on¹⁴C and²H labelled samples show that, in the first hour, the gills and mantle can concentrate several hundred times the dissolved nutritive molecules from very dilute solutions, whereas the digestive tract is not involved significantly during the first hours. An important -amylasic activity has been detected in the gills. The gill epithelium shows a strong positive reaction with mixtures used for the histochemical detection of chymotrypsin. This suggests that the digestion of small particles as well as the absorption of dissolved food might be initiated on the surface of the palleal-gill areas, and completed later in the hepatic caeca.     

Subcellular distribution of zinc and cadmium in the hepatopancreas and gills of the decapod crustacean Penaeus indicus

The decapod crustacean Penaeus indicus accumulated Cd and Zn in different subcellular compartments of hepatopancreas and gill cells. Most of the Cd and part of the Zn accumulates within the soluble fraction of the cells, while the remainder of the Zn is found in insoluble inclusions, associated with P, Ca, Mg and Si in B-, F- and R-cells in the hepatopancreas, and haemocytes, nephrocytes and epithelial cells in the gills. No presence of Cd was observed in metal-rich inclusions in any cell analysed. Metallothionein-like proteins (MTLP), analysed by differential pulse polarography, were present in the hepatopancreas (12–18 mg g⁻¹) and gills (7–8 mg g⁻¹) of metal-exposed prawns. Binding to MTLP is the detoxification mechanism for cadmium, while the detoxification of zinc involves both binding to MTLP and incorporation into insoluble metal-rich inclusions.     

Sodium regulation in the shore crab Carcinus maenas as related to ambient salinity

The activity of Na–K-ATPase was measured in crude homogenates prepared from various organs (leg muscle, pincer muscle, heart, testes, digestive gland, hypodermis, gills 1–9) of shore crabs, Carcinus maenas L., acclimated to salinities ranging between 10 and 50 S (in steps of 10 S). In all salinities tested, Na–K-ATPase activity was highest in posterior gills 7–9 (10–12 mol P_i mg protein^-1 h^-1), followed by anterior gills 1–6 (ca. 2.5 mol P_i mg protein^-1 h^-1) and the other organs (in most cases far below 2mol P_i mg protein^-1 h^-1). In gills only, Na–K-ATPase activity was salinity-dependent, with the highest values in the lowest salinities and vice versa. In gills 7–9, Na–K-ATPase activity was increased more than threefold following a reduction in salinity from 50 to 10 S. Na–K-ATPase activity, expressed as percentage of total ATPase activity, amounted to 60–80% in gills, about 60% in hypodermis and 20–40% in the other organs. Ouabain, a specific inhibitor of Na–K-ATPase activity, reduced serum osmolalities in crabs kept at 9–10 S only when injected into the hemolymph (1 and 5 · 10^-5 M), but had no effect when dissolved in ambient water (10^-4 M). The results obtained underline that crustacean gills are the main organs for ionic regulation, and confirm the hypothesis of the central role of the Na–K-ATPase in active Na uptake as the basic mechanism of hyperregulation in dilute media. Reduction of serum osmolalities following injection of ouabain into the hemolymph confirms previous reports on localization of the sodium pump in the basolateral parts of epithelial cells.     

Phosphate uptake behavior of natural phytoplankton during exposure to solar ultraviolet radiation in a shallow coastal lagoon

The effect of solar UV radiation (UVR: 290–400 nm) on the ³²P-phosphate uptake rates of natural phytoplankton from a southern Atlantic Ocean coastal lagoon was studied during two consecutive summers at one station located in the marine-influenced area. Due to the shallowness of this lagoon and also to the generally high UV water transparency in this area, phytoplankton are exposed to high UV irradiances. The ³²P-phosphate uptake rates measured at several phosphate concentrations were inhibited up to 59.2% by UVR, although uptake stimulation was also observed in four of nine experiments (up to 28%). The effect of UVR on the apparent maximum velocity of ³²P-phosphate uptake (V _uptake) ranged from an inhibition of 49% to a stimulation of 31%. Although the highest inhibition values were associated with the maximum registered incident UV irradiance, a significant correlation between these two parameters was not observed. Changes in microalgal community structure were not related to the observed UV effect; however, a significant relationship was found between the inhibition of ³²P-phosphate uptake rates and V _uptake used as a proxy for phosphate deficiency. This relationship suggests that the phytoplankton phosphorus nutritional status modulates their sensitivity to UV exposure. Overall, our results suggest that solar UVR has the potential to affect phosphorus cycling.Communicated by O. Kinne, Oldendorf/Luhe     

Petroleum hydrocarbons in the marine bivalve Venus verrucosa: accumulation and cellular responses

Cellular and subcellular responses of the marine burrowing bivalve Venus verrucosa collected from the north-eastern coastline of Malta from January to June 1985, after exposure to petroleum hydrocarbons (PHC) were investigated. After long-term exposure to 100 gl^-1 of water-accomodated fractions (WAF) of crude oil, PHC were found to accumulate most rapidly in the digestive gland and then in the gills, with saturation levels being reached within 100 d of exposure in both cases. PHC accumulation, both in the mantle and muscle tissues, was more gradual and consistent throughout the whole exposure period. After 150 d of exposure, the digestive cells of the digestive gland were significantly reduced in height (atrophy) and exhibited reduced lysosomal membrane stability. After 144 h of exposure to higher concentration of PHC (820 and 420 gl^-1), several cytological effects were recorded, including an increase in cell volume and activity of gill mucocytes as well as in the number of haemocytes in gill blood sinuses. There was also evidence of damage to the epithelial lining of the foot, stomach and style sac and marked atrophy of the digestive cells of the digestive gland. The significance of such responses is discussed.     

Uptake,binding and clearance of divalent cadmium in Glycera dibranchiata (Annelida: Polychaeta)

The bloodworm Glycera dibranchiata Ehlers, 1968 accumulates cadmium through the general body surface and the intestine. Absorption through the gut accounts for cadmium which rapidly binds to coelomic proteins. Intracoelomic injection of ¹⁰⁹Cd demonstrates that cadmium binds readily to hemoglobin and other proteins. The degree of cadmium binding is pH-dependent. The apparent pK of binding sites in body wall and musculature homogenates is 5.39. Cadmium ions injected into the coelom at 7 g g^-1 tissue increase proline incorporation rates into the positively charged hemoglobin (cathode fraction) by 15-fold in 3 days. A 3-fold increase of proline incorporation was observed in the anode hemoglobin fraction over the same time period. Radioactivity in various protein fractions decreases at different rates after injection of ¹⁰⁹Cd. Comparisons between the function of mammalian metallothionein and the coelomic fluid proteins of G. dibranchiata as a detoxification mechanism are discussed.     

Nitrogen uptake and growth of the germlings and mature thalli of Fucus distichus

Fucus distichus L. was collected near Vancouver, Canada, in late fall and early winter, 1981. The effects of the forms of nitrogen (nitrate, ammonium or urea) and periodic exposure to air on growth, rhizoid development and nitrogen uptake in germlings was investigated. Gamete release, fertilization, germination and germling growth had no requirement for a specific form of nitrogen. Periodic exposure to air increased secondary rhizoid development twofold. Nitrate and ammonium uptake rates of the germlings were higher than for the mature thalli (20 to 40 times for nitrate and 8 times for ammonium), while the halfsaturation constant (K _s) values for nitrate were similar (1 to 5 M). The germlings showed saturable uptake kinetics but the mature thalli did not. When germlings were exposed to air it caused a 70% decrease in nitrate uptake, but not change in ammonium uptake. Ammonium uptake in the mature thalli was proportional to the ambient ammonium concentration. Nitrate uptake in the mature thalli appeared to follow saturation kinetics at low nitrate concentrations, but showed a non-saturable component at concentrations greater than 10 M. Presence of ammonium inhibited nitrate uptake by the mature plants but not by the germlings.