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J Appl Phycol (2006) 18:135143 DOI 10.

1007/s10811-006-9084-0

ORIGINAL ARTICLE

Adsorption of radionuclides (134 Cs, 85 Sr, 226 Ra, 241 Am) by extracted biomasses of cyanobacteria (Nostoc carneum, N. insulare, Oscillatoria geminata and Spirulina laxissima) and Phaeophyceae (Laminaria digitata and L. japonica; waste products from alginate production) at different pH
Peter Pohl Wolfgang Schimmack

Received: 9 September 2005 / Revised and Accepted: 8 March 2006 C Springer Science + Business Media B.V. 2006

Abstract The extracted biomasses of four cyanobacteria (Nostoc carneum, Nostoc insulare, Oscillatoria geminata, and Spirulina laxissima) grown in axenic mass cultures, and of four samples of Laminaria obtained from different locations (L. digitata I and II, France; L. japonica I and II, China; all waste products from alginate production) were tested for their ability to adsorb four radionuclides (134 Cs, 85 Sr, 226 Ra, and 241 Am) under different pH regimes. In addition, two of the cyanobacterial biomasses (N. carneum and O. geminata) and the four Laminaria biomasses were phosphorylated before being tested as radionuclide adsorbers. The non-phosphorylated cyanobacterial biomasses showed very low adsorption of 134 Cs but substantially higher removal of 85 Sr and 226 Ra, which increased with increasing pH. 241 Am was almost completely removed from the solution at low pH, but less at higher pH. After phosphorylation, removal of 134 Cs, 85 Sr and 226 Ra by the cyanobacterial biomasses was improved, particularly at lower pH, but there was almost no adsorption of 241 Am. The non-phosphorylated

Laminaria biomasses showed good removal of 134 Cs and very good adsorption of 85 Sr and 226 Ra. Removal of 241 Am was high at low pH but decreased with increasing pH. After phosphorylation, adsorption of 134 Cs by Laminaria samples was slightly improved; removal of 85 Sr and 226 Ra was increased at low pH with a tendency towards decrease in adsorption with increasing pH; but almost no 241 Am was adsorbed. The origin of the cyanobacterial and Laminaria materials appeared to have little effect on the adsorption of the radionuclides. Keywords Algae, Drinking water Radioactivity removal Radionuclide adsorption Waste water treatment

Introduction Many authors have reported that extracted (non-living) biomasses of algae, bacteria and fungi can be used as adsorbers of metal ions from polluted water (Fehrmann & Pohl, 1993; Holan et al., 1993; Wilde & Benemann, 1993; Sandau et al., 1996; Fourest & Volesky, 1997; Pan et al., 2000; Radway et al., 2001; Nigro et al., 2002; De Philippis et al., 2003). The potential practical importance of such biological materials for controlling the discharge of radioactive efuents was already recognized in the 1950s and 1960s (Pickering & Lucas, 1962). Compared to conventional treatment methods,
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P. Pohl ( ) Universit t Kiel, Pharmazeutisches Institut, Abteilung a Pharmazeutische Biologie, Gutenbergstr. 76-78, D-24118 Kiel, Germany e-mail: pepohl@web.de W. Schimmack GSF-National Research Center for Environment and Health, Institute of Radiation Protection, Ingolstaedter Landstr. 1, D-85764 Neuherberg, Germany

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biomass based systems are likely to be more acceptable in terms of both cost effectiveness and efciency (Sar et al., 2004). However, while research on bioremoval of heavy metals has grown immensely in the last decades, there have been few studies on the biosorptive uptake of radionuclides (RN) (Sar et al., 2004). Research with RN has focussed mainly on the removal of uranium and radium from mining waste water due to its importance for the disposal of the uranium mining and milling tailings (for a review see Kalin et al., 2004). However, there are other RN that are important, especially in relation to the nuclear industry. The Chernobyl accident in 1986 showed that the long-lived RNs 90 Sr and 137 Cs can present a longterm problem in terms of contamination of drinking water. The initial content of 137 Cs in the water of the Pripyat river was a factor of 2.5 higher than the UKs Generalized Derived Limits (GDL) for drinking water (100 Bq L1 ) (Smith et al., 2001). Although the concentrations of most RN declined rapidly to insignicant levels during the weeks following the accident, the 90 Sr levels still exceeded the GDL (50 Bq L1 ) in many cases as late as 1991. Reduction of the RN concentration in drinking water below the limits set by regulating authorities may be of crucial importance after a nuclear accident. Smith et al. (2001) suggest that the most effective way to reduce the doses resulting from consumption of contaminated drinking water is to remove the contaminants at the water supply and treatment stage. For this reason, we tested the suitability of biomass based systems for the adsorption of four radionuclides from water: 134 Cs and 85 Sr, used as short-lived substituents of 137 Cs and 90 Sr, the most important anthropogenic radionuclides resulting from nuclear accidents; 226 Ra as a prominent natural radionuclide (member of the 238 U decay series); and 241 Am as a representative of the transuranic elements. Moderate concentrations of these radionuclides were used in order to simulate the type of situations that arose after the Chernobyl accident, i.e. contamination of water bodies used for drinking water at a level signicantly higher than permitted for consumption, but much lower than in nuclear waste liquids. Our previous studies on the removal of heavy metals from contaminated water showed that extracted biomasses of cyanobacteria, and in particular of Phaeophyceae, have high adsorption capacities (Fehrmann & Pohl, 1993; Winter et al., 1994). In particular, we
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found that waste materials from alginate production from Laminaria species (Phaeophyceae) can be puried and used as potent heavy metal adsorbers (Pohl et al., 2000). Such waste products are available in large quantities from alginate manufacture in many countries including France, China, Canada, and Norway. We therefore decided to test these materials for the ability to adsorb RN. Both our studies and those of other authors have shown that the pH of water samples is very important for heavy metal adsorption, with an optimum between pH 5 and pH 9 (Pan et al., 2000; Radway et al., 2001). At low pH (<3) heavy metal adsorption decreases signicantly (Fehrmann & Pohl, 1993; Kratochvil & Volesky, 1998; Pan et al., 2000). Thus the experiments with radionuclides were carried out at four different levels of pH ranging from about pH 3 to about pH 9. Furthermore, results obtained by Klimmek et al. (2001) suggest that the adsorbing capacity of algal biomasses might be increased by phosphorylation, as was shown for the cyanobacterium Lyngbya taylorii. Thus we also tested the RN adsorbance capacity of extracted biomasses of several species after phosphorylation.

Materials and methods Cyanobacteria Four cyanobacteria samples were tested: Nostoc carneum Vaucher a gift from the Forschungsinstitut Senkenberg, Frankfurt a.M., Germany; and Nostoc insulare Borzi 54.79, Oscillatoria geminata Meneghini B 1459-8, and Spirulina laxissima G.S. West B 256.80 all from the Collection of Algal Cultures, G ttingen, Germany. The microalgae were grown at o 22 C under axenic conditions and continuous aeration in asks containing 8 L of an inorganic medium consisting of 90% demineralised water and 10% seawater with added phosphate, nitrate and trace elements (DS-medium; Pohl et al., 1987). The nitrogen-xing organisms Nostoc carneum and N. insulare were grown without additional nitrogen, and the non-nitrogenxers Oscillatoria geminata and Spirulina laxissima were grown with KNO3 as nitrogen source. The cultures were illuminated with uorescent tubes (Philips TL 65W/25; white) at 25 mol photons s1 m2 . After reaching the stationary growth phase, the cyanobacteria were harvested by centrifugation, washed 5 times with

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demineralised water and freeze-dried. The biomass was extracted exhaustively with methanol (80%) in a Soxhlet apparatus, washed 5 times with demineralised water and dried at 80 . The resultant mass was ground in a mortar and sieved to a particle size of 0.50.8 mm. Phaeophyceae Four Laminaria samples were obtained as waste products from alginate production. Two samples of Laminaria digitata (L.) Lamour were obtained from Degussa Texturant Systems, Lannilis, France, as moist and crushed waste products. The original algae from which these samples were obtained were collected from two different locations on the Atlantic coast of France. Two samples of Laminaria japonica Lamour were obtained from Longtai Chemical Industrial Co., Tianjin, China, also as waste products but in dried and powdered form (kelp residue). All four samples were extracted three times with demineralised water at 80 C under continuous stirring. The resultant puried materials were dried at 80 C, ground, and sieved to a particle size of 0.50.8 mm. Phosphorylation Aliquots of each of the puried Laminaria materials and of two of the cyanobacteria samples (O. geminata and N. carneum) were heated to 170 C with urea and phosphoric acid according to the method described by Klimmek et al. (2001). After cooling, the materials were washed thoroughly with demineralised water, dried at 80 C and sieved to a particle size of 0.5 0.8 mm. An additional sample of L. japonica II was sieved to a particle size of 11.5 mm for comparison and to demonstrate the importance of having a uniform particle size. Adsorption of radionuclides 10 mg dry alga mass was suspended in 9.9 mL of distilled water in a centrifuge bottle and roughly adjusted to a given pH (about 3, 5, 7 or 9) by adding HCl or NaOH; 0.1 mL of a slightly acidic stock solution with the radionuclides was then added and the pH adjusted again, and the nal pH value measured with a microelectrode. The 0.1 mL stock solution contained 15,700 Bq 85 Sr (purchased from PerkinElmer Life Sciences, Boston, USA), 420 Bq 134 Cs (RISO

National Laboratories, Ros-kilde, Denmark), 510 Bq 226 Ra (AEA Technology, Braunschweig, Germany), and 290 Bq 241 Am (PTB Braunschweig, Germany). The 226 Ra was carrier-free, the carriers for 85 Sr and 134 Cs were the corresponding stable elements, and the carrier for 241 Am was La(NO3 )3 . The total amount of elements and radionuclides in each centrifuge bottle was thus 1 ng Cs (134 Cs and stable Cs), 23 ng Sr (85 Sr and stable Sr), 14 ng226 Ra, and 2.3 ng 241 Am (with 101 ng La). The theoretical load of the adsorbing material was 0.1 g g1 biomass for Cs, 2.3 g g1 for Sr, 1.4 g g1 for 226 Ra, 0.23 g g1 for 241 Am and 10 g g1 for La. The suspension was shaken by rotation at approximately one rotation per second for 30 min; centrifuged at 4500 rpm for 10 min; and ltered (0.45 m, Sartorius). One ml of the ltrate was added to 34 mL distilled water for gamma-measurement (see below), and the nal pH measured in the remainder of the ltrate. If the difference between the initial pH value and the nal pH value was greater than 0.1, the experiment was repeated. The amount of RN activity sorbed to the algal biomass was determined from the difference between the RN activity added to the solution and the activity remaining in the solution after 30 min. The pH stability of the radionuclides was tested as follows. Control samples without algal material were prepared by diluting 0.1 mL of radionuclide stock solution in 9.9 mL distilled water in a centrifuge bottle and adjusting to pH values between 3 and 10. The samples were shaken, ltered, and centrifuged as described above, and the radionuclides in the ltrates measured. The results showed that there was no change in radionuclide content in the samples, thus within this pH range there was no sorption to the walls of the centrifuge bottles. No hydrolysis effects were observed for any of the RN for alkaline pH values due to their low concentration in the solution. Determination of the radionuclides The radionuclides were determined by direct gamma spectrometry using a high purity Ge-detector and a multichannel analyser. Line 604.7 keV was used for evaluation of 134 Cs, and the count rate corrected for sum coincidence. There were no net pulses (after background subtraction) without sample for 85 Sr (514.0 keV), 134 Cs, or 241 Am (59.5 keV). A net pulse of about 0.005 counts per second had to be taken into account for 226 Ra (186.2 keV). This meant that in general
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the experimental uncertainty of 226 Ra was between 10 and 20%, while that of the other RNs was <5%. All activities were decay-corrected for August 1, 2002. The adsorption was characterized by the radionuclide activity removed per g biomass as well as by the adsorption ratio, i.e. the adsorbed activity as a percentage of the radionuclide activity added to the solution. The concentration factors of the radionuclides, often found in the literature, can be obtained from the adsorption ratio by multiplying by 10, because the mass ratio of solution to algae was 1000.

Results The results of the adsorption experiments are shown in Tables 13. A total of 15 samples were tested, each at four different levels of pH. (The pH of individual samples varied somewhat as a result of the difculty of adjusting pH accurately). In each case, the adsorption is shown in terms of units of weight (g g1 ) and percentage of total RN added. The amounts added for each RN were different as they were based on the amount needed to obtain a useful signal. Thus the adsorption data for different radionuclides should be compared in terms of the percentage adsorption rather than the total radionuclide uptake per g biomass. For example, Nostoc carneum at pH 3.9 adsorbed 5.88 ng of Sr, or 0.588 g g1 biomass, and 2.29 ng of 241 Am, or 0.229 g g1 biomass (Table 1). Thus 25.6% of the total 23 ng of Sr added (85 Sr and stable Sr) was adsorbed, and 99.4% of the total 2.3 ng of 241 Am added. Although the amount of 241 Am adsorbed per g biomass was less than half of that of Sr, the percentage adsorption of 241 Am was much higher. These adsorption data are only valid as long as the experiments are carried out within the linear adsorption range of the materials. This was ensured by the low RN quantities used (see below). Table 1 shows the results of adsorption by the cyanobacterial biomasses. All four nonphosphorylated samples showed very low adsorption of 134 Cs, less than 3% at low pH rising to only 7 to 13% at high pH. Three of the four biomasses adsorbed less than 10% of 85 Sr at low pH, but adsorption increased at higher pH, reaching values of more than 90% (Oscillatoria and Spirulina). Removal of 226 Ra was similar to that of 85 Sr, being fairly low at pH 34 but increasing signicantly to as high as 96% at pH
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values above 5 (Oscillatoria). All samples showed high adsorption of 241 Am (70 to 99%) at low pH, and reduced adsorption at higher pH, down to almost zero in the case of S. laxissima Adsorption of 134 Cs by N. carneum and O. geminata increased markedly after phosphorylation, to almost 80% at high pH. Removal of 85 Sr also increased markedly, especially at lower pH. However, at the highest pH, removal of 85 Sr by the phosphorylated Oscillatoria sample was lower than removal by the non-phosphorylated sample. Adsorption of 226 Ra was also increased at lower pH (up to 98.6%) and decreased at higher pH. The phosphorylated samples adsorbed almost no 241 Am. Table 2 shows the results of adsorption by the nonphosphorylated Laminaria samples. All four Laminaria samples adsorbed considerably more 134 Cs than did the cyanobacteria biomasses, removing between 35 and 50% at low pH, and up to 80% at higher pH. Adsorption of 85 Sr by the Laminaria species was also slightly higher overall than for the cyanobacteria biomasses, between 30 and 70% at lower pH increasing to nearly 100% at higher pH. Removal of 226 Ra was high (7599%) for all four biomass samples and at all pH values, with no reduction at higher pH values as displayed by the cyanobacteria biomasses. All four Laminaria samples adsorbed high amounts of 241 Am at low pH with a steady decrease with increasing pH, but the decrease was somewhat less marked than for the cyanobacteria biomasses. Table 3 shows the results of adsorption by the phosphorylated Laminaria samples including one sample with larger particle size. Overall, removal of 134 Cs was slightly improved compared to the non-phosphorylated samples. Removal of 85 Sr was clearly enhanced with values of up to 97% at low pH, but with a tendency towards decrease in adsorption with increasing pH, with the exception of L. digitata I, which showed high 85 Sr adsorption at all pH values. The removal of 226 Ra was similar to that of 85 Sr with values above 90% at low pH and, in contrast to the non-phosphorylated samples, a marked decrease in absorption with increasing pH. As with the phosphorylated cyanobacterial biomasses, the phosphorylated Laminaria samples adsorbed almost no 241 Am. The results for the Laminaria japonica II sample with larger size particles (1.01.5 mm) (Table 3) show clearly that the larger particles were less effective in removing the radionuclides than the smaller particles, particularly for 85 Sr and 226 Ra.

J Appl Phycol (2006) 18:135143 Table 1 Adsorption of radionuclides (RN) by extracted algal materials from cyanobacterianon-phosphorylated and phosphorylated (g RN per g biomass and % of added RN)a
134

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Cs % g g

85 1

Sr %

226

Ra %

241

Am %

pH Nostoc carneum 3.9 5.1 6.6 9.1 Nostoc insulare 3.9 5.4 6.8 9.6 Oscillatoria geminata 3.2 5.7 6.9 9.9 Spirulina laxissima 3.3 5.5 7.2 10.1 Nostoc carneum (phosphorylated) 3.5 5.7 7.2 9.5 Oscillatoria geminata (phosphorylated) 3.5 5.1 7.5 9.6
a b

g g

g g

g g1 0.229 0.091 0.011 0.052 0.197 0.167 0.059 0.023 0.222 0.203 0.190 0.102 0.160 0.155 0.151 0.004 0.000 0.005 0.000 0.003 0.000 0.009 0.000 0.007

0.003 0.002 0.003 0.007 0.000 0.002 0.003 0.008 0.000 0.003 0.004 0.011 0.003 0.004 0.006 0.013 0.053 0.076 0.079 0.079 0.034 0.061 0.075 0.078

2.7 2.3 3.1 7.3 <DLb 2.0 2.9 7.9 <DL 2.9 3.7 10.9 2.6 4.0 6.2 13.1 53.3 75.8 79.0 79.3 33.9 61.4 75.4 78.0

0.588 1.009 1.240 1.660 0.197 0.820 1.186 1.721 0.214 1.471 1.624 2.079 0.108 1.408 1.842 2.127 2.250 2.203 2.064 1.783 1.890 1.967 1.912 1.732

25.6 43.9 53.9 72.2 8.6 35.7 51.6 74.8 9.3 64.0 70.6 90.4 4.7 61.2 80.1 92.5 97.8 95.8 89.7 77.6 82.2 85.5 83.1 75.3

0.756 1.165 1.297 1.284 0.217 0.875 1.163 1.229 0.378 1.327 1.346 1.345 0.123 1.274 1.313 1.245 1.381 1.290 1.124 0.818 1.227 1.207 0.893 0.787

54.0 83.2 92.7 91.7 15.5 62.5 83.1 87.8 27.0 94.8 96.2 96.1 8.8 90.9 93.8 88.9 98.6 92.1 80.3 58.5 87.6 86.2 63.8 56.2

99.4 39.8 4.6 22.7 85.7 72.7 25.4 10.0 96.7 88.1 82.5 44.3 69.4 67.2 65.6 1.5 <DL 2.4 <DL 1.2 <DL 4.1 <DL 3.1

The particle size was 0.5-0.8 mm DL = detection limit, about 0.5%

Discussion and conclusions Non-phosphorylated and phosphorylated biomasses of several cyanobacteria and algae were tested for their ability to eliminate four relevant radionuclides (RN) from aqueous solutions at different pH values. Overall, the results show that RN at the concentrations likely to be found in contaminated water supplies could be removed effectively, but the extent of removal

varied with pH, the specic type of RN, and whether or not the biomasses were phosphorylated. 85 Sr and 226 Ra were well adsorbed between pH 5 and 9 by all non-phosphorylated materials, but somewhat less well by most of these materials at low pH, thus they resembled the heavy metals (Cd, Cr, Cu, Hg, Ni, Pb and Zn) which we had investigated previously (Pohl et al., 2000). 134 Cs was adsorbed well by the nonphosphorylated biomasses of the Laminaria species,
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140 Table 2 Adsorption of radionuclides (RN) by extracted algal materials from Phaeophyceae non-phosphorylated (g RN per g biomass and % of added RN)a

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Cs % g g1

85

Sr %

226

Ra %

241

Am %

pH

g g1

g g1

g g1

Laminaria digitata I 3.5 5.7 6.9 10.6 0.036 0.056 0.057 0.066 35.5 56.2 56.9 65.6 0.670 1.383 1.601 1.995 29.1 60.2 69.6 86.7 0.968 1.205 1.247 1.304 69.2 86.1 89.1 93.1 0.223 0.204 0.195 0.111 97.0 88.8 84.9 48.3

Laminaria digitata II 3.4 4.9 6.5 9.7 0.04544.6 0.06060.0 0.06260.6 0.07271.7 0.904 1.443 1.833 2.181 39.3 62.8 79.7 94.8 1.052 1.232 1.316 1.315 75.1 88.0 94.0 93.9 0.227 0.219 0.199 0.107 98.5 95.4 86.7 46.6

Laminaria japonica I 3.3 5.6 7.1 10.6 0.049 0.058 0.069 0.082 49.3 57.7 69.1 82.0 1.583 68.8 2.090 90.9 2.215 96.3 2.266 98.5 1.339 95.7 1.348 96.3 1.386 99.0 1.378 98.4 0.226 98.2 0.181 78.6 0.155 67.5 0.038 16.3

Laminaria japonica II 3.3 5.3 7.1


a

0.042 0.059 0.063 0.079

42.4 59.5 63.3 78.9

1.440 2.018 2.111 2.184

62.6 87.7 91.8 95.0

1.290 1.308 1.363 1.334

92.1 93.4 97.3 95.3

0.216 0.199 0.182 0.105

93.9 86.7 79.3 45.6

The particle size was 0.50.8 mm

10.3

especially at higher pH, but not by non-phosphorylated biomasses of cyanobacteria. Following phosphorylation, all biomasses showed good and even excellent adsorption of all three of these RN. The results with 241 Am were very different with almost no adsorption by phosphorylated species, and good adsorption by non-phosphorylated materials at low pH decreasing at higher pH, the reverse of the situation found with the other three RN. The results obtained among the different cyanobacterial species were similar, as were those obtained among the different Laminaria species, indicating that the origin of samples is of minor importance. Larger particles were less effective in removing the radionuclides than smaller particles, particularly for 85 Sr and 226 Ra, which was expected because they have a smaller surface area per unit of weight. This study was carried out mainly with a view to investigating potential methods for treating the type of drinking water contamination likely to be found in an area affected by a nuclear accident, as happened in the region of Chernobyl after 1986 (Smith et al., 2001).
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Thus moderate concentrations of RN were used; much lower than those present in industrial nuclear waste waters. There is a further potential use of methods for removing RN at moderate concentrations. Conventional methods for treatment of waste water containing high concentrations of heavy metals, for example chemical precipitation combined with ltration, still result in aqueous solutions that contain contaminants in concentrations signicantly above the legal limits (Volesky, 1987). This is likely to be the case for nuclear waste waters as well. Thus it is likely that further decontamination of treated waste water will be needed. This can be achieved by membrane technologies, but the results reported here indicate that use of adsorbing materials is also a possibility. Moderate RN concentrations have also been used in investigations reported by others, for example investigations into the bioaccumulation of some RN by two freshwater algae by Adam and Garnier-Laplace (2003) and by Boisson et al. (1997). The radiation from high RN concentrations can alter the biomass structure. In the experiments reported here, the

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Table 3 Adsorption of radionuclides (RN) by extracted algal materials from Phaeophyceae phosphorylated (g RN per g biomass and% of added RN)a
134

Cs % g g

85 1

Sr %

226

Ra % g g

241 1

Am % <DLa <DL <DL 6.5 <DL <DL <DL <DL 2.4 3.7 <DL <DL <DL <DL <DL <DL <DL <DL <DL <DL

pH Laminaria digitata I 3.0 5.8 7.1 9.9 Laminaria digitata II 3.1 5.3 7.0 9.8 Laminaria japonica I 3.4 5.7 7.3 9.8 Laminaria japonica II 3.5 5.9 7.1 9.5 Laminaria japonica II (larger particles: 1.01.5 mm) 3.4 5.7 7.2 9.9
a b

g g

g g

0.042 0.082 0.085 0.088 0.063 0.079 0.082 0.082 0.030 0.072 0.075 0.076 0.032 0.068 0.071 0.074 0.029 0.064 0.069 0.072

42.2 82.2 84.6 88.2 62.8 78.7 81.5 82.1 30.1 71.6 74.8 75.7 31.8 68.5 71.2 73.7 28.9 64.2 69.3 72.3

2.119 2.248 2.242 2.242 2.107 2.167 1.978 1.596 2.242 2.104 2.137 1.756 2.117 1.623 1.489 1.418 1.232 1.149 1.043 0.917

92.1 97.7 97.5 97.5 91.6 94.2 86.0 69.4 97.5 91.5 92.9 76.3 92.1 70.6 64.8 61.6 53.5 49.9 45.3 39.9

1.356 1.342 1.316 1.243 1.273 1.282 1.128 0.685 1.353 1.240 1.231 0.842 1.282 0.894 0.794 0.528 0.712 0.638 0.432 0.267

96.9 95.9 94.0 88.8 91.0 91.6 80.6 48.9 96.7 88.6 87.9 60.1 91.6 63.9 56.7 37.7 50.9 45.6 30.8 19.1

0.000 0.000 0.000 0.015 0.000 0.000 0.000 0.000 0.005 0.009 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000

If not stated otherwise, the particle size was 0.50.8 mm DL = detection limit, about 0.5%

radiation dose of the RN was clearly below such levels. Thus, the biosorption of the RN can be considered in the same way as the biosorption of a nonradioactive metal species (Tsezos, 1990). Because only very small amounts of the radionuclides were added to the suspensions (0.12.3 g g1 biomass), it can be assumed that the adsorption isotherm was linear in all the adsorption experiments described. Our studies with heavy metals have shown that adsorbing algal materials can bind up to 110130 mg of heavy metal per g biomass, with linear adsorption up to ca. 100 mg of heavy metal per g biomass (data not yet published). Other authors

working with algae have reported comparable heavy metal adsorption rates (Klimmek et al., 2001). In the heavy metal studies, maximum adsorption was obtained after about 5 min, thus it was assumed that an adsorption time of 30 min would be sufcient for the RN experiments. The four cyanobacterial species used in the experiments were grown in the laboratory under controlled and comparable conditions. This method of mass cultivation has the advantage of yielding well-dened biomasses reproducibly. Highly puried cyanobacterial cell wall materials can be obtained after extraction of otherwise usable compounds like lipids,
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sterols, carotenoids and phycobiliproteins (Pohl, 1982; Kohlhase & Pohl, 1988; Volk, 1996). The cell surface consists predominantly of mureine, a peptido-glycan typical for prokaryotic organisms. The chemical structure of the surface may be responsible for the RN adsorption pattern. The hydroxy and N-acetyl groups in mureine presumably play an important role in RN adsorption. After phosphorylation, the hydroxy groups are converted either to phosphate groups, which certainly improve RN adsorption, or with participation of urea to carbaminic acid esters, which may bind RN via side valences. The Laminaria samples originated from macroscopic brown algae collected from coastal areas of France and China for the purpose of industrial alginate production. The waste product obtained after extraction of these compounds, also consists mainly of cell wall material contaminated with alginates, salts, proteins, pigments and others. These impurities had to be removed thoroughly (see Materials and Methods). The cell wall material of Phaeophyceae is mainly composed of cellulose microbrills, xylofuco-glycans, xylo-fuco-glycuronates and homofucans (Kloareg et al., 1986). These polysaccharides have OHand SH-groups as well as acidic functional groups such as carboxy and sulphate groups (Kuyucak & Volesky,1990; Fehrmann & Pohl, 1993; Kratochvil & Volesky, 1998). Especially the carboxy and sulphate groups will lead to strong RN adsorption. After phosphorylation, the hydroxy groups are converted to phosphate groups, and the carboxy groups with participation of NH3 from urea to uronic acid amides, which might also offer side valence bondings. Furthermore, it can be assumed that the sulphate groups were partly or totally transesteried to phosphate groups. The binding capacities of the various functional groups may also depend on the conguration of the polysaccharide polymer (Holan et al., 1993). As a result of the rather complex structure of the biomass surfaces, it is only possible to explain the differences in the adsorption of the RN in part. The non-phosphorylated cyanobacterial and Laminaria biomasses removed smaller amounts of the monovalent 134 Cs+ than of the divalent cations 85 Sr++ and 226 Ra++ . Poor adsorption of 134 Cs by marine algae and picoplankton, in comparison with adsorption by divalent radionuclides, has also been reported by Fisher (1985), Boisson et al. (1997), Heldal et al. (2001) and Adam and Garnier-Laplace (2003). This is likely to
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be because the equilibrium constants of complexes of divalent cations with carboxylic acids are larger by orders of magnitude than those of monovalent cations (Pohlmeier, 1999). According to the general rule for complex stability the higher the charge the greater the stability constantthe adsorption of the trivalent 241 Am should have been even greater than that of 85 Sr and 226 Ra. In practice, this was observed only at pH 3 (Tables 1 and 2). At pH 5, several biomasses showed a higher adsorption of 85 Sr and 226 Ra than of 241 Am, and all at pH 7 and 9, with the effect especially marked for cyanobacteria. Boisson et al. (1997), while investigating the temperature dependence of the adsorption of 11 radionuclides by the marine alga Fucus vesiculosus (L.) at pH 7.8, also found that the adsorption of 241 Am was smaller than that of divalent cations like 65 Zn. The causes of this effect are as yet unknown. After phosphorylation, there was a marked improvement in adsorption of 134 Cs by all biomasses, as well as an improvement in adsorption of 85 Sr and 226 Ra, particularly at low pH. This is probably the result of transformation of the hydroxy groups present in the non-phosphorylated materials into phosphate groups by the phosphorylation process. Most strikingly, however, adsorption of 241 Am virtually ceased following phosphorylation. Hence phosphorylation will not always lead to an improvement, it depends on the adsorbant. Clearly, the original binding sites for 241 Am in the non-phosphorylated materials must have been blocked or destroyed by the phosphorylation process, and the acidic phosphate and sulphate groups or the carbaminic acid esters and uronic acid amides present on the surface of the phosphorylated materials were not able to bind 241 Am. The following conclusions can be drawn. Puried (extracted) material from the four Laminaria species investigated can be used as a potent adsorber of 85 Sr, 226 Ra, and 241 Am, and to a somewhat lesser extent of 134 Cs, in the range pH 3 to pH 10. Absorption of 85 Sr, 226 Ra, and 134 Cs was further improved at lower pH after phosphorylation. The cyanobacterial biomasses were also medium to good adsorbers of 85 Sr, 226 Ra, particularly at higher pH, and of 241 Am at low pH. Phosphorylation markedly improved adsorption of 134 Cs and 85 Sr, and to a lesser extent 226 Ra. Adsorption was pH sensitive. None of the materials adsorbed 241 Am after phosphorylation. The origin of the Laminaria materials and the different nitrogen requirements of the cyanobacteria

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appeared to be of minor importance for the adsorption of the RN. The ndings suggest a possibility for practical application in purication of contaminated water and secondary purication of partially treated nuclear waste. This could offer an opportunity for turning cheap waste materials derived from renewable resources into valuable new products. Acknowledgements We thank Frauke Rodde and Wolfgang Schultz for their excellent technical assistance, and Dr A. Beatrice Murray for editing the English text. References
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