Die Massenentwicklung der Geißelalge Uroglena americana (Alk.) Lemm in Trinkwassertalsperren Thüringens und ihre Bekämpfung

A mass development of Uroglena americana with cellular densities of up to 13.5 · 10⁶ and 86.10⁶ cells/l, was observed in two drinking-water reservoirs. In the first case the development remained confined to the upper 20 m, in the second case there occurred a gradual shift of the maximum to deeper zones from May to July with decreasing cellular densities down to a depth of 35 m. Laboratory experiments for the control of the algae development were performed in order to protect the drinking-water resources from potential impairments of odour and taste. Samples of the storage-reservoir water with 20.10⁶ cells/l of Uroglena were concentrated up to 1.25 … 12.5 mg/l Ca(OH)₂ (pH-values of 7.2 … 9.6) by addition of calcium hydroxide and observed for 39 h. First impairment of the cells of Uroglena is detectable at 5 mg/l Ca(OH)₂ after 15 h, an immediate effect can be observed at 7.5 mg/l Ca(OH)₂ together with increased sedimentation, also Gymnodinium and Cryptomonas having been damaged. Thereupon a successful control of Uroglena by liming up to pH-values above 8.0 is possible. The application of lime to outdoor areas was not necessary, in the final analysis, since the drinking-water quality could be secured by the optimum choice of the intake depth of raw water with a maximum of 4.10³ to 77.10³ cells/l.     

Interaction between Scenedesmus obliquus and the Heavy Metals Copper and Lead

The effects of the addition of copper (in copper sulphate and copper chloride) and lead (in lead nitrate and chloride) on Scenedesmus obliquus were studied. The laboratory studies have shown that Sc. obliquus is able to accumulate copper and lead to some extend depending on the concentration of the metal and on the time of contact of algae with the metal. Copper in the concentration of 1 mg/l caused the decrease of the total protein and the growth of the population as well as the destruction of histons, peptids chains and vacuolization of cells. Such drastic effects were observed in the higher concentration of lead (100 mg Pb/l).     

On the ecology of Azolla filiculoides Lam. in Damietta district, Egypt

Azolla filiculoides Lam. is a tiny aquatic fern growing as free floating mats covering the stagnant water of the irrigation and drainage canals of Damietta district, Egypt. Ecological characteristics and distribution of the fern were investigated and results were discussed.Vegetation analysis of Azolla indicate that the most common associated species with Azolla are: Phragmites australis [presence (P) = 93.3%], Echinochloa stagnina (P = 70%), Eichhornia crassipes (P = 57%), Pluchea dioscoridis (P = 56.7%), Persicaria salicifolia (P = 26.7%), Phylla nodiflora (P = 13.3%) and Ludwigia stolonifera (P = 10%). Submerged species, e.g. Ceratophyllum spp. and Potamogeton spp. are completely absent in the stands dominated with solid mats of Azolla. This could be explained by that these mats prevent light penetration and oxygen which are required to submerged plants which in turn are decayed.The pH of the water where Azolla was recorded ranged from 7.1 to 9.0 (mean = 7.8), total dissolved salts ranged from 280 to 2600 mg/l (mean = 880 mg/l). The total phosphorus showed the range from 0.01 to 0.82 mg/l (mean = 0.22 mg/l). The ammonia-N is relatively high, varying from 2 to 18 mg/l (mean = 7.0 mg/l), however, the nitrate-N varied from 2.1 to 54.1 mg/l with a mean value of 20.5 mg/l.The fresh biomass of Azolla ranged from 114 to 4280 g/m² in irrigation canals and from 506 to 2760 g/m² in drainage canals. The dry biomass varied from 16 to 23 g/m² in irrigation canals. The range from 26 to 320 g/m² was obtained for drainage canals.     

Biocides in Relation to Water Pollution. Part 2: Bioassay Studies of Few Biocides to a Fresh Water Fish,Channa gachua.

The present paper deals with the determination of LT_m values of zolone, rogor, sumithion and abate for Channa gachua by conducting static bioassay experiments. The TL_m values for zolone came out to be 0.08275 mg/l, 0.08225 mg/l, 0.0815 mg/l and 0.0810 mg/l; for rogor 5.155 mg/l; 4.955 mg/l, 4.65 mg/l and 4.475 mg/l; for sumithion 12.60 mg/l, 12.50 mg/l, 12.30 mg/l and 12.20 mg/l and for abate 217.25 mg/l, 225.575 mg/l, 221.575 mg/l and 217.25 mg/l for 24, 48, 72 and 96 hours, respectively. Relative susceptibility indicates that this fish is not susceptible to zolone and less to rogor, sumithion and abate, respectively. Present studies also show that increased temperature and pH increase the toxicity of all the biocides, but increased hardness of water decreases the toxicity of biocide solutions. The safe concentrations for zolone, rogor, sumithion and abate came out to be 0.02437 mg/l, 1.3733 mg/l, 3.6907 mg/l and 66.678 mg/l, respectively.     

Lethal and estrogenic effects of 4-nonylphenol in the cockle Cerastoderma glaucum

The lethal and sublethal effects of the xenoestrogen 4-nonylphenol (NP) were evaluated in the cockle Cerastoderma glaucum. In a 96-h lethality test, bivalves were exposed to 0, 0+ acetone, 0.19, 0.38, 0.75, 1.5 and 3.0 mg NP/l. The 96-h LC₅₀ value was 0.3 mg NP/l. No mortality was observed at 0.1 mg NP/l. The potential estrogenicity of NP was studied in both sexually undifferentiated (resting phase) and differentiated (pre-spawning phase) cockles, exposed for 7 and 14 days to 0, 0+ acetone, 0.0125, 0.025, 0.05, and 0.1 mg NP/l. Vitellogenin (Vg)-like protein levels were determined in both haemolymph and digestive gland by the alkali-labile phosphate (ALP) assay. In the resting phase, exposure for 7 days to 0.1 mg NP/l resulted in significant increases in ALP in both haemolymph and digestive gland, compared with controls. A significant increase was also observed in digestive gland of animals exposed to 0.0125 mg NP/l-exposed animals. After 14 days of exposure, haemolymph ALP levels were significantly increased in exposed animals at all NP concentrations tested, whereas no difference was recorded in digestive gland. In the pre-spawning phase, exposure for 7 days to NP significantly increased ALP levels in haemolymph from males exposed at all NP concentrations tested, whereas no significant variations were found in haemolymph from females. NP (0.05 and 0.1 mg/l) was also shown to increase ALP concentrations significantly in digestive gland of males, but not in those of females. Likewise, after 14 days’ exposure, ALP levels significantly increased in haemolymph from males only at 0.1 mg NP/l. Conversely, NP caused significant increases in ALP levels in digestive gland from both males (at all NP concentrations tested) and females (at 0.025 and 0.1 mg NP/l). These results demonstrate that NP induces Vg synthesis in C. glaucum. Interestingly, males were more responsive to NP than females.     

Immunotoxicity of the xenoestrogen 4-nonylphenol to the cockle Cerastoderma glaucum

The in vivo effects of 4-nonylphenol (NP) on functional responses of haemocytes from the cockle Cerastoderma glaucum were investigated after 7 days exposure to sublethal NP concentrations (0, 0+acetone, 0.0125, 0.025, 0.05 and 0.1 mg/l NP). Haemocytes from both controls and exposed cockles were collected, and the effects of NP on total haemocyte count (THC) and volume of circulating cells, intracellular superoxide anion () levels, acid phosphatase and lysozyme-like activities in both haemocyte lysate (HL) and cell-free haemolymph (CFH) were evaluated. Exposure of cockles to 0.1 mg/l NP significantly increased THC (p < 0.05) with respect to controls. Analysis of haemocyte size frequency distribution showed that the haemocyte fraction of about 7–8 μm in diameter and 250 femtolitres in volume increased markedly in cockles exposed to the highest NP concentration tested. Apoptosis resulting in cell volume reduction in NP-exposed animals cannot be excluded. No statistically significant variation in intracellular levels was observed. Conversely, significant increases (p < 0.05) in acid phosphatase activity were observed in CFH from 0.05 and 0.1 mg/l NP-exposed animals; no significant differences in enzyme activity were recorded in HL. Lysozyme-like activity also increased significantly in CFH from cockles exposed to 0.05 mg/l NP (p < 0.05) and 0.1 mg/l NP (p < 0.001). Instead, lysozyme-like activity decreased significantly (p < 0.05) in the HL of animals exposed to 0.05 mg/l NP. Our results suggest that NP induces variations in the functional responses of haemocytes of C. glaucum, mainly by reducing cell membrane stability and promoting cell degranulation.     

Einsatz einer konstruierten Mischkultur zur Verwertung von Mischsubstraten mit inhibierend wirkenden Substanzen

By way of introduction the general fundamentals and kinetic setups of the competitive and non-competitive as well as substrate inhibition of cultures of microorganisms are stated. Investigated on a laboratory scale is the degradation of liquid pig slurry in a 600-1 stirring fermenter with discontinuous addition (pO₂≧=30 = saturation) of methanol as external substrate by a yeast mixed culture of Metschnikowia and Pichia membranifaceans to utilize the steam-volatile fatty acids (12 g/l) and of Acetobacter methanolicus to utilize methanol. A stable fermentation with stable residual concentrations of 235 mg/l fatty acids, 142 mg/l mH₄-N an 100 … 300 mg/l methanol for aperiod of 2400 h can be achieved.     

Acute toxicity of cadmium to Channa punctatus (BLOCH)

Effects of acute cadmium poisoning on survival, its residual values and histopathology in certain organs of a freshwater airbreathing fish, Channa punctatus (BLOCH ) were investigated. The threshold concentration, MATC and LC₅₀ values obtained from 96 h static bioassay, revealed that Channa is more susceptible to cadmium ions at higher temperature. The atomic absorption spectrophotometric determination of cadmium residues differed significantly in organs of specimens having survived and died after 96 h of exposure. The gill accumulate the highest amount of cadmium, the liver accumulated a slightly smaller amount than the gill, while the kidney accumulated the least. The histopathological lesions subjected to sublethal (5.2 mg/l Cd) and lethal (8.4 mg/l Cd) concentrations of cadmium were: detachment and rupture of lamellar epithelium, collapse of pillar cells and hypertrophy in mucus producing gland cells in the gill; vacuolization and coagulative necrosis in hepatic cells of the liver; and expansion, necrosis and accumulation of cellular debris in renal tubules of the kidney. The probable causes for death of fish due to cadmium ions have been discussed.     

Limnology of an equatorial high mountain lake — Lago San Pablo, Ecuador: The significance of deep diurnal mixing for lake productivity

Equatorial high mountain lakes are a special type of lake occurring mainly in the South American Andes as well as in Central Africa and Asia. They occur at altitudes of a few thousand meters above sea level and are cold-water lakes (<20 °C). Relatively little is known about them. A long-term limnological study was therefore undertaken at Lake Lago San Pablo, Ecuador to analyze the basic limnological processes of this lake, which has a tendency for eutrophication. Lago San Pablo is spread over an area of 668 hectares, has a maximum depth of 35 m, and is located 2660 m above sea level. Its thermal stratification is a monomictic one, with only 1–2 °C difference between the epi- and hypolimnion; overturn is achieved by strong winds during the dry summer period. The stratification phase is characterized by an oxygen deficit in the lower part of the hypolimnion. Besides, strong convective currents occur due to nocturnal cooling, and partial lake mixing was observed during the nocturnal period. This type of lake mixing is called atelomixis, which is characterized by the partial mixing of isolated layers (difference in temperature or ionic content) during stratification. The nutrient level of the lake is quite high: mean P_total concentration = 0.22 mg/l, mean N_total = 1.05 mg/l, soluble reactive phosphorus (SRP) > 0.01 mg/l, and soluble inorganic nitrogen > 0.2 mg/l. Nitrogen and phosphorus are available in the epilimnion all year round (N_{sol. inorg·.} = 0.3 to 1.7 mg/l N, SRP = 0.04 to 0.63 mg/l P). The N/P ratio is sometimes > 14, sometimes < 10, indicating a variability of the limiting nutrient factor. Considering the nutrient level, the phytoplankton biomass is quite low (about 4,000 cells per ml on average; maximum cell number: 13,000 in 1998 and 10,000 in 1999). The mean epilimnic chlorophyll content (Chl a was 10 mg/l in 1998 and 11 g/l in 1999, and the maximum Chl a content was 16 and 22 g/l in 1998 and 1999, respectively.Phytoplankton production can be limited by nutrients, mainly nitrogen, but convective currents can also cause a significant loss of biomass. The lake's euphotic zone is smaller than its epilimnic zone, indicating that light radiation is limiting in the deeper water body, this is caused by a weak thermocline due to destratification by nocturnal cooling, the atelomixis.     

“Man and Biosphere” — Studies on Sikkim Himalayas. Part 2: Acute Toxity of Mixed Copper-Zinc Solutions on Common Carp,Cyprinus carpio (LINN.)

In batch experiments for 6 to 48 h, individuals of Cyprinus carpio (2 g individual weight) were exposed to mixed solutions of copper and zinc sulphate: pH = 6.3, temperature 15 °C, 6.8 mg/l O₂, 7.1 mg/l Ca²⁺, 0.7 mg/l Mg²⁺. The LC₅₀ for the individual substances on exposures of 24 and 48 h amount to 9.04 and 7.28 mg/l, resp., of Zn and 0.11 and 0.095 mg/l, resp., of Cu. The sum of the biological activity of the two kinds of metal ions is assessed according to MARKING'S index: at low concentrations of the mixed solutions the two metals show an additive effect, at higher concentrations a synergistic effect occurs.     

Toxicity of Phenol,Pentachlorophenol and Sodium Pentachlorophenolate to a Freshwater Teleost Lebistes reticulatus (Peters)

Lebistes reticulatus (Peters) was exposed to various concentrations of phenol, pentachlorophenol and sodium pentachlorophenolate to determine their toxicity, lethal concentrations and slope functions by a static bioassay procedure. Simultaneous controls were also run. The 94h LC₅₀ values were 47.5mg/l (42.04 … 53.68) for phenol, 0.77 mg/l (0.616 … 0.963) for sodium pentachlorophenate and 0.97 mg/l (0.866 … 1.086) for pentachlorophenol. On this basis, the rank order is sodium pentachlorophenolate-pentachlorophenol-phenol. During the treatment, fish showed an abnormal swimming reflex, excessive mucus secretion, haemorrhage near lips, abdomen and basis of fins and loss of equilibrium followed by death.     

Zytotoxikologische Untersuchung von Schadstoffen und Wasserblüten mit gefrierkonserviertem Bullensperma (Spermatest)

The method is based on the determination of the number of the nonmotile sperms and/or the lethally damaged sperms with the aid of the fluorescent dye primuline. Incubation takes place at a suspension density of 10.000/mm³… 38.000/mm³, at 40°C or 46 °C for 15… 60 min. During the test with sublimate the loss of motility amounts to 55% at 4 mg/l Hg²⁺, the LC₅₀ is 7.3 mg/l Hg²⁺. Phenyl-mercuric acetate causes the total loss of motility with 20 mg/l, the LC₅₀ is above 168 mg/l (equivalent to 100 mg/l Hg²⁺). The LC₅₀ of sodiumpentadecylmonosulphonate and dodecylpyridinchloride are 11.6 and 15.8 mg/l, resp. Filtrates of blooming of cyanophyceae on Baltic, having been digested by repeated freezing and thawing, with 1.5… 6.4 g/l dry matter showed motility losses of 30… 100 % for Nodularia spumigena, whereas no significant effect was produced by a filtrate of Microcystis aeruginosa with 28.2 g/l dry matter as well as water-blooming of Nodularia spumigena from the Small Jasmund Bodden.     

Immunotoxicity of the xenoestrogen 4-nonylphenol to the cockle Cerastoderma glaucum

The in vivo effects of 4-nonylphenol (NP) on functional responses of haemocytes from the cockle Cerastoderma glaucum were investigated after 7 days exposure to sublethal NP concentrations (0, 0+acetone, 0.0125, 0.025, 0.05 and 0.1 mg/l NP). Haemocytes from both controls and exposed cockles were collected, and the effects of NP on total haemocyte count (THC) and volume of circulating cells, intracellular superoxide anion (O(2)(-)) levels, acid phosphatase and lysozyme-like activities in both haemocyte lysate (HL) and cell-free haemolymph (CFH) were evaluated. Exposure of cockles to 0.1mg/l NP significantly increased THC (p<0.05) with respect to controls. Analysis of haemocyte size frequency distribution showed that the haemocyte fraction of about 7-8 microm in diameter and 250 femtolitres in volume increased markedly in cockles exposed to the highest NP concentration tested. Apoptosis resulting in cell volume reduction in NP-exposed animals cannot be excluded. No statistically significant variation in intracellular O(2)(-) levels was observed. Conversely, significant increases (p<0.05) in acid phosphatase activity were observed in CFH from 0.05 and 0.1mg/l NP-exposed animals; no significant differences in enzyme activity were recorded in HL. Lysozyme-like activity also increased significantly in CFH from cockles exposed to 0.05 mg/l NP (p<0.05) and 0.1 mg/l NP (p<0.001). Instead, lysozyme-like activity decreased significantly (p<0.05) in the HL of animals exposed to 0.05 mg/lNP. Our results suggest that NP induces variations in the functional responses of haemocytes of C. glaucum, mainly by reducing cell membrane stability and promoting cell degranulation.     

The Accumulation of Monocrotophos in Some Piscine Organs

Monocrotophos (0,0-dimethyl-0-(1-methyl-2-N-methyl-carbamoyl)-cis-vinylphosphate) as an active component of some insecticides is highly soluble in water at a low hydrolysis rate: half-time at 2 mg/l, pH = 7.0 and 38 °C 23 d. Individuals of the following fish species (individual weight in g) were kept during the static bioassay for 6… 72 h at 40 mg/l monocrotophos in the medium: Cyprinus carpio (70), Carassius carassius (45) and Leucaspius delineatus (1.5), and then the concentration of monocrotophos was gas-chromatographically determined in different tissues. Initially, till the 24th hour a high accumulation in the gills took place (in mg/kg fresh weight: Cyprinus 0.49, Carassius 3.88), then the concentration remained uniform or slightly falling here, whereas in the other organs (digestive organs, muscle, gonads) a further accumulation up to 1 mg/kg occurred. Leucaspius was investigated only as whole individuals, and after 72 h their fresh weight had increased to 6.96 mg/kg. The rates of accumulation decrease in the following order: Cyprinus: digestive organs, gills, muscle; Carassius: gills, digestive organs, gonads, muscle.     

“Man and the Biosphere”-Studies on Sikkim Himalayas. Part 1: Acute Toxicity of Copper and Zinc to Common Carp Cyprinus carpio (Linn.) in Soft Water

In the daily exchange of the medium with only 7.1 mg/l Ca²⁺ and 0.7 mg/l Mg²⁺ during the batch experiment the LC_50,96h for Cyprinus carpio is 0.063 mg/l Cu²⁺ and 3.12 mg/l Zn²⁺ at water temperatures of 11 to 14°C. Significant differences in mortality as referred to the LC_50,12h occur for copper only after 72 h of exposure and for zinc after 48 h of exposure. The values of LC_50,96h are clearly lower for both metals in the investigations described than most data given in literature for different fish species.     

Biocides in Relation to Water Pollution Part I: Bioassay Studies on the Effects of a few Biocides on Fresh Water Fish,Channa gachua

• - In the present paper the toxicity of four biocides, endrin, thiodon, malathion and phosvel, has been measured by performing static bioassay experiments with Channa gachua as test fish to establish the median Tolerance Limits (TL_m and TL₅₀).
• - The results of bioassay experiments indicate that the fish is harmed even at a very low concentration of these biocides. The values of TL_m are 0.0076, 0.0073, 0.00695 and 0.0068 for endrin; 0.01120, 0.01088, 0.01069 and 0.01064 for thiodon; 9.20, 8.10, 7.90 and 7.60 for malathion and 32.90, 32.10, 31.65 and 31.15 mg/l for phosvel for the time interval of 24, 48, 76 and 96 hours, respectively.
• - As indicated by TL_m's, endrin is most toxic and phosvel least. Thiodon and malathion came in a descending order between endrin and phosvel.
• - During bioassay studies, the effect of some variables, like temerature, pH and fish-size, on TL_m's has also been observed. They gave the following results.
• a) An increase in temperature increased the toxicity of all the biocides.
• b) An increase in pH decreased the TL_m as such, making the biocides more toxic to Channa gachua.
• c) For larger fishes, the biocide solutions were less toxic, but the same solutions were more toxic to fishes of a smaller size.
• - The safe harmless concentrations of endrin, thiodon, malathion and phosvel are - 0.00289 mg/l, 0.00337 mg/l, 1.88365 mg/l and 9.16736 mg/l, respectively.

     

Effect of Zinc,Copper and Mercury on Channa marulius (HAMILTON)

Static bioassay acute toxicity tests of Zinc, Copper and Mercury were conducted to determine the median lethal concentrations (LC₅₀s) of a freshwater teleost Channa marulius (HAM .) The 96 h LC₅₀ and 95% confidence limits for Zn²⁺ were 25.61 (24.13 … 27.12) mg/l; 0.90 (0.80 … 1.038) mg/l; for Cu²⁺ and 0.314 (0.257 … 0.371) mg/l for Hg²⁺. However, these values decreased at 240 h of exposure and were: 21.09 (18.29 … 24.60) mg Zn²⁺/l; 0.66 (0.568 … 0.841) mg Cu²⁺/l; and 0.131 (0.103 … 0.158) mg Hg²⁺/l. The relative potency ratio of Zn/Hg, Zn/Cu and Cu/Hg suggests that fish were most sensitive to Hg, followed by Cu and Zn ions. The acute toxicities of mixtures of Zn²⁺?Cu²⁺; Zn²⁺?Hg²⁺, Cu²⁺?Hg²⁺ and Zn²⁺?Cu²⁺?Hg²⁺ up to 48 h of exposure were also investigated. The additive index and ranges for Zn²⁺?Cu²⁺ were ?0.241 (-0.577 … 0.054); 0.056 (-0.269 … 0.475) for Zn-Hg; 0.285 (-0.043 … 0.718) for Cu-Hg; and -0.542 (-1.215 … 0.005) for Zn-Cu-Hg. All the mixtures tested showed a greater than additive toxicity because index ranges overlapped zero.     

Acute and chronic toxic effects of the carbamate pesticide Sevin on some haematological,biochemical and enzymatic parameters in the fresh water teleost fish Channa punctatus

The effects of exposure to LC₅₀ (10.5 mg/l) for 96 h and to a sublethal concentration (1.05 mg/l) of Sevin for 120 days have been observed on some haematological, biochemical and enzymatic parameters of Channa punctatus. The fish were hyperglycemic and hyperlactemic and the glycogen content of the liver and muscles decreased, but the lactic acid content of two tissues increased. The activity of hexokinase increased in the kidney, intestine, liver and muscles but decreased in the gills and brain. The activity of lactate dehydrogenase decreased in the kidney and intestine, and increased in the liver, muscles, brain and gills. The pyruvate dehydrogenase activity decreased in all the six tissues. Both the exposures produced a decrease in the succinate dehydrogenase activity of the liver. The enzyme activity decreased in the brain, gills and muscles after chronic exposure, whereas the kidney and intestine showed an elevation in enzyme activity.     

Zur Rolle der Stickstoffixation im Stoffhaushalt eines geschichteten Sees

In the dimict lake Arend (5.1 km², 146 hm³, 49.5 m z_max), nitrogen is production-limiting with concentrations below the detection limit during the production period. Phytoplankton achieves biomasses of up to 18 mg/l fresh matter, essential contributions being made by Aphanizomenon with 2 mg/l and Anabaena with up to 10 mg/l. Nitrogen fixation was measured by the ethine reduction technique (acetylene reduction) during periods of the occurrence of heterocystforming Cyanophyceae and achieved peak values up to 6.59 μg N² · h^?1 · l^?1 or 14.87 m^?2 · h^?1 g N₂ · m^?2 · h^?1. The rates of fixation show a safe correlation with the biomass of heterocyst-containing Cyanophyceae (r = 0.88), their development beginning at values below the N : P-ratio of 2.66.     

Hydrogeochemistry of seepage water collected within the Youngcheon diversion tunnel,Korea: source and evolution of SO4‐rich groundwater in sedimentary terrain

In the Youngcheon Diversion Tunnel area, South Korea, 46 samples of tunnel seepage water (TSW) and borehole groundwater were collected from areas with sedimentary rocks (mainly sandstone and shale) and were examined for hydrogeochemical characteristics. The measured SO₄ concentrations range widely from 7·7 to 942·0 mg/l, and exceed the Korean Drinking Water Standard (200 mg/l) in about half the samples. The TDS (total dissolved solid) content generally is high (171–1461 mg/l) from more shale‐rich formations and also reflects varying degrees of water–rock interaction. The water is classified into three groups: Ca? SO₄ type (61% of the samples collected), Ca? SO₄? HCO₃ type (15%) and Ca? HCO₃ type (24%). The Ca? HCO₃ type water (mean concentrations=369 mg/l Ca, 148 mg/l HCO₃ and 23 mg/l SO₄) reflected the simple reaction between CO₂‐recharged water and calcite, whereas the more SO₄‐rich nature of Ca? SO₄ type water (mean concentrations=153 mg/l Ca, 66 mg/l HCO₃ and 416 mg/l SO₄) reflected the oxidation of pyrite in sedimentary rocks and fracture zones. Pyrite oxidation resulted in precipitation of amorphous iron hydroxide locally within the tunnel as well as in high concentrations of Ca (mean 153 mg/l) and Na (mean 49 mg/l) for TSW, and is associated with calcite dissolution resulting in pH buffering. The pyrite oxidation required for the formation of Ca? SO₄ type water was enhanced by the diffusion of oxygenated air through the fractures related to the tunnel's construction. The subsequent outgassing of CO₂ into the tunnel resulted in precipitation of iron‐bearing carbonate. Copyright © 2001 John Wiley & Sons, Ltd.