Effects of dehydroabietic acid and abietic acid on survival, reproduction, and growth of the crustacean Daphnia magna

Resin acids, a class of wood extractives, are potential toxic constituents in many pulp and paper mill effluents. In the present investigation, the effects of two predominant resin acids, dehydroabietic acid (DHA) and abietic acid (ABA), on survival, reproduction, and growth of the freshwater crustacean Daphnia magna were assessed over its life cycle. Based on the experimentally determined acute toxicity data (48-h EC(50)'s) for DHA (7.48 mg/L) and ABA (7.98 mg/L), D. magna was treated chronically with each resin acid at nominal concentrations of 0, 0.25, 0.5, 1.0, 2.0, 4.0, and 8.0mg/L for 21 days. Both DHA and ABA at concentrations as high as 4.0mg/L did not affect physiological and reproductive parameters such as time to maturation, number of molting, number of broods, and number of offspring produced from surviving daphnids, while significant mortality was observed only at 8.0mg/L in both cases. However, a small but statistically significant decrease in Daphnia growth (body length) at the end of exposure was detected at concentrations as low as 0.5mg/L for DHA and 1.0mg/L for ABA, respectively. These results indicated that both DHA and ABA had the potential to exhibit weak growth inhibition without apparent negative effects on reproduction to D. magna at nonlethal concentration levels. This slight effect is not expected to be ecologically significant because the concentrations of DHA and ABA in biologically treated pulp and paper mill effluents are well below the effective levels observed in the present study.     

Use of Japanese Medaka (Oryzias latipes) and Tilapia (Oreochromis mossambicus) in Toxicity Tests on Different Industrial Effluents in Taiwan

In Taiwan, aquatic toxicity tests for industrial effluents are not required for discharge permits. However, relying on traditional chemical and physical characteristics of an effluent to monitor and regulate such discharges to manage water quality of a receiving water is insufficient. In this study, we used two fish species, Japanese medaka (Oryzias latipes) and tilapia (Oreochromis mossambicus), and three toxic endpoints, including acute and subacute toxicity, to determine toxicity of seven different types of industrial effluents. Prior to the study, two reference toxicants were tested on two fish species. The LC50s of CdCl₂ for tilapia and medaka juveniles were 29.6 ± 15.3 mg/L and 2.2 ± 1.2 mg/L, respectively. The sensitivity of medaka embryo mortality and hatching inhibition to CdCl₂ were about the same, with the LC50 and EC50 of 0.3 ± 0.1 mg/L and 0.1 ± 0.1 mg/L, respectively. The LC50s for tilapia and medaka juveniles to sodium dodecyl sulfate (SDS) were 19.7 ± 10.6 mg/L and 12.5 ± 5.9 mg/L. The medaka embryo was less sensitive to SDS than to CdCl₂. The embryo's LC50 for SDS was 5.8 ± 2.8 mg/L and the hatching inhibition EC50 was 1.3 ± 1.1 mg/L. Results of toxicity tests on different effluents showed that the electroplating effluent was the most toxic, followed by acrylonitrile manufacturing and pulp/paper mill discharges. The LC50s of the electroplating effluent to different assays were in the range of several percents of the whole effluent. The pulp/paper effluent was toxic only to the medaka embryo. The rest of the industrial effluents tested showed either moderate or no toxicity to the animals. Received: 12 June 2000/Accepted: 6 September 2000     

Characterization of putative ligands for a fish gonadal androgen receptor in a pulp mill effluent

Fish exposed to pulp and paper mill effluents often become masculinized. A plausible hypothesis is that this is caused by activation of androgen receptors. The present study aimed to investigate if ligands for the fish gonadal androgen receptor (AR2) are present in pulp mill effluent and to characterize/identify these compounds. Extracts of both primary and biologically treated effluents from a Swedish kraft pulp mill were fractionated chemically. Fractions were tested in competitive binding assays for AR2 from ovaries of the Atlantic croaker (Micropogonias undulatus). Primary effluent contained 96 ng dihydrotestosterone equivalents/ L, whereas biologically treated effluent was 16 times less potent. Further fractionations and assays of binding activities were performed on the primary effluent. Eight final fractions displaced androgen in the binding assay, and gas chromatography/mass spectrometry (GC/MS) analyses revealed that these contained 37 detectable compounds that were not present in inactive fractions. The majority were moderately polar compounds between 200 and 400 g/mol with hydroxyl/carbonyl groups. Two compounds were ruled out because of their lack of binding to AR2. The mass spectra of a third compound matched that of 4-hydroxy-3 (2-(4-hydroxy-3-methoxophenyl)ethyl)-5-metoxyacetophenon, but the remaining candidates could not be fully identified. A search for 21 known steroidal AR2 ligands showed that progesterone, a relatively strong AR2 ligand, was present in the primary effluent (1.6 microg/L) but was removed during the biological treatment step. The detection of multiple fractions with significant binding activity indicates that a variety of compounds in effluents have the potential to masculinize fish near pulp mills via an androgen receptor-mediated mechanism.     

Expression of the vitellogenin gene in the liver of juvenile whitefish (Coregonus lavaretus L. s.l.) exposed to effluents from pulp and paper mills.

Juvenile whitefish (Coregonus lavaretus L. s.l.) were exposed by caging in the field to diluted effluents from three operating pulp, paper, and paperboard mills in Southern Lake Saimaa, Finland. The expression of the vitellogenin gene, used as a biomarker of estrogenic contamination of effluents, was measured using a Northern blotting method. Increased mRNA levels, the most specific and reliable evidence for estrogen receptor-mediated actions in vivo, were found in fish caged in the vicinity of one of three mills studied. This mill was found to discharge wood-derived compounds, such as sterols and resin acids, into Lake Saimaa in amounts considerably exceeding those from the other two mills. The increased vitellogenin gene expression suggests that the effluent is a source of estrogenic contaminants.     

Biomarker Responses in Whitefish (Coregonus lavaretus L. s.l.) Experimentally Exposed in a Large Lake Receiving Effluents from Pulp and Paper Industry

Physiological and biochemical biomarker responses were studied in juvenile whitefish (Coregonus lavaretus L. s.l.) exposed experimentally to effluent from the forest industry. The large study area (609 km²), Southern Lake Saimaa, in Southeast Finland, receives 330,000 m³ d⁻¹ of biologically and 55,000 m³ d⁻¹ of chemically treated effluents, discharged from two integrated elementary chlorine free (ECF) bleached kraft pulp and paper mills, from one ECF pulp mill, and from one mill producing unbleached pulp and cardboard. The assessment of exposure to effluent discharged from the mills was based on lake water chlorophenolics (CPs) and resin acids (RAs) measured in samples collected from the 22 experimental sites along the area. Despite the low levels of effluent constituents in the lake, they were still accumulated in detectable levels in fish bile, indicating an exposure to the bioactive compounds of effluents. In comparison to the reference area, a two- to four-fold increase in ethoxyresorufin O-deethylase (EROD) activity was observed in whitefish exposed in the vicinity (1–6 km) of all the mills. However, cytochrome P450 1A1 (CYP1A1) gene expression was increased in only one of the receiving areas, indicating higher sensitivity of the EROD activity in the present study. There were no statistically significant correlations between EROD activity and the ambient water concentrations of the CPs, the RAs, or effluent dilution expressed by water sodium concentration. Neither bile chlorophenolics nor bile resin acids showed a significant correlation with EROD. No significant changes in circulating reproductive steroids, 17β-estradiol and testosterone, in juvenile whitefish were observed. The vitellogenin gene was expressed in the vicinity of the pulp mill discharging the most wood-derived compounds, i.e. resin acids and wood-sterols, including β-sitosterol. No differences were observed in plasma immunoglobulin M, glucose, or lactate concentrations between the effluent sources. Received: 25 March 1997/Accepted: 28 July 1997     

Selected resin acids in effluent and receiving waters derived from a bleached and unbleached kraft pulp and paper mill

Water samples were collected on three dates at 24 sites influenced by effluent from Georgia-Pacific's Palatka Pulp and Paper Mill Operation, a bleached and unbleached kraft mill near Palatka, Florida, USA. The sampling sites were located within the mill retention ponds, Rice Creek, and the St. John's River. Samples were analyzed by gas chromatography-mass spectrometry for abietic, dehydroabietic, and isopimaric acids, all of which are potentially toxic by-products of pulp production. Isopimaric acid concentrations greater than 12 mg/L were measured at the mill's effluent outfall but were less than 20 microg/L at the end of Rice Creek. This result indicates that the waters of Rice Creek provide dilution or conditions conducive for degradation or sorption of these compounds. Large differences in resin acid concentrations were observed between sampling events. In two sampling events, the maximum observed concentrations were less than 2 mg/L for each analyte. In a third sampling event, all of the compounds were detected at concentrations greater than 10 mg/L. Data from the three sample dates showed that resin acid concentrations were below 20 microg/L before the confluence of Rice Creek and the St. John's River in all cases.     

Use of paired fathead minnow (Pimephales promelas) reproductive test. Part 2: source identification of biological effects at a bleached kraft pulp mill

Reproductive effects of pulp mill effluents on fish continue to be reported in Canadian waters. Spawning delays, reduced gonad size, and altered levels of sex steroid hormones have been found in both sexes of various fish species exposed to effluents. We initiated a project to identify the source/cause of such effects. In part 1 of this two-part series, we exposed mature adult fathead minnow (FHM; Pimephales promelas) for 21 d to final treated effluent (1% and 100% v/v) from a bleached kraft pulp mill in Terrace Bay (ON, Canada). Results suggested pulp mill effluent from this mill affected reproductive indicators in FHM and effects were dependent on effluent concentration, duration of exposure, and method of data analysis. The main objective of this paper was to use the FHM assay to identify waste stream sources within the mill that affect reproductive indicators. Various process streams were selected, characterized with respect to effluent chemistry and acute toxicity, and a subset was tested on-site with the 21-d FHM bioassay. Results showed that both the combined mill effluent (before secondary treatment) and the combined alkaline stream (CALK) caused both decreased spawning events (approximately 55% for both streams) and decreased egg production (28 and 74%, respectively), and the CALK stream resulted in significant male ovipositor development. By comparing response patterns we were able to identify the CALK stream as a source of compounds at this mill affecting reproductive indicators in FHM and highlight this stream for further toxicity identification evaluation.     

Ecotoxicological characterization of industrial wastewater--sulfite pulp mill with bleaching

Different process wastewaters from a sulfite pulp mill with bleaching were characterized by chemical analysis and toxicity tests. The amount of adsorbable organically bound halogen (AOX) from the bleachery was 3.6 kg per ton pulp. The extractable organically bound chlorine was 15% of AOX. Some identified organochlorine compounds in the effluent could be traced in the receiving water. Effluents from the chlorination and alkaline extraction stages and the condensate were the main contributors to the effluent toxicity. The effluents were particularly toxic to the alga Skeletonema costatum. The EC50 value for growth of the alga was 24-29 ml/liter of the total effluent. The toxicity cannot be ascribed to single chemical components in the effluents. Degradation of toxic components occurs after dilution of the effluents in the receiving water. The toxicity may be reduced to 30-50% of the initial toxicity within 1 week. Predictions of toxic effects in the receiving water, based on results of toxicity tests and estimated dilution, indicate that large areas are affected by the discharges. Som observations of the distribution of organisms in the receiving water indicate that predictions from the toxicity tests may be valid.     

Ecotoxicity and genotoxicity assessment of cytostatic pharmaceuticals

The fate and effects of cytostatic (anticancer or antineoplastic) pharmaceuticals in the environment are largely unknown, but they can contaminate wastewater treatment effluents and consequently aquatic ecosystems. In this paper, we have focused on five cytostatic compounds used in high amounts (cyclophosphamide, cisplatin, 5-fluorouracil, doxorubicin, and etoposide), and we have investigated their ecotoxicity in bacterial Pseudomonas putida growth-inhibition test, algal Pseudokirchneriella subcapitata growth-inhibition test, and Dapnia magna acute immobilization test. Genotoxicity also was assessed with Escherichia coli SOS-chromotest (with and without metabolic activation) and the GreenScreen Assay using yeast S. cerevisiae. All tested compounds showed significant effects in most of the assays with lowest-observed-effect concentrations and concentrations causing 50% effects (EC50s) values ranging within microg/L to mg/L. The most toxic compound was 5-fluorouracil in the assays with P. putida (EC50 = 0.027 mg/L) and P. subcapitata (EC50 = 0.11 mg/L), although cisplatin and doxorubicin were the most toxic to D. magna (EC50 = 0.64 and 2.0 mg/L, respectively). These two chemicals were also the most genotoxic in the SOS-chromotest (minimum genotoxic concentrations [MGC] = 0.07-0.2 mg/L), and 5-fluorouracil was the most genotoxic in the eukaryotic yeast assay (MGC = 0.02 mg/L). Our investigation seems to indicate generally lower risks of acute effects at concentrations expected in the environment. However, some effective concentrations were relatively low and chronic toxicity of cytostatics (and/or their transformation products), as well as specific sources of human pharmaceuticals such as hospital effluents, require research attention.     

Bioavailability models for predicting acute and chronic toxicity of zinc to algae, daphnids, and fish in natural surface waters

Bioavailability models predicting acute and/or chronic zinc toxicity to a green alga (Pseudokirchneriella subcapitata), a crustacean (Daphnia magna), and a fish (Oncorhynchus mykiss) were evaluated in a series of experiments with spiked natural surface waters. The eight selected freshwater samples had varying levels of bioavailability modifying parameters: pH (5.7-8.4), dissolved organic carbon (DOC, 2.48-22.9 mg/L), Ca (1.5-80 mg/L), Mg (0.79-18 mg/L), and Na (3.8-120 mg/L). In those waters, chronic zinc toxicity (expressed as 10% effective concentrations [EC10]) varied up to 20-fold for the alga (72-h EC10 from 27.3 to 563 microg Zn/L), and approximately sixfold for the crustacean (21-d EC10 from 59.2 to 387 microg Zn/L), and fivefold for the fish (30-d LC10, lethal concentration for 10% of the organisms, from 185 to 902 microg Zn/L). For P. subcapitata a refined bioavailability model was developed by linking an empirical equation, which predicts toxicity expressed as free Zn2+ activity as a function of pH, to the geochemical speciation model WHAM/Model V. This model and previously developed acute and/or chronic biotic ligand models for D. magna and 0. mykiss generally predicted most effect concentrations by an error of less than a factor of two. In waters with pH > 8, however, chronic toxicity to D. magna was underestimated by a factor 3 to 4. Based on the results of this validation exercise and earlier research, we determined applicability ranges for pH (6-8) and Ca (5-160 mg/L) in which all three developed models are valid. Within these ranges, all three models may be considered useful tools for taking into account bioavailability in regulatory assessments of zinc.     

Kraft mill effluent survey: progress toward best management practices for reducing effects on fish reproduction

Pulp and paper mill effluents have been linked to effects on fish reproduction for more than 25 years. To date, the causes of these effects and remedial strategies have eluded investigators. Recent work has shown that the degree of reproductive effect caused by a mill effluent is related to the overall organic content. If verified, this could lead to breakthroughs for best management practices (BMPs). For this study, the effluents from seven kraft mills were assessed for their ability to reduce egg production in fathead minnow (Pimephales promelas) in the laboratory. At the same time, the effluents were analyzed for three parameters thought to be good indicators of organic losses: biochemical oxygen demand (BOD), methyl-substituted 2-cyclopentenones (cellulose degradation products), and a gas chromatographic (GC) profile index, which integrates the total area of the chromatographic peaks of solvent-extracted effluents using low-resolution mass spectrometry. The results showed that the degree to which the effluents reduced egg production increased with increased organic losses as characterized by BOD and the GC profile index. Therefore, these parameters could be used to guide BMPs at kraft mills according to specific targets: BOD?相似文献   

Chronic Effects of <Emphasis Type="Italic">Pinus radiata</Emphasis> and <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> Kraft Mill Effluents and Phytosterols on <Emphasis Type="Italic">Daphnia magna</Emphasis>

Two kraft pulp mill effluents were compared in terms of their chronic toxicity to Daphnia magna. One resulted from pulping Pinus radiata and the other came from a parallel processing of Pinus radiata and Eucalyptus globulus (mixed kraft pulp mill effluent). The concentration of phytosterols found in the mixed kraft pulp mill effluent was higher than in the effluent from Pinus radiata, with values of 0.1082 and 0.02 μg/L, respectively. The phytosterols per se are responsible for 12.9% and 8.1% of the deviation from the natural shape, while the kraft pulp mill effluents account for 25.6%–27.8% of shape deviation. The role of β-sitosterol and stigmasterol is discussed in relation to endocrine disruption.     

Toxicity of laundry detergent components to a freshwater cladoceran and their contribution to detergent toxicity

The toxicity of 39 laundry detergent components including surfactants, enzymes, builders, fabric brighteners, fillers, and coloring agents to the cladoceran Ceriodaphnia cf. dubia was determined. The difference between the most and the least toxic components was approximately 17,000-fold and 1,000,000-fold for the mg/L and mmol/L EC50 data, respectively. Two of the components had high toxicity (EC50 values < 1 mg/L), 11 moderate toxicity (EC50 values between 1 and 10 mg/L), and the remaining 26 components had low toxicity (EC50 values > 10 mg/L). Analysis revealed that mixtures of the components interacted antagonistically, additively, and synergistically. On a molarity basis the most toxic group of compounds was the surfactants followed by the brighteners. The most toxic individual components included sodium carboxymethyl cellulose, sodium silicate solution, four brighteners, sodium perborate tetrahydrate, and the surfactants. Many of the most toxic components, however, contributed very little to the toxicity of the detergents due to being present in the detergents at low concentrations. The main contributors to the toxicity of detergents were the sodium silicate solution and the surfactants-with the remainder of the components contributing very little to detergent toxicity. The potential for acute aquatic toxic effects due to the release of secondary or tertiary sewage effluents containing the breakdown products of laundry detergents may frequently be low. However, untreated or primary treated effluents containing detergents may pose a problem. Chronic and/or other sublethal effects that were not examined in this study may also pose a problem.     

Determination of degradation products of alkylphenol polyethoxylates in municipal wastewaters and rivers in Tokyo, Japan

An analytical method for the simultaneous determination of nonylphenoxy acetic acid (NP1EC), nonylphenol monoethoxy acetic acid (NP2EC), nonylphenol (NP), octylphenol (OP), and nonylphenol monoethoxylate (NP1EO) was developed. The method was applied to environmental samples to demonstrate the distribution and behavior of nonylphenol polyethoxylates and their degradation intermediates in aquatic environments in Tokyo, Japan. In sewage treatment plants, more than 85% of NP, OP, and NP1EO were removed, whereas NP1EC and NP2EC were generated during the treatment. Concentrations of NP1EC and NP2EC in secondary effluents (1.9-2.9 microg/L) were higher than those of NP and NPIEO (0.12-0.63 microg/L). In river waters. NPIEC and NP2EC concentrations (0.11-2.8 microg/L) were higher than NP and NP1EO concentrations (<0.015-3.4 microg/L). with some exceptions. In surface sediments, neither NPIEC nor NP2EC was detectable (<0.01 microg/g dry wt) whereas NP and NPIEO were detected significantly (0.03-1.82 microg/g dry wt and 0.04-0.46 microg/g dry wt, respectively).     

Identification and treatment of a waste stream at a bleached-kraft pulp mill that depresses a sex steroid in the mummichog (Fundulus heteroclitus)

Changes to indicators of reproductive performance have been documented in fish exposed to some bleached kraft pulp mill effluents (BKPMEs). However, responses are not consistent across mill types or processes. It is not clear where the sources of the effects are within mills, what the causative compounds are, or what process changes are effective to remove these sources. Our previous studies suggested that condensates were a source of compounds that reduced plasma testosterone in the mummichog (Fundulus heteroclitus). Results also suggested that reverse osmosis (RO) treatment of condensates removed this source. The objective of this study was to use a toxicity identification evaluation approach and expose mummichog to various waste streams in the laboratory to identify the effluent source that depressed plasma testosterone in the mummichog and the effects of RO treatment. In 7- and 21-d exposures, mummichog were exposed to dilutions of RO feed condensate, RO permeate, combined mill effluent (CME), and final effluent. Results confirmed that condensates depressed plasma testosterone in mummichog. Chemical characterization of the condensate indicated that plant phytosterols were likely not the responsible compounds. Results also confirmed that RO treatment removed the potential of the condensates to depress plasma testosterone in mummichog at environmentally relevant concentrations of final mill effluent (1%).     

Ecotoxicological risk of pharmaceuticals from wastewater treatment plants in Korea: occurrence and toxicity to Daphnia magna

The overall ecotoxicological effect of pharmaceutically active compounds (PhACs) detected in the effluents of Korean wastewater treatment plants (WWTPs) to Daphnia magna was investigated using biological and chemical analyses. The bioassay results showed median lethal concentrations and no-observed-effect concentrations ranging from a few to tens of ppm levels for nine PhACs in 48-h acute and 21-d chronic tests. The mixture effects of pharmaceuticals also were examined by other acute and chronic tests, which showed no significant toxicity despite a slightly increased combined effect of approximately twofold. The residual concentrations of nine PhACs were analyzed at concentrations ranging from 10 ng/L to 89 microg/L in the influents and from 10 ng/L to 11 microg/L in the effluents from four metropolitan cities in South Korea between January and November of 2004. Through repeated investigations of the influents and the effluents from different WWTPs, relatively higher removal efficiencies (23.9-91.3%) compared with those of previous surveys performed in other countries were observed for most pharmaceuticals, with the exception of acetaminophen (8.7%). The present study showed no significant risk effects of the effluents from WWTPs containing pharmaceuticals (i.e., hazard quotient < 1), even at the 95th percentile contamination range, although a risk assessment factor of 1,000 was applied. Therefore, it can be concluded that the potential risk of pharmaceuticals should be monitored carefully with more bioassay data, because many uncertainties still exist in the determination and toxicity of metabolites in water environments. No significant risk was observed, however, from the selected PhACs in the effluents from WWTPs discharged into surface waters.     

Assessment of hormonal activities and genotoxicity of industrial effluents using in vitro bioassays combined with chemical analysis

Wastewaters from various industries are a main source of the contaminants in aquatic environments. The authors evaluated the hormonal activities (estrogenic/anti-estrogenic activities, androgenic/anti-androgenic activities) and genotoxicity of various effluents from textile and dyeing plants, electronic and electroplate factories, pulp and paper mills, fine chemical factories, and municipal wastewater treatment plants in the Pearl River Delta region by using in vitro bioassays (yeast estrogen screen [YES]; yeast androgen screen [YAS]; and genotoxicity assay [umu/SOS]) combined with chemical analysis. The results demonstrated the presence of estrogenic, anti-estrogenic, and anti-androgenic activity in most industrial effluents, whereas no androgenic activities were detected in all of the effluents. The measured estrogenic activities expressed as estradiol equivalent concentrations (EEQs) ranged from below detection (3 of 26 samples) to 40.7 ng/L, with a mean of 7.33 ng/L in all effluents. A good linear relationship was found between the EEQs measured by YES bioassay and the EEQs calculated from chemical concentrations. These detected estrogenic compounds, such as 4-nonylphenol and estrone, were responsible for the estrogenic activities in the effluents. The genotoxic effects expressed as benzo[a]pyrene equivalent concentrations (BaP EQs) varied between below detection and 88.2 μg/L, with a mean of 8.76 μg/L in all effluents. The target polycyclic aromatic hydrocarbons were minor contributors to the genotoxicity in the effluents, and some nontarget compounds in the effluents were responsible for the measured genotoxicity. In terms of estrogenic activities and genotoxicity, discharge of these effluents could pose high risks to aquatic organisms in the receiving environments.     

Male-biased sex ratios of fish embryos near a pulp mill: temporary recovery after a short-term shutdown

In a previous study we showed that broods from the viviparous eelpout Zoarces viviparus were significantly male biased in 1998 in the vicinity of a large kraft pulp mill on the Swedish Baltic coast. One suggested hypothesis was that masculinizing compounds in the effluent were affecting gonadal differentiation of the embryos, resulting in skewed sex ratios. In this article, we present further evidence for a causal relationship between the exposure to the effluent and the male-biased sex ratios. Analyses of historical samples showed that the eelpout produced male-biased broods close to the mill in 1997 in addition to 1998. During 1999, the mill was shut down for 17 days, coinciding with the period when the gonads of the eelpout embryos differentiate. Subsequently, in the fall of 1999, the sex ratios were no longer male biased; however, the following year (2000), a significant male bias reappeared. Investigations at 13 sites for up to 4 years showed a relatively stable sex ratio around 50/50, with the exceptions by the mill and with few observations of deviating ratios at other sites. Several reports document endocrine disturbances in fish near pulp and paper mills, including the expression of male secondary sex characters in female fish. The repeatedly identified male bias at the investigated mill, the normalization after mill shutdown, and the reappearance the following year indicate that pulp mill effluents also can affect sex ratios of nearby fish.     

Reproductive, physiological, and biochemical responses in juvenile female rainbow trout (Oncorhynchus mykiss) exposed to sediment from pulp and paper mill industrial discharge areas

Four pulp and paper mills discharge their effluents in the same section of the Biobio River in central southern Chile. Pulp mill effluents are a very complex mixture with characteristics that depend on the type of raw material, the process technology, and the effluent treatment. To investigate the effect of pulp mill effluent discharges, immature Oncorhynchus mykiss were exposed to river sediments in the laboratory for 29 d. Three sampling areas were defined in a spatial gradient in the river: Preimpact, impact, and postimpact zones relative to the pulp and paper mill discharge areas. Ethoxyresorufin-O-deethylase activities were significantly higher in fish exposed to impact and postimpact sediments when compared to those exposed to preimpact sediments, and higher levels of vitellogenin were observed in the plasma of female fish exposed to impact and postimpact sediments. Histological analysis of the gonadal tissue showed an induction of gonadal maturation in fish exposed to sediment coming from the impact and postimpact zones (oocytes in a vitellogenic state). No site differences were observed in erythrocytes, although differences were noted in the leukocytes in the exposure areas. Finally, the biomarker approach showed evidence that the sediment associated with pulp mill effluent discharges produces some effects in fish under laboratory conditions.     

Assessing the performance of Hormosira banksii (Turner) Desicaine germination and growth assay using four reference toxicants

This study evaluated the performance of a Hormosira banksii germination inhibition bioassay developed to assess the toxicity of single compounds and complex effluents in the Australasian region. The reproducibility of 48 and 72 h germination and growth toxicity tests was determined using four reference toxicants, namely, ammonia, copper, phenol and zinc. H. banksii spore germination and growth was significantly inhibited following exposure to each of the toxicants. H. banskii spores were most sensitive to ammonia and showed decreasing sensitivity to copper, zinc and phenol. The EC50 values ranged from 0.07 to 0.08 mg L(-1) for ammonia, 0.09 to 0.22 mg L(-1) for copper, 36 to 1068 mg L(-1) for phenol and 19 to 23 mg L(-1) for zinc. Variability of the test method ranged from 14% to 144% and was comparable to that observed for commonly employed bioassays against appropriate reference toxicants.