Improved understanding of key elements governing the toxicity of energy ash eluates.

Ash from incinerated waste consists mainly of a complex mixture of metals and other inorganic elements and should be classified based on its inherent hazardous effects according to EUs Waste Framework Directive. In a previous study, we classified eight eluates from ash materials from Swedish incineration plants, both chemically and ecotoxicologically (using bacteria, algae, crustacean and fish). Based on measured concentrations in the eluates together with literature acute toxicity data on the crustacean Nitocra spinipes we identified six elements (i.e. Zn, Cu, Pb, Al, K and Ca) potentially responsible for the observed ecotoxicity. However, comparing the used test methods with N. spinipes, the acute test was relatively insensitive to the eluates, whereas the (sub)chronic test (i.e. a partial life cycle test, investigating larval development ratio) was very sensitive. The overall aim of this follow-up study was to verify if the pinpointed elements could be responsible for the observed (sub)chronic toxicity of the eluates. Individual effect levels (i.e. NOEC values) for these six elements were therefore generated using the (sub)chronic test. Our results show that for six of the eight eluates, the observed ecotoxicity can be explained by individual elements not classified as ecotoxic (Al, K and Ca) according to chemical legislation. These elements will not be considered using summation models on elements classified as ecotoxic in solid material for the classification of H-14, but will have significant implications using ecotoxicological test methods for this purpose.

An ecotoxicological approach for hazard identification of energy ash.

Within the EU, ash should be classified by its inherent hazardous effects under criterion H-14 (ecotoxic) in the Directive on waste (2008/98/EC). Today, however, there are no harmonized quantitative criterions for such a classification, but it is stated that biological test systems can be used. In this study seven ash materials were leached and characterized, both biologically and chemically. The objectives were to evaluate if (a) clear concentration-response relationships could be achieved for the selected toxicity tests (bacteria, algae, crustacean and fish), (b) some test(s) are generally more sensitive and (c) the toxic responses were consistent with the chemical analyzes. Interestingly, our results indicate that high concentrations of non-hazardous components (Ca, K) influenced the toxicity of almost all ash eluates, whereas hazardous components (e.g. Zn, Pb) only influenced the toxicity of the eluates ranked as most hazardous. If considering both hazardous and non-hazardous substances, the observed toxic responses were relatively consistent with the chemical analyzes. Our results further showed that the (sub)chronic tests were much more sensitive than the acute tests. However, the use of extrapolation factors to compensate for using the less sensitive acute tests will likely lead to either over- or underestimations of toxicity. Our recommendation is therefore that classification of waste according to H-14 should be based on (sub)chronic test data. Finally, given that treatment of the eluates prior to toxicity testing has a major significance on the concentration and speciation of released substances, further studies are needed in order to propose a relevant testing scheme.

Toxic substances and reproductive disorders in Baltic fish and crustaceans.

In the Baltic Sea ecosystem reproductive disorders have occurred in top consumers such as seals and some fish-eating birds, due to biomagnification of toxic substances, e.g. DDT and PCBs. Reproductive disturbances have also affected fish during the last 25 years. However, there is no strong evidence that toxic substances have caused these problems. Rather, the disorders seem to result from a combination of two or more biotic or abiotic factors. The M74 syndrome, which kills fry of salmon and sea trout, is characterized by a deficiency of thiamine (vitamin B1). Several factors may contribute to the thiamine deficiency, including the diet of salmon in the sea and halogenated organic compounds. Cod do not develop M74, and poor cod recruitment is mainly due to poor oxygen conditions in the spawning areas in combination with overfishing. Toxic compounds in pulp-mill effluents retard gonadal development in perch, but the mechanisms and the active substances have not been identified. Recruitment problems in perch in the coastal waters outside some pulp mills may also be explained by a lack of food items for juvenile fish, rather than reproductive failure. There are very limited data on reproductive disorders in crustaceans from the Baltic Sea. Most data come from studies of the benthic amphipod Monoporeia affinis, which has been used in monitoring programs. Several signs of reproductive disorder have been reported in this amphipod, e.g. malformation and death of embryos, and asynchronous maturation of males and females.

Oestrogens have no hormonal effect on the development and reproduction of the harpacticoid copepod Nitocra spinipes.

In recent years, reports have described endocrine-disruptive effects of environmental oestrogens in fish, but little is known about similar effects in crustaceans. The objective of the present study was therefore to examine whether the oestrogens 17-beta-oestradiol, 17-alpha-ethynylestradiol and diethylstilbestrol (DES), could affect mortality, larval development rate, fecundity and sex ratio in the sexually reproducing harpacticoid copepod Nitocra spinipes. Newly released nauplii (<24-h old) were exposed to 1/1,000, 1/100 and 1/10 (nominal concentrations) of each oestrogen's 96 h-LC50 value for < or = 18 days at 22 +/- 1 degrees C. The percentage of gravid females and the number of developed copepodites were both reduced at 0.03 mg l(-1) DES, although the latter response was not significant. None of the other two oestrogens induced any measurable effects. Since the only observed significant response appeared at a DES concentration no more than 10 times below the 96 h-LC50 value, there is no evidence of endocrine-disruptive activity in N. spinipes exposed to oestrogens.

Chemicals and biological products used in south-east Asian shrimp farming, and their potential impact on the environment--a review.

A wide variety of chemicals and biological products are used to treat the water and sediment of ponds in semi-intensive and intensive south-east Asian shrimp farming. These products are also often used in shrimp hatcheries and to disinfect equipment for shrimp pond management. In spite of the size and importance of the shrimp farming industry in several south-east Asian countries, documentation of the quality and quantity of chemicals and biological products used during farming is scarce. This paper is a compilation of the literature available on substances used in shrimp farming, and the possible environmental effects of these products are analysed to the extent allowed by the limited information. The role of shrimp farm managers, the chemical industry, governments, inter-governmental organisations and scientists in the development of a sustainable practice is discussed. It is concluded that shrimp farmers should reduce the use of chemicals and biological products because of the risks to the environment, human health and to production, and also, because many chemicals and biological products used in pond management have not been scientifically shown to have a positive effect on production. Clearly, the use of some chemicals, i.e. certain antibiotics, poses a risk of danger towards human health. Some chemicals used in shrimp farming, such as organotin compounds, copper compounds, and other compounds with a high affinity to sediments leave persistent, toxic residues, and are likely to have a negative impact on the environment. However, to assess the reality of these risks, substantial new information about the quantity of chemicals used in marine south-east Asian shrimp farming is needed.

Toxicity to Microtox, micro-algae and duckweed of effluents from the Bai Bang paper company (BAPACO), a Vietnamese bleached kraft pulp and paper mill.

Toxicity of the combined effluent as well as effluents from different production units of BAPACO, a Vietnamese bleached kraft integrated pulp and paper mill, was studied using three ecotoxicological tests: Microtox, the micro-alga, Selenastrum capricornutum, and the duckweed, Lemna aequinoctialis. Physico-chemical characteristics of the effluents were also analyzed. Due to unstable operating conditions of the mill, a number of samples taken at different monitoring periods from each effluent were tested, in order to get statistically reliable ranges and averages of toxicity characteristics. For the mill combined effluent, it was found that the micro-algae were the most sensitive followed by Microtox bacteria, while duckweed was not sensitive. Microtox tests showed that the bleaching filtrate from the chlorination stage (C-stage) was the most toxic among all effluents in the mill. The combined effluent from the bleaching plant contributed the largest TEF (Toxicity Emission Factor) to the toxicity balance of the whole BAPACO combined effluent. A less-than-additive mode of toxicity was obtained. In spite of high pollution loads, e.g. BOD, COD, SS, to the Red River, the toxicity of the effluent was considered to be low. However, during periods of low river water sublethal toxic effects on the biota may be expected.

Sublethal effects of tetrachloro-1,2-benzoquinone--a component in bleachery effluents from pulp mills--on vertebral quality and physiological parameters in fourhorn sculpin.

The effects of tetrachloro-1,2-benzoquinone (TCQ), a component in bleached kraft mill effluents (BKME), on vertebral and physiological parameters were investigated in juvenile fourhorn sculpin, Myoxocephalus quadricornis L. After about 4.5 months of exposure to 0.1 and 0.5 mg TCQ/liter in 7% salinity brackish water, the fish demonstrated vertebral deformities, aberrant mechanical properties of the vertebrae, effects on white and red blood cell counts, enhanced activity of delta-aminolevulinic acid dehydratase in erythrocytes, and increased levels of ascorbic acid in the liver. The effects are discussed in relation to those previously observed in fish exposed to complex BKME in the field and the laboratory.

Long-term effects of bleached kraft mill effluents on red and white blood cell status, ion balance, and vertebral structure in fish.

In a laboratory investigation fourhorn sculpin (Myoxocephalus quadricornis) were exposed for 5-9 months to wastewater from pine and birch lines from a bleached kraft pulp plant. This long-term exposure to bleached kraft mill effluent (BKME) affected the hematology, the ion balance, and the vertebral structure. Decreased values for hematocrit and hemoglobin and a reduced red blood cell count, as well as increased levels of methemoglobin, indicated disturbances in the red blood cell status. The white blood cell picture was not significantly affected by the BKME exposure. Decreased levels of potassium and chloride ions in the blood plasma in some of the exposed fish suggest an impaired ability to maintain ion homeostasis. Elevated frequencies of vertebral deformations in fish exposed to BKME confirm previous observations of vertebral damage in feral fourhorn sculpin caught in the receiving body of water of the same bleached kraft pulp industry. Many of the parameters analyzed in this investigation may be used as health indicators in future laboratory and field studies on fish exposed to BKME.

A flowthrough fecundity test with Nitocra spinipes (Harpacticoidea Crustacea) for aquatic toxicity.

A sublethal flowthrough fecundity test with the euryhaline harpacticoid copepod Nitocra spinipes has been developed as a complement to the acute toxicity test (for 48 or 96 hr LC50) with the same species (B-E. Bengtsson, 1981, Mar. Pollut. Biol. 9,238-241). Bacterial suspension as feed and test water are continuously fed by a peristaltic pump to the system. Newly fertilized females with ovigerous bands are harvested from laboratory cultures and put into the test vessels at the start of the experiment. They are then exposed to a series of concentrations of chemicals or industrial effluents for 13 days. The amount of live offspring (metanauplia and copepodids) are recorded and an EC50 for fecundity is calculated. The report gives a detailed technical description of the test system and presents the results from 11 tests with pure chemicals (Zn, Cd, As, and pentachlorophenate) and six industrial effluents (pulp industry, textile industry, and refinery) in salinities ranging from 3 to 25%.

Long-term effects of bleached kraft mill effluents on carbohydrate metabolism and hepatic xenobiotic biotransformation enzymes in fish.

In a laboratory investigation, fourhorn sculpin (Myoxocephalus quadricornis) were exposed for 5-9 months to waste water from pine and birch lines from a bleached kraft pulp plant. The bleached kraft mill effluents (BKME) affected both the carbohydrate metabolism and the xenobiotic metabolism. Thus, an elevated muscle glycogen content in fish exposed to effluent from the pine pulp line suggests a metabolic imbalance. A liver enlargement and a strong elevation of the hepatic cytochrome P-450-dependent ethoxyresorufin-O-deethylase activity after exposure to the bleached pine pulp effluents indicate the presence of cytochrome P-450-inducing agents in the BKME. The results also demonstrate that many physiological test parameters may be used as good indicators of sublethal disturbances in fish to BKME exposure.

The use of chemical and biological parameters to characterize complex industrial effluents.

Hitherto, the concern for possible environmental effects due to aqueous industrial discharges has been directed toward clearly observable effects such as eutrophication and fish kills. However, the development of environmental toxicology and chemistry has made it clear that more subtle and potential long-term effects also have to be taken into account in the regulatory work. The Swedish Environment Protection Board has recently completed a research project: "Characterization of Industrial Effluents." Initially, analytical methods and laboratory tests, originally developed for single substances, were identified. Laboratories were then invited to participate, with their own methods, in ring tests with a variety of industrial effluent waters. The studies indicated that a number of chemical and biological methods can be used to characterize complex industrial wastewaters with regard to their content of toxic, persistent, and bioaccumulative substances. A frame program was established for the construction of test batteries adapted to various administrative regulatory purposes. To date, this strategy has been applied to several industries by the authorities, resulting in valuable experience of cost-effectiveness and the usefulness in the administrative decision-making process. Besides the studies on single industries, two other approaches have been applied, namely for the assessment of the impact of several industries on a common receiving water body, and for the evaluation of a whole industrial branch, that is, kraft pulp mill bleachery effluents. In the latter case, the studies have resulted in a ranking of processes and treatment methods to protect the environment.

A simple method for collecting 14CO2 in fish.

A simple apparatus was designed for continuous collection of excreted 14CO2 in fish. Carp (Cyprinus carpio; body wt. about 400 g) were intraarterially injected with [14C]acetate (44 mmol) and transferred to a test chamber (21 +/- 1 degrees C). Within 48 h, 65.4 +/- 2.7% of the dose was recovered as 14CO2. A small amount of 14CO2 was retained in the water of the test chamber. Control experiments using [14C]acetate-injected mice as '14CO2-generators' showed that this 14C residue in the water could be recovered almost completely as 14CO2 upon acidification. In the carp experiments, some radioactivity remained in the acidified test chamber water; this residue may represent unchanged [14C]acetate and/or metabolites other than 14CO2.

Chlorinated paraffins: disposition of a polychloro-[1-14C]-hexadecane in carp (Cyprinus carpio) and bleak (Alburnus alburnus).

The disposition of polychloro-[1-14C]hexadecane (PCHD, 34% chlorine w/w) was studied in carp (Cyprinus carpio) and bleak (Alburnus alburnus). After intra-arterial (i.a.) injection in carp about 6% of dose was excreted as 14CO2 in 96 h. The autoradiographic distribution pattern after administration of PCHD to carp (i.a.) and bleak (up to 14 days of exposure in PCHD-contaminated water) was characterized by a strong labelling in the bile/intestinal contents. High radioactivity also appeared in the kidney, liver, gills and, particularly in bleak, in the nasal cavity, lens and skin.

Biological variables, especially skeletal deformities in fish, for monitoring marine pollution.

When considering principles for selection of indicators, i.e. biological variables, for monitoring marine pollution, it must be regarded as important to search for effects on the highest possible level of organization. For a global monitoring programme there are, however, many practical limitations in the number of useful indicators. The paper suggests skeletal deformities in fish as one possible indicator for this purpose in the future and gives a review on the occurrence, effects, causative factors and possible mechanisms of skeletal deformities in fish.

Automated thermoluminescence reader. I. Technical construction and function.

An automated TLD reader has been developed and constructed. The dosimeter material consisting of LiF is permanently enclosed in thin-walled teflon tubes. Dosimeters are identified by numbers, which simplifies individual calibration. A calibration radiation source consisting of a cylinder (90Sr-90Y) is used for the calibrations. The function of the reader and the calibration radiation source is described, and the method of measurement briefly reported.