A farewell to Dr Thomas Knacker (* 29.04.1951 – ? 30.10.2011): scientific contributions and personal memories

Background

The bioaccumulation potential of chemical substances is commonly determined in flow-through fish tests according to the Organization for Economic Cooperation and Development (OECD) technical guideline (TG) 305, aiming at the determination of bioconcentration factors (BCF). Investigations on aquatic organisms have shown a significant positive correlation between the accumulation of a chemical and the lipid content of organisms. Residue levels can thus differ between individuals, species, and size groups depending on their lipid content. BCF values can be normalized by lipid content; however, this requires the use of appropriate extraction techniques to ensure the complete extraction of total lipids from fish/tissue samples. The OECD TG 305 is currently under revision. A literature review was carried out to select preferred methods which shall be included in the updated OECD TG 305 along with a sampling schedule for lipid measurement in fish.

Results

An overview and appraisal of a broad range of lipid extraction methods is presented. Extraction procedures applied in bioaccumulation studies described in the literature and procedures currently used for such studies by governmental, academic, and industrial labs are surveyed. Possible impacts of unsuitable extraction techniques on lipid normalized BCF results were assessed. Default values for BCF normalization commonly used in bioaccumulation studies are described. The effect of differences in residue distribution over different organs and/or lipids on the normalization of BCF values is discussed.

Conclusions

A broad range of procedures for lipid extraction are available. However, only the use of suitable extraction techniques guarantees the complete extraction of total lipids from fish samples in bioaccumulation studies which is required to ensure the correct lipid normalization of BCF values. Preferred methods for inclusion in the updated OECD 305 test guideline as well as a standard schedule for lipid measurements are suggested.     

Fishery Enhancement Reef Building Exercise

The constructive use of coal-fired power station waste products for building artificial reefs is being explored for the first time in the U.K. at present, the practice of sea dumping of raw pulverised fuel ash (PFA) from coal combustion is under review and the use of consolidated PFA blocks offers a possible alternative. the planned fitting of flue gas desulphurisation (FGD) plant in the U.K. will additionally produce large quantities of gypsum. to test the environmental compatability of such materials in marine structures, an experimental reef has been constructed off the central south coast of the U.K. Fifty tonnes of blocks made from different combinations of PFA, gypsum, FGD waste water sludge, cement and gravel using standard concrete as the control material, were deployed. the site selected is remote from prominent sea bed features in an area of flat sand with limited species variety.The preliminary studies leading to the licensing of this project included bioassay experiments with diatom cultures, field trials with test tiles and trace metal bioaccumulation studies the stages of consultation and licensing of the project are outlined.

Colonisation of the reef has been rapid, initially by shoaling fish, particularly by bib (Trisopterus luscus) then, within the first month, by representatives of the commercial crab and lobster species typical of the local natural rocky coast. After three months the epifauna and flora were well established. at this stage some 80 species of flora and fauna have been identified in association with the reef with no differentiation in colonisation apparent between the PFA/gypsum mixtures and the concrete control. the number of species far exceeds that of PFA powder dumping grounds. Analyses for heavy metal ions in the reef blocks after 2 months immersion have not shown any significant difference from the original composition.     

Effect of Chlorination on the Gill Lipids of the Mussel Mytilus edulis (L)

Experiments are described which involve the chlorination of standard solutions of lipids and populations of the mussel, Mytilus edulis (L). The object was to test the hypothesis that chlorine will bind to unsaturated lipids. Evidence is presented that chlorine and aqueous chlorinated species rapidly and readily form addition compounds with unsaturated lipids in vivo as well as in vitro. The relevance of these findings to possible environmental effects of chlorination is discussed.     

Modelling the effect of acidity on mercury uptake by walleye in acidic and circumneutral lakes

An association between acidification of lakes and high mercury concentrations in fishes has been reported. In northern Wisconsin (USA), for example, walleye (Stizostedion vitreum) in naturally acidic lakes have higher mercury concentrations than walleye in circumneutral lakes (Weiner, 1983). In this study a bioenergetic based model for mercury uptake is applied to identify potential causes of higher mercury concentrations in fishes living in acidic lakes. Application of the model to walleye populations in northern Wisconsin indicates that higher mercury concentrations in acidic lakes are most likely a result of both increased concentrations in the water and increased efficiencies of uptake from water. Higher concentrations in the water would result in higher concentrations in the food.     

Effect of tetracycline residues in pig manure slurry on tetracycline-resistant bacteria and resistance gene tet(M) in soil microcosms

Effects of tetracycline residues from pig manure slurry on the prevalence of tetracycline-resistant bacteria and the tetracycline resistance gene, tet(M), were studied in soil microcosms. Four types of soil microcosms were established for a period of 152 days, supplemented with combinations of pig manure slurry and a tetracycline-resistant Enterococcus faecalis, CG110, containing the tetracycline resistance gene tet(M) (on the conjugative transposon, Tn916). The prevalence of both tetracycline-resistant aerobic bacteria and tetracycline-resistant enterococci declined rapidly until day 45 where no significant differences in the levels of tetracycline-resistant bacteria in any of the four types of microcosms could be detected. tet(M) could be detected in microcosms supplemented with either pig manure slurry and/or E. faecalis CG110 (tet(M)) for the whole period (152 days). tet(M) could be detected longer than tetracycline-resistant enterococci could be isolated (limit of detection 100 CFU/g soil) probably due to viable but not culturable (VBNC) bacteria with tet(M), horizontal gene transfer of tet(M) to indigenous soil bacteria or presence of "free" DNA. The concentration of chlortetracycline and oxytetracycline were almost stable through out the experimental period, but the tetracycline concentrations had no effect on prevalence of tetracycline-resistant bacteria. The presented microcosm approach simulated natural farmland conditions well and supported results from previous field studies.     

Influence of plant growth on degradation of linear alkylbenzene sulfonate in sludge-amended soil

Widespread application of sewage sludge to agricultural soils in Denmark has led to concern about the possible accumulation and effects of linear alkylbenzene sulfonate (LAS) in the soil ecosystem. Therefore, we have studied the uptake and degradation of LAS in greenhouse pot experiments. Sewage sludge was incorporated into a sandy soil to give a range from very low to very high applications (0.4 to 90 Mg dry wt. ha(-1)). In addition, LAS was added as water solutions. The soil was transferred to pots and sown with barley (Hordeum vulgare L. cv. Apex), rape (Brassica napus L. cv. Hyola 401), or carrot (Daucus carota L.). Also, plant-free controls were established. For all additions there was no plant uptake above the detection limit at 0.5 mg LAS kg(-1) d.w, but plant growth stimulated the degradation. With a growth period of 30 d, LAS concentrations in soil from pots with rape had dropped from 27 to 1.4 mg kg(-1) dry wt., but in plant-free pots the concentration decreased only to 2.4 mg kg(-1) dry wt. When LAS was added as a spike, the final concentration in soil from planted pots was 0.7 mg kg(-1) dry wt., but in pots without plants the final concentration was much higher (2.5 mg kg(-1) dry wt.). During degradation, the relative fraction of homologues C10, C11, and C12 decreased, while C13 increased.     

Determination of selected pharmaceuticals and caffeine in sewage and seawater from Tromsø/Norway with emphasis on ibuprofen and its metabolites

Selected pharmaceuticals, among them analgesics, ss-blockers and anti-depressants as well as caffeine, the anti-bacterial triclosan and the insect repellent N,N-diethyl-3-toluamide (DEET) were determined in different sewage samples (sewage treatment plants, hospital effluents) from Troms?/Norway and in seawater from Troms?-Sound, into which the sewage is discharged. While caffeine, triclosan, ibuprofen and its major metabolites hydroxy- and carboxy-ibuprofen were present in all sewage samples, additional pharmaceuticals were observed in sewage containing hospital effluents. Concentrations were in the range of 20-293 microg/l (caffeine), 0.2-2.4 microg/l (triclosan) and 0.1-20 microg/l (sum ibuprofen + metabolites). In seawater, only caffeine (7-87 ng/l), DEET (0.4-13 ng/l) and ibuprofen + metabolites (sum concentration < LOQ-7.7 ng/l) were detected. Ibuprofen and its metabolites hydroxy- and carboxy-ibuprofen were quantified individually by use of the respective reference compounds. Relative amounts of the three compounds were determined in different types of water showing characteristic patterns, with hydroxy-ibuprofen being the major component in sewage whereas carboxy-ibuprofen was dominant in seawater samples. The patterns which were compared to those observed in similar samples from Germany indicated different transformation behaviour under limnic and marine conditions.     

Status and trends of fecal indicator bacteria in two urban watersheds.

This paper examines bacterial levels and their causes in two Houston bayous (Texas). Buffalo and Whiteoak bayous are two of the most contaminated water bodies in Texas for indicator bacteria, based on the frequency and magnitude of contact recreation water quality exceedances. Examination of historical data indicates frequent exceedances, although some improvement has been made since the 1970s. Statistical analyses showed some correlation between in-stream fecal coliform concentrations and rainfall and with land use. Differences in fecal coliform concentrations were found between high- and low-flow conditions in Whiteoak Bayou, while reservoir releases confounded this relationship in Buffalo Bayou. Wastewater treatment plant effluent was found to make up two-thirds to three-fourths of the median flow in both bayous. Effluent sampling was conducted at 72 of the approximately 140 wastewater treatment plants (WWTPs) in the watersheds, providing evidence that WWTP effluent could act to maintain low-flow concentrations of fecal coliform in the bayous.