Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

Left atrial rupture in dogs: 14 cases (1990–2005)

Objective: To describe the clinical parameters, treatment, and prognosis of dogs with left atrial rupture secondary to chronic mitral valve insufficiency. Design: Retrospective study. Setting: University referral hospital. Animals: 14 dogs with left atrial rupture. Interventions: None. Measurements and main results: Mixed breed dogs (n=6, 43%) and Shetland Sheepdogs (n=3, 21%) were most commonly affected. The median age was 12 years (range 5.8–18 y). The median weight was 11 kg (range 4–30 kg). Eight dogs had been previously diagnosed with chronic valvular disease. The most common presenting complaints included collapse (13/14), cough (9/14), and dyspnea (8/14). Four dogs were presented in either respiratory or cardiac arrest. Pericardial effusion was present in 13 dogs. The median left atrium:aortic outflow ratio was 2.66 (range 1.66:1–5.52:1). Pericardiocentesis was performed to alleviate tamponade in 3 dogs. Five dogs were discharged from the hospital, 3 of which were euthanized within 35 days of initial diagnosis for recurrence of clinical signs (n=2) and for hematochezia and lethargy (n=1). Five dogs were euthanized while in the hospital for a variety of reasons including DIC, progressive azotemia, collapse and recurrence of pericardial effusion, or possible seizure episode. Conclusions: Although rare, left atrial rupture resulting in pericardial effusion should be considered in older small‐ to medium‐sized dogs presenting with collapse, cough, and dyspnea. The overall prognosis appears poor.     

Association of holidays, full moon, Friday the 13th, day of week, time of day, day of week, and time of year on case distribution in an urban referral small animal emergency clinic

Objective – To interrogate the association of variables: day of week, time of day, day of the year (major holidays, Friday the 13th, and the full moon), and month of year with the caseload of an urban academic emergency service.
Design – Retrospective study.
Setting – Urban small animal teaching hospital emergency clinic.
Animals – Cats and dogs that were presented to the emergency clinic.
Interventions – None.
Measurements and Main Results – The hospital computer database was searched for all visits to the Emergency Service of the Mathew J. Ryan Veterinary Hospital of the University of Pennsylvania from January 1, 1987 through December 31, 2002. Variables included in the electronic data were date of admission, time of day of admission, species (dog or cat), hospital service the case was transferred to for ongoing care and whether the case was discharged directly from the emergency service. The association of caseload with day of week, time of day, day of the year (major holidays, Friday the 13th, and the full moon), and month of year was described and statistically evaluated. Saturdays and Sundays were the busiest days of the week and significantly increased caseload was noted for the majority of holidays (except Easter Day and Thanksgiving Day) with Memorial Day being the busiest. Midweek evenings as well as Saturday and Sunday afternoons were the busiest periods of the day. There was no association with caseload and Halloween, the full moon, or Friday the 13th.
Conclusions – The busiest times were midweek evenings, Saturday and Sunday afternoons, and most major holidays.     

Rapid emergence of baculovirus resistance in codling moth due to dominant, sex-linked inheritance

Insect-specific baculoviruses are increasingly used as biological control agents of lepidopteran pests in agriculture and forestry, and they have been previously regarded as robust to resistance development by the insects. However, in more than a dozen cases of field resistance of the codling moth Cydia pomonella to commercially applied C. pomonella granulovirus (CpGV) in German orchards, resistance ratios exceed 1000. The rapid emergence of resistance is facilitated by sex-linkage and concentration-dependent dominance of the major resistance gene and genetic uniformity of the virus. When the gene is fixed, resistance levels approach 100,000-fold. Our findings highlight the need for development of resistance management strategies for baculoviruses.