Dirk Jungmann

Stefan Scholz; Erika Sela; Ludek Blaha; Thomas Braunbeck; Malyka Galay-Burgos; Mauricio García-Franco; Joaquin Guinea; Nils Klüver; Kristin Schirmer; Katrin Tanneberger; Marysia Tobor-Kapłon; Hilda Witters; Scott E. Belanger; Emilio Benfenati; Stuart Creton; Mark T. D. Cronin; Rik I. L. Eggen; Michelle R. Embry; Drew R. Ekman; Anne Gourmelon; Marlies Halder; Barry Hardy; Thomas Hartung; Bruno Hubesch; Dirk Jungmann; Mark A. Lampi; Lucy E. J. Lee; Marc Léonard; Eberhard Küster; Adam Lillicrap

Tests with vertebrates are an integral part of environmental hazard identification and risk assessment of chemicals, plant protection products, pharmaceuticals, biocides, feed additives and effluents. These tests raise ethical and economic concerns and are considered as inappropriate for assessing all of the substances and effluents that require regulatory testing. Hence, there is a strong demand for replacement, reduction and refinement strategies and methods. However, until now alternative approaches have only rarely been used in regulatory settings. This review provides an overview on current regulations of chemicals and the requirements for animal tests in environmental hazard and risk assessment. It aims to highlight the potential areas for alternative approaches in environmental hazard identification and risk assessment. Perspectives and limitations of alternative approaches to animal tests using vertebrates in environmental toxicology, i.e. mainly fish and amphibians, are discussed. Free access to existing (proprietary) animal test data, availability of validated alternative methods and a practical implementation of conceptual approaches such as the Adverse Outcome Pathways and Integrated Testing Strategies were identified as major requirements towards the successful development and implementation of alternative approaches. Although this article focusses on European regulations, its considerations and conclusions are of global relevance.

Dirk Jungmann; Vanessa Ladewig; Kai-Uwe Ludwichowski; Patrick Petzsch; Roland Nagel

The occurrence of intersexuality was investigated in Gammarus fossarum populations from five different streams in Germany in the years 2000-2003. In the streams Lockwitzbach and Korsch sampling sites were upstream and downstream of sewage treatment plant effluents, in the three other streams an effluent was absent. In the stream Lockwitzbach the median frequency of intersexes at the two sampling sites was 7.1 % and 13 % over two years. In the streams Korsch and Zschonerbach the frequency of intersexes was lower with 0.2 % and 0.8 % at the two sampling sites at the Korsch, respectively, and between 0.3 % and 0.7% at the Zschonerbach. Because in the Goldersbach and the Friedrichsbach in the absence of effluents the frequency of intersexes was 3.2% and 24%, an impact of chemicals from municipal sewage treatment plants can be excluded. As the phenomenon occurred in three of five streams but can easily be overlooked, it may be much more common than previously assumed. At the upstream sampling site of the Lockwitzbach with a high frequency of intersex the affected specimens were larger compared to females. An exposure of gammarids from streams with low intersex frequency to water from streams with high intersex frequency led to a significant, time dependent increase of intersex frequency. Thus, although there are reports about induction of intersex by photoperiod, a factor which plays no role in the experiments performed here, it is concluded that some factor in the water - may be a chemical - is responsible for the induction of intersex in G. fossarum. At the moment it is unclear whether an anthropogenic or natural factor is responsible and which role environmental conditions play.

Vanessa Ladewig; Dirk Jungmann; Anna Koehler; M. Schirling; Rita Triebskorn; Roland Nagel

We present the first description of intersexes in Gammarus fossarum Koch, 1835 (Amphipoda). Intersexes were found in monthly samples collected in two streams in different regions of Germany. The frequency of intersexuality differed considerably between the populations from the two streams. The Lockwitzbach, located in Sachsen, showed a frequency of 8.8% intersexes, which is much higher compared to the K?rsch (located in Baden-W?rttemberg) with 0.6%. In contrast to the K?rsch, there were also differences between the two sampling sites in the Lockwitzbach. No seasonal pattern in intersex frequency was observed. Intersexes occurred in all length classes of the adolescent and adult gammarids examined. They were significantly larger than females, but showed no difference in body length compared to males. External sex characteristics of intersex individuals have been described in detail, while in histological analyses of internal sex characteristics ovarian tissue was found exclusively. Microsporidians, parasites often associated with intersexuality, were not detected. The intersexes seemed to be functional females, regarding their ability to breed.

Dirk Jungmann; Kristin Brust; Oliver Licht; Jens Mählmann; Jens Schmidt; Roland Nagel

The potential hazard of chemicals on aquatic communities are generally evaluated by standardised single-species bioassays. Safety assessment is based on results gained from organisms adapted to lentic systems and biological interactions in ecosystems are neglected. While lotic communities are often at first in contact with chemicals, it is astonishing that microcosms with lentic communities are mainly used as a bridge between laboratory bioassays and outdoor aquatic systems. Hence, we established five artificial indoor streams to simulate abiotic factors of small rivers. The closed-circuit system was filled with nutrients added to tap water. Washed pebbles were used as sediment. The dynamics of a simple biocoenoses consisting of aufwuchs,Lumbriculus variegatus Asellus aquaticus andGammarus fossarum was investigated. The dynamic of aufwuchs and periphyton was determined as dry weight and chlorophyll-a, respectively and qualitatively by pigment pattern. The abundance of different developmental stages ofL. variegatus was determined at the end of the experiment as well as the population dynamics ofG. fossarum andA. aquaticus. Survival rates of gammarids and juveniles per female were investigated and data were used for modelling the population dynamics. The experiment was carried out to investigate the performance of the established artificial streams and the developed approaches to investigate effects of chemicals on a basic lotic community. The prime reason to establish this approach was to close a gap between complex artificial stream systems and laboratory single species tests to assess the impact of chemicals on the aquatic environment.

Rita Triebskorn; Kristin Berg; Ina Ebert; Manfred Frey; Dirk Jungmann; Jörg Oehlmann; Matthias Oetken; Frank Sacher; Marco Scheurer; Hannah Schmieg; Simon Schwarz; Heinz-R. Köhler

Environment with Mode of Action-Specific in Vitro Biotests Rita Triebskorn,*,†,‡ Kristin Berg, Ina Ebert, Manfred Frey, Dirk Jungmann, Jörg Oehlmann, Matthias Oetken, Frank Sacher, Marco Scheurer, Hannah Schmieg,† Simon Schwarz,† and Heinz-R. Köhler† †Animal Physiological Ecology, University of Tübingen, Auf der Morgenstelle 5, D-72076 Tübingen, Germany ‡Steinbeis Transfer Center for Ecotoxicology and Ecophysiology, Blumenstr. 13, D-72108 Rottenburg, Germany GWT-Technical University Dresden, Blasewitzer Str. 43, D-01307 Dresden, Germany Federal Environment Agency, Wörlitzer Platz 1, D-06844 Dessau-Roßlau, Germany Steinbeis Innovation Center Cell Culture Technology, Schulzenstr. 4, D-68259 Mannheim, Germany Aquatic Ecotoxicology, University of Frankfurt am Main, Max-von-Laue-Str. 13, D-60438 Frankfurt, Germany DVGW Water Technology Center, Karlsruher Str. 84, D-76139 Karlsruhe, Germany

Hai-Jun Wang; Xucheng Xiao; Hongzhu Wang; Yan Li; Qing Yu; Xiao-Min Liang; Weisong Feng; Jian-Chun Shao; Marcus Rybicki; Dirk Jungmann; Erik Jeppesen

A number of studies have revealed ammonia to be toxic to aquatic organisms; however, little is known about its effects under natural conditions. To elucidate the role of ammonia, we conducted 96-h acute toxicity tests as well as a whole-ecosystem chronic toxicity test for one year in ten 600-m2 ponds. Three common cyprinids, silver carp Hypophthalmichthys molitrix Val. (H.m.), bighead carp Aristichthys nobilis Richardson (A.n.), and gibel carp Carassius auratus gibelio Bloch (C.g.), were used as test organisms. The 96-h LC50 values of un-ionized ammonia (NH3) for H.m., A.n., and C.g. were 0.35, 0.33, and 0.73mgL-1, respectively. In the ponds, annual mean NH3 ranged between 0.01 and 0.54mgL-1, with 4 ponds having a NH3 higher than the LC50 of A.n. (lowest LC50 in this study). No fish were found dead in the high-nitrogen ponds, but marked histological changes were found in livers and gills. Despite these changes, the specific growth rate of H.m. and A.n. increased significantly with NH3. Our pond results suggest that fish might be more tolerant to high ammonia concentrations in natural aquatic ecosystems than under laboratory conditions. Our finding from field experiments thus suggests that the existing regulatory limits for reactive nitrogen (NH3) established from lab toxicity tests might be somewhat too high at the ecosystem conditions. Field-scale chronic toxicity tests covering full life histories of fish and other aquatic organisms are therefore encouraged in order to optimize determination of the effects of ammonia in natural environments.

Dirk Jungmann; Kristin Berg; Andreas Dieterich; Martin Frank; Tonya Gräf; Marco Scheurer; Simon Schwarz; Carmen Siewert; Matthias Oetken

ABSTRACT The aim of this study was to determine the effect data for metoprolol as a model substance for beta-blockers in aquatic invertebrates. The results will be used as a basis for the validation of future mode of action-based in vitro test systems targeting this class of pharmaceuticals. Effects of metoprolol were investigated in two autochthonous species with high relevance in stream ecology: the amphipod Gammarus fossarum and the oligochaete Lumbriculus variegatus. Mortality in G. fossarum was not observed in acute toxicity testing (48 h), and a significant increase of mortality at 45 mg/L was found when amphipods were exposed chronically (40 days). The most sensitive population-relevant endpoints were the juvenile-adult ratio and number of egg-bearing females with NOEC/LOEC-values of 5/15 mg/L. No proteotoxic effects were identified in G. fossarum. The sediment toxicity test with L. variegatus according to the OECD Guideline 225 with an exposure time of 28 days resulted in EC10-values of 92.5 and 126.1 mg/kgdw for the endpoints reproduction and biomass, respectively. In L. variegatus the response kinetics of Hsp70 showed no significant difference between the treatments. A tendency for rising lipid peroxide concentrations was found between 0.03 and 10 mg/kgdw, which were significant between the treatments, but not to the control.

Rico Illing; Corinna Burkart; Daniel Pfitzner; Dirk Jungmann; Larysa Baraban; Gianaurelio Cuniberti

Precise analysis of the aquatic cells and their responses to the toxic chemicals, i.e., water disinfective agents, is of crucial importance due to their role in the ecosystem. We demonstrate the application of the droplets based millifluidic tool for isolating and longtime monitoring of single Paramecium tetraurelia cells using a large number of water-in-oil emulsion droplets. Due to the automated monitoring of the fluorescence signal, the droplets containing cells are distinguished from the empty reservoirs. A viability indicator is used to follow the metabolic dynamic of the cells in every single droplet. Finally, we perform ecotoxicity tests in droplets, exposing the encapsulated paramecia cells to silver nitrate for determination of EC50 levels, and compare the output with the conventional microtiter plate assay.

Vanessa Ladewig; Dirk Jungmann; H.-R. Köhler; Oliver Licht; Kai-Uwe Ludwichowski; M. Schirling; Rita Triebskorn; Roland Nagel

The impact of bisphenol A (BPA) on Gammarus fossarum and Lumbriculus variegatus was studied in four artificial indoor streams (0, 5, 50 and 500 µg L−1 BPA, nominal) over 103 days in a pulse–dose exposure scenario (weekly BPA application). For G. fossarum populations at day 103, the proportions of juveniles and of breeding females from the highest BPA treatment were in tendency reduced. For individually exposed gammarid pairs an EC10 of 17 µg L−1 BPA (nominal) for the proportion of reproductive females in the fourth brood was determined. During the first three broods, the largest brood size occurred at the highest BPA concentration, whereas in the fourth brood it decreased concentration-dependently (fourth brood EC10 = 5 µg L−1 BPA, nominal). Effects on L. variegatus were a reduced population growth (103 d-EC10 of 2 µg L−1 BPA, nominal) and an increase in dry weight and the number of segments in large, complete worms.

Anna-Katharina Buchberger; Denise Brettschneider; Kristin Berg; Dirk Jungmann; Jörg Oehlmann; Marco Scheurer; Matthias Oetken

ABSTRACT Aquatic organisms are impacted by various biotic and abiotic stressors such as current, inter- and intraspecific competition for food resources and habitat, neobiota as well as an increasing number of chemicals. The latter also include pharmaceuticals, which are increasingly being detected in surface waters due to their growing use. The aim of our study was to determine effect data for metoprolol as a model compound for beta-blockers under an environmentally realistic exposure scenario on aquatic invertebrates inhabiting lotic environments. To this end we performed a 40-day experiment in artificial indoor streams (AIS) located in a greenhouse. We focussed on three autochthonous invertebrate species with high relevance in stream ecology: the amphipod Gammarus fossarum, the gastropod Potamopyrgus antipodarum, and the oligochaete Lumbriculus variegatus. Effects on reproduction were found with EC10 (40 days) values of 0.092 mg L−1 (G. fossarum), 0.253 mg L−1 (P. antipodarum), and 0.596 mg L−1 (L. variegatus). Considering environmental data, metoprolol seems to pose no hazard for aquatic invertebrates at present exposure levels.