Abstract
Background, aim, and scope
Dissolved humic substances (HSs) are exogenous stressors to aquatic plants and animals which activate a variety of transcriptional and biochemical reactions or block photosynthesis. While there are consistent indications which structures may lead to reduced photosynthetic activity, there is much less clear information available on which HS structures or building blocks act as stressors in animals. Consequently, this work was designed to comparatively study the impact of natural organic matter (NOMs) from different sources on major anti-stress mechanisms in one single animal. We utilized major antioxidant responses and relative expression levels of stress proteins (small HSPs and HSP70) and expected that different HSs provoke different response patterns.
Materials and methods
We tested the freshwater amphipod Gammarus pulex which was collected from several shallow creeks in Northern Germany. All specimens were maintained in aerated 5-L tanks with filtered water from their natural environment at 10°C with prior acclimation. Animals were fed ad libitum with a commercial preparation once every second day. The exposure water was exchanged with the same frequency. NOMs were isolated from three different sources: two from small brown-water lakes in Northern Germany by reverse osmosis and the third one as an aqueous extract from a black layer of a Brazilian sandbar soil (State of Rio de Janeiro). The rationale was to apply NOMs of contrasting quality. Chemical fingerprint features of the NOMs were taken by high-performance size exclusion chromatography. As stress parameters in the animals, the activities of peroxidase and catalase were recorded quantitatively, and stress proteins, HSP70, as well as small α-crystalline HSPs were analyzed semiquantitatively.
Results
The three NOMs clearly differed in molecular masses, humic substance contents, the moieties of polysaccharides, and low-molecular-weight substances. With the exception of one short-term response, the peroxidase activity increased after 3 to 12 h exposure, whereas the catalase activity did not show any significant modulation. With one exception, the stress protein expression increased after 30 min exposure in a biphasic pattern, and the sHSPs responded less strongly than HSP70.
Discussion
Although the quality of the exposed NOMs differs significantly, a rather uniform response pattern appears in the animals. Obviously, the contrasting contents of HSs and polysaccharides did not affect the anti-stress response of the exposed gammarids which is in contrast to previous lifespan studies with Caenorhabditis elegans. Furthermore, all NOM sources led to increased contents of both HSP70 and sHSPs. To the best of our knowledge, this is the first protein study to show that also small HSPs are expressed when the animals are exposed against humic material.
Conclusions
Since the response patterns of the exposed gammarids, in contrast to the initial hypothesis, are rather uniform and since HSs are parts of life on Earth, we furthermore presume that they may have been a primordial exogenous trigger for the development of anti-stress systems in exposed organisms.
Recommendations and perspectives
Effect studies of chemical stresses on organisms should consider exposure to both natural triggers and xenobiotic compounds in low concentrations—in order to prospectively differentiate between these triggers and, subsequently, classify them.
Similar content being viewed by others
References
Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126
Arrigo A (1998) Small stress proteins: chaperones that act as regulators of intracellular redox state and programmed cell death. Biol Chem 379:19–26
Bergmeyer HU (1983) Methods of enzymatic analysis, vol I. VCH Weinheim, Germany
Blokhina O, Violainen E, Fagerstedt KV (2003) Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Bot 91:179–194
Bradford MM (1976) Rapid and sensitive method for quantification of microgram quantities of protein utilizing principle of protein dye binding. Anal Biochem 72:248–254
Cazenave J, de Ángeles Bistoni M, Zwirnmann E, Wunderlin DA, Wiegand C (2006) Attenuating effects of natural organic matter on microcystin toxicity in zebra fish (Danio rerio) embryos—benefits and costs of microcystin detoxication. Environ Toxicol 21:22–32
DIN EN 1484 (1998) Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlammuntersuchung: Anleitung zur Bestimmung des gesamten organischen Kohlenstoffs (TOC) und des gelösten organischen Kohlenstoffs (DOC). Wiley, Weinheim
Dröge W (2002) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95
Elayan NM, Treleaven WD, Cook RL (2008) Monitoring the effect of three humic acids on a model membrane system using P NMR. Environ Sci Technol 42:1531–1536
Euent S, Menzel R, Steinberg CEW (2008) Gender-specific lifespan modulation in Daphnia magna by a dissolved humic substances preparation. Ann Environ Sci 2:7–10
Farjalla VF, Faria BM, Esteves FA (2002) The relationship between DOC and planktonic bacteria in tropical coastal lagoons. Arch Hydrobiol 156:97–119
Farjalla VF, Enrich-Prast A, Esteves FA, Cimbleris ACP (2006) Bacterial growth and DOC consumption in a tropical coastal lagoon. Braz J Biol 66:383–392
Fent K (2007) Ökotoxikologie: Umweltchemie, Toxikologie, Ökologie. Thieme, Stuttgart
Galvez F, Donini A, Playle RC, Smith DS, O'Donnell MJ, Wood CM (2008) A matter of potential concern: natural organic matter alters the electrical properties of fish gills. Environ Sci Technol 42:9385–9390
Gjessing ET, Alberts JJ, Bruchet A, Egeberg PK, Lydersen E, McGown LB, Mobed JJ, Münster U, Pempkowika J, Perdue M, Ratnawerra H, Rybacki D, Takacs M, Abbt-Braun G (1998) Multi-method characterisation of natural organic matter isolated from water: characterisation of reverse osmosis-isolates from water of two semi-identical dystrophic lakes basins in Norway. Wat Res 32:3108–3124
Karasyova TA, Klose EO, Menzel R, Steinberg CEW (2007) Natural organic matter differently modulates growth of two closely related coccal green algal species. Environ Sci Pollut Res 14:88–93
Kley A, Maier G (2005) An example of niche partitioning between Dikerogammarus villosus and other invasive and native gammarids: a field study. J Limnol 64:85–88
Kley A, Maier G (2006) Reproductive characteristics of invasive gammarids in the Rhine-Main-Danube catchment, South Germany. Limnologica 36:79–90
Kolesnichenko A, Zykova V, Grabelnych O, Sumina O, Pobezhimova T, Voinikov V (1999) Screening of mitochondrial proteins in winter rye, winter wheat, elymus and maize with an immunochemical affinity to the stress protein 310 kD and their intramitochondrial localisation in winter wheat. J Therm Biol 24:290–296
Laemmli U (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature 227:680–685
Lowry O, Rosebrough N, Farr A, Randall R (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Mason CF (1981) Biology of freshwater pollution. Longman, London
Matsuo AYO, Woodin BR, Reddy CM, Val AL, Stegeman JJ (2006) Humic substances and crude oil induce cytochrome P450 1A expression in the Amazonian fish species Colossoma macropomum (tambaqui). Environ Sci Technol 40:2851–2858
Meems N, Steinberg CEW, Wiegand C (2004) Humic substances and the insecticide cypermethrin: direct and indirect effects on the waterflea (Daphnia magna). Sci Total Environ 319:123–136
Meinelt T, Phan TM, Zwirnmann E, Krüger A, Paul A, Wienke A, Steinberg CEW (2007) Reduction in vegetative growth of the water mold Saprolegnia parasitica (Coker) by humic substances of different origin. Aquat Toxicol 83:93–103
Menzel R, Stürzenbaum S, Kulas J, Bärenwaldt A, Steinberg CEW (2005) Humic material induces behavioral and global transcriptional responses in the nematode Caenorhabditis elegans. Environ Sci Technol 39:8324–8332
Miller SL (1955) Production of some organic compounds under possible primitive earth conditions. J Am Chem Soc 77:2351–2361
Münster U (1985) Investigation about structure, distribution and dynamics of different organic substrates in the DOM of lake Plusssee. Arch Hydrobiol/Suppl 70:429–480
Papp E, Nardai G, Soti C, Csermely P (2003) Molecular chaperones, stress proteins and redox homeostasis. Biofactors 17:249–257
Paul A, Pflugmacher S, Steinberg CEW (2003) Correlation of spin concentration in humic substances with inhibitory effects on photosynthesis of aquatic macrophytes. Akad Wissensch Erfurt, Sitzber Mathem Naturw Kl 12:209–221 (in German)
Paul A, Hackbarth S, Vogt RD, Röder B, Burnison BK, Steinberg CEW (2004) Photogeneration of singlet oxygen by humic substances: comparison of humic substances of aquatic and terrestrial origin. Photochem Photobiol Sci 3:272–280
Pflugmacher S, Pietsch C, Rieger W, Steinberg CEW (2006) Influence of structural moieties of dissolved organic matter on the photosynthetic oxygen production of aquatic plants. Sci Total Environ 357:169–175
Prokhotskaya VYu, Steinberg CEW (2007) Differential sensitivity of a coccal green algal and a cyanobacterial species to dissolved natural organic matter (NOM). Environ Sci Pollut Res 14(SI 1):11–18
Rudolph K (2001) Die Flohkrebsfauna (Crustacea, Amphipoda) der Länder Brandenburg und Berlin. Natursch Landschaftspfl Brandenburg 10:166–172
Sachse A, Babenzien D, Ginzel G, Gelbrecht J, Steinberg CEW (2001) Characterization of dissolved organic carbon (DOC) of a dystrophic lake and an adjacent fen. Biogeochemistry 54:279–296
Steinberg CEW (2003) Ecology humic substances in freshwaters—determinants from geochemistry to ecological niches. Springer, Heidelberg
Steinberg CEW, Münster U (1985) Geochemistry and ecological role of humic substances in lakewater. In: Aiken GR, McKnight DM, Wershaw RL, MacCarthy P (eds) Humic substances in soil, sediment, and water. Wiley, New York, pp 105–145
Steinberg CEW, Paul A, Pflugmacher P, Meinelt T, Klöcking R, Wiegand C (2003) Pure humic substances have the potential to act as xenobiotic chemicals—a review. Fresenius Environ Bull 12:391–401
Steinberg CEW, Manusadžianas L, Grigutytė R, Karitonas R, Jurkoniene S, Pflugmacher S (2004) Membrane depolarisation and elevation of ROS-defensive mechanisms due to the impact of dissolved natural organic matter (NOM) in the Charophyte Nitellopsis obtusa. In: Martin-Neto L, Milori DMBP, da Silva WTL (eds) Humic substances and soil and water environment. Embrapa, São Carlos, pp 135–137
Steinberg CEW, Kamara S, VYu P, Manusadžianas L, Karasyova T, Timofeyev MA, Zhang J, Paul A, Meinelt T, Farjalla VF, Matsuo AYO, Burnison BK, Menzel R (2006) Dissolved humic substances—ecological driving forces from the individual to the ecosystem level? Freshw Biol 51:1189–1210
Steinberg CEW, Saul N, Pietsch K, Meinelt T, Rienau S, Menzel R (2007) Dissolved humic substances facilitate fish life in extreme aquatic environments and have the potential to extend lifespan of Caenorhabditis elegans. Ann Environ Sci 1:81–90
Steinberg CEW, Meinelt T, Timofeyev MA, Bittner M, Dissolved MR (2008) Dissolved humic substances (review series). Part 2: interactions with organisms. Env Sci Pollut Res 15:128–135
Suhett AL, Amado AM, Enrich-Prast A, Esteves FDA, Farjalla VF (2007) Seasonal changes of dissolved organic carbon photo-oxidation rates in a tropical humic lagoon: the role of rainfall as a major regulator. Can J Fish Aquat Sci 64:1266–1272
Thurman EM (1985) Aquatic humic substances. Organic geochemistry of natural waters. Martinus Nijhoff/Dr W. Junk, Dordrecht
Timofeyev MA, Steinberg CEW (2006) Comparative study of the antioxidant response to natural organic matter (NOM) exposure in three Baikalean amphipod species from contrasting habitats. Comp Biochem Physiol B 145:197–203
Timofeyev M, Wiegand C, Burnison BK, Shatilina ZM, Pflugmacher S, Steinberg CEW (2004) Impact of natural organic matter (NOM) on freshwater amphipods). Sci Total Environ 319:115–121
Timofeyev MA, Shatilina ZM, Kolesnichenko AV, Kolesnichenko VV, Pflugmacher S, Steinberg CEW (2006a) Natural organic matter (NOM) promotes oxidative stress in freshwater amphipods Gammarus lacustris Sars and G. tigrinus (Sexton). Sci Total Environ 366:673–681
Timofeyev MA, Shatilina ZM, Kolesnichenko AV, Kolesnichenko VV, Steinberg CEW (2006b) Specific antioxidants reaction to oxidative stress promoted by natural organic matter (NOM) in two endemic amphipod species from Lake Baikal. Environ Toxicol 21:104–110
Timofeyev M, Shatilina ZM, Bedulina DS, Menzel R, Steinberg CEW (2007) Natural organic matter (NOM) has the potential to modify the multixenobiotic resistance (MXR) activity in freshwater amphipods Eulimnogammarus cyaneus (Dyb) and Eulimnogammarus verrucosus (Gerst.). Comp Biochem Physiol B 146:496–503
Towbin H, Staehelin T, Gordon T (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Nat Acad Sci 76:4350–4354
Vigneault B, Percot A, Lafleur M, Campbell PGC (2000) Permeability changes in model and phytoplankton membranes in the presence of aquatic humic substances. Environ Sci Technol 34:3907–3913
Visser SA (1985) Physiological action of humic substances on microbial cells. Soil Biol Biochem 17:457–462
Vogt RD, Akkanen J, Andersen DO, Brüggemann R, Gjessing E, Kukkonen J, Luster J, Paul A, Pflugmacher S, Starr M, Steinberg CEW, Schmitt-Kopplin P, Zsolnay A (2004) Key site variables governing the functional characteristics of dissolved (DNOM) in Nordic forested catchments. Aquat Sci 66:195–210
Wetzel RG (2001) Limnology. Lake and river ecosystems. Academic, San Diego
Wiegand C, Pflugmacher S, Oberemm A, Meems N, Beattie KA, Steinberg CEW, Codd GA (1999) Uptake and effects of microcystin-LR on detoxication enzymes of early life stages of the zebra fish Danio rerio. Environ Toxicol 14:89–95
Ziechmann W (1994) Humic substances. BI-Wissenschaftsverlag, Mannheim
Ziechmann W (1996) Huminstoffe und ihre Wirkungen. Spektrum Akademischer Verlag, Heidelberg
Zink KG, Furtado ALS, Casper P, Schwark L (2004) Organic matter composition in the sediment of three Brazilian coastal lagoons—District of Macaé, Rio de Janeiro (Brazil). Ann Braz Acad Sci 76:29–47
Acknowledgments
This research was partially supported by grants of Russian Foundation for Basic Research No 06-04-48099-а, 08-04-00928-а, 08-04-10065-a, RosObrazovanie No. 2.1.1/982 and joint Russian Ministry of education and DAAD grant program “Mikhail Lomonosov”. The supports are gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Additional information
Responsible editors: Christian E. W. Steinberg and Markus Hecker
Rights and permissions
About this article
Cite this article
Bedulina, D.S., Timofeyev, M.A., Zimmer, M. et al. Different natural organic matter isolates cause similar stress response patterns in the freshwater amphipod, Gammarus pulex . Environ Sci Pollut Res 17, 261–269 (2010). https://doi.org/10.1007/s11356-009-0222-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11356-009-0222-5