The cultivation and growth behavior of metal-tolerant strains of Streptomyce acidiscabies E13 and Streptomyces sp. F4 were studied under droplet-based microfluidics conditions. It was shown that the technique of micro segmented flow is well suited for the investigation of dependence of bacterial growth on different concentrations of either single metal ions or combinations of them. This study confirms higher tolerance to Zn than to Cu by our test organism. The highly resolved dose–response curves reflect two transitions between the different growth behaviors, separating initial responses to Cu concentration ranges into those with (a) intense growth, (b) moderate growth, and (c) growth inhibition. For Streptomyces sp. F4, an initial stimulation was shown in the sublethal range of zinc sulfate. Two-dimensional screenings using computer-controlled fluid actuation and in situ micro flow-through fluorimetry reflected a strong growth stimulation of strain F4 by zinc sulfate in the presence of sublethal Cu concentrations. This stimulatory effect on binary mixtures may be useful in providing optimal growth conditions in bioremediation procedures.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Admiraal W, Blanck H, Buckert-De Jong M, Guasch H, Ivorra N, Lehmann V, Nystrom BAH, Paulsson M, Sabater S (1999) Short-term toxicity of zinc to microbenthic algae and bacteria in a metal polluted stream. Water Res 33:1989–1996
Amoroso MJ, Schubert D, Mitscherlich P, Schumann P, Kothe E (2000) Evidence for high affinity nickel transporter genes in heavy metal resistant Streptomyces spec. J Basic Microb 40:295–301
Barbulovic-Nad I, Yang H, Park PS, Wheeler AR (2008) Digital microfluidics for cell-based assays. Lab Chip 8:519–526
Blinova I, Ivask A, Heinlaan M, Mortimer M, Kahru A (2010) Ecotoxicity of nanoparticles of CuO and ZnO in natural water. Environ Pollut 158:41–47
Cao JL, Kursten D, Schneider S, Knauer A, Gunther PM, Kohler JM (2012) Uncovering toxicological complexity by multi-dimensional screenings in microsegmented flow: modulation of antibiotic interference by nanoparticles. Lab Chip 12:474–484
Churski K, Kaminski TS, Jakiela S, Kamysz W, Baranska-Rybak W, Weibel DB, Garstecki P (2012) Rapid screening of antibiotic toxicity in an automated microdroplet system. Lab Chip 12:1629–1637
Clausell-Tormos J, Merten CA (2012) Micro segmented-flow in biochemical and cell-based assays. Front Biosci 4:1768–1779 (Elite Ed)
Costa JSD, Kothe E, Abate CM, Amoroso MJ (2012) Unraveling the Amycolatopsis tucumanensis copper-resistome. Biometals 25:905–917
Franklin NM, Stauber JL, Lim RP, Petocz P (2002) Toxicity of metal mixtures to a tropical freshwater alga (Chlorella sp.): The effect of interactions between copper, cadmium, and zinc on metal cell binding and uptake. Environ Toxicol Chem 21:2412–2422
Funfak A, Brosing A, Brand M, Kohler JM (2007) Micro fluid segment technique for screening and development studies on Danio rerio embryos. Lab Chip 7:1132–1138
Funfak A, Hartung R, Cao JL, Martin K, Wiesmuller KH, Wolfbeis OS, Kohler JM (2009) Highly resolved dose–response functions for drug-modulated bacteria cultivation obtained by fluorometric and photometric flow-through sensing in microsegmented flow. Sensor Actuat B-Chem 142:66–72
Funfak A, Cao JL, Knauer A, Martin K, Kohler JM (2011) Synergistic effects of metal nanoparticles and a phenolic uncoupler using microdroplet-based two-dimensional approach. J Environ Monitor 13:410–415
Gherbal R, Hamed H, Foth H (2011) The effect of zinc ions on expression of metallothionein and poly (ADP-ribose) polymerase-1 in human lung cells. Toxicol Lett 205:S160
Gunther PM, Schneider S, Groß GA, Kohler JM (2011) Addressing of multidimensional concentration spaces by micro segmented flow technique. Proc Mikrosyst Technol CongrDarmst 10:945–947
Haferburg G, Kothe E (2010) Metallomics: lessons for metalliferous soil remediation. Appl Microbiol Biot 87:1271–1280
Ince NH, Dirilgen N, Apikyan IG, Tezcanli G, Ustun B (1999) Assessment of toxic interactions of heavy metals in binary mixtures: a statistical approach. Arch Environ Con Tox 36:365–372
Kalantari N, Ghaffari S (2008) Evaluation of toxicity of heavy metals for Escherichia coli growth. Iran J Environ Health 5:173–178
Kohler JM, Henkel T, Grodrian A, Kirner T, Roth M, Martin K, Metze J (2004) Digital reaction technology by micro segmented flow—components, concepts and applications. Chem Eng J 101:201–216
Kuersten D, Cao J, Funfak A, Mueller P, Köhler JM (2011) Cultivation of Chlorella vulgaris in microfluid segments and microtoxicological determination of their sensitivity against CuCl2 in the nanoliter range. Eng Life Sci 11:1–8
Le TTY, Vijver MG, Hendriks AJ, Peijnenburg WJGM (2013) Modeling toxicity of binary metal mixtures (Cu2+–Ag+, Cu2+–Zn2+) to lettuce, Lactuca sativa, with the biotic ligand model. Environ Toxicol Chem 32:137–143
Malik A (2004) Metal bioremediation through growing cells. Environ Int 30:261–278
Martin K, Henkel T, Baier V, Grodrian A, Schon T, Roth M, Kohler JM, Metze J (2003) Generation of larger numbers of separated microbial populations by cultivation in segmented-flow microdevices. Lab Chip 3:202–207
Navarro E, Baun A, Behra R, Hartmann NB, Filser J, Miao AJ, Quigg A, Santschi PH, Sigg L (2008) Environmental behavior and ecotoxicity of engineered nanoparticles to algae, plants, and fungi. Ecotoxicology 17:372–386
Nies AT, Damme K, Schaeffeler E, Schwab M (2012) Multidrug and toxin extrusion proteins as transporters of antimicrobial drugs. Expert Opin Drug Met 8:1565–1577
Rademacher C, Masepohl B (2012) Copper-responsive gene regulation in bacteria. Microbiol-Uk 158:2451–2464
Sani RK, Peyton BM, Brown LT (2001) Copper-induced inhibition of growth of Desulfovibrio desulfuricans G20: assessment of its toxicity and correlation with those of zinc and lead. Appl Environ Microb 67:4765–4772
Schmidt A, Haferburg G, Sineriz M, Merten D, Buchel G, Kothe E (2005) Heavy metal resistance mechanisms in actinobacteria for survival in AMD contaminated soils. Chem Erde-Geochem 65:131–144
Schmidt A, Schmidt A, Haferburg G, Kothe E (2007) Superoxide dismutases of heavy metal resistant streptomycetes. J Basic Microb 47:56–62
Schmidt A, Haferburg G, Schmidt A, Lischke U, Merten D, Ghergel F, Buchel G, Kothe E (2009) Heavy metal resistance to the extreme: Streptomyces strains from a former uranium mining area. Chem Erde-Geochem 69:35–44
Schmidt A, Rzanny M, Schmidt A, Hagen M, Schutze E, Kothe E (2012) GC content-independent amino acid patterns in Bacteria and Archaea. J Basic Microb 52:195–205
Theberge AB, Courtois F, Schaerli Y, Fischlechner M, Abell C, Hollfelder F, Huck WTS (2010) Microdroplets in microfluidics: an evolving platform for discoveries in chemistry and biology. Angew Chem Int Edit 49:5846–5868
Utgikar VP, Tabak HH, Haines JR, Govind R (2003) Quantification of toxic and inhibitory impact of copper and zinc on mixed cultures of sulfate-reducing bacteria. Biotechnol Bioeng 82:306–312
Valko M, Morris H, Cronin MTD (2005) Metals, toxicity and oxidative stress. Curr Med Chem 12:1161–1208
Zheng B, Tice JD, Ismagilov RF (2004) Formation of droplets of alternating composition in microfluidic channels and applications to indexing of concentrations in droplet-based assays. Anal Chem 76:4977–4982
The financial support of the German Federal Ministry of Education and Research (BMBF) in the frame of the project “BactoCat” (Kz: 031A161A) is gratefully acknowledged. J. Cao is financially supported by the German Federal Environmental Foundation.
About this article
Cite this article
Cao, J., Kürsten, D., Krause, K. et al. Application of micro-segmented flow for two-dimensional characterization of the combinatorial effect of zinc and copper ions on metal-tolerant Streptomyces strains. Appl Microbiol Biotechnol 97, 8923–8930 (2013). https://doi.org/10.1007/s00253-013-5147-8
- Droplet-based microfluidics
- Segmented flow
- Heavy metal resistance
- Combinatorial effects