Abstract
For aquatic systems, there is little data on the interactions between viruses, prokaryotes, grazers and the availability of resources. We conducted a microcosm experiment using a size fractionation approach to manipulate grazers, with a purpose to examine the effects of inorganic and organic nutrients on viral and prokaryotic standing stocks and activities, and on prokaryotic community composition as assessed by fluorescent in situ hybridization (FISH) method. Experiments were performed during periods of severe phosphate (P)-limiting conditions in the oligotrophic Sep Reservoir (Massif Central, France). In the absence of nutrient addition, the presence of grazers in microcosms stimulated prokaryotic growth and viral proliferation, likely through nutrient and substrate enrichment. Addition of nutrients had a stronger effect on viral infection of prokaryotes than grazing. Addition of P led to the most pronounced increase in prokaryotic abundance, production and growth efficiency, thus providing direct evidence of P limitation of prokaryotes. Enhanced prokaryotic activity in P treatments also stimulated viral abundance and viral-induced lyses of prokaryotes. Changes in prokaryotic community composition due to nutrient additions were evident in the grazer-free samples. Prokaryotic populations hybridizing for the probes bacteria, β-Proteobacteria and α-Proteobacteria responded to nutrient enrichment with significant increases in their relative abundances, whereas cells hybridizing for Archaea and Cytophaga-Flavobacterium (now known as Bacteroidetes) probes failed to show any functional response. Cells hybridizing for the latter cluster increased towards the end of incubation period in the control samples (that is, without nutrient additions) with grazers present, suggesting the development of grazing resistant forms. From our nutrient enrichment microcosm experiments, we conclude that the presence of grazers is a stimulating factor for prokaryotic growth and viral proliferation in the plankton, probably through nutrient regeneration process.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Amann R, Binder BJ, Olson RJ, Chisholm SW, Devereux R, Stahl DA . (1990). Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analysing mixed microbial populations. Appl Environ Microbiol 56: 1919–1925.
APHA (1985). Standard Methods for the Examination of Water and Wastewater, 16th edn. American Public Health Association: Washington, DC.
Arndt H, Dietrich D, Auer B, Cleven EJ, Grafenhan T, Weitere M et al. (2000). Functional diversity of heterotrophic flagellates in aquatic ecosystem. In: Leadbeater BSC, Green JC (eds). The Flagellates. Taylor and Francis Publishers: London, pp 240–268.
Becks L, Hilker FM, Malchow H, Jürgens K, Arndt H . (2005). Experimental demonstration of chaos in a microbial food web. Nature 435: 1226–1229.
Bettarel Y, Sime-Ngando T, Bouvy M, Arfi R, Amblard C . (2005). Low consumption of virus-sized particles by heterotrophic nanoflagellates in two lakes of the French Massif Central. Aquat Microb Ecol 39: 205–209.
Binder B . (1999). Reconsidering the relationship between viral induced bacterial mortality and frequency of infected cells. Aquat Microb Ecol 18: 207–215.
Felip M, Pace ML, Cole JJ . (1996). Regulation of planktonic bacteria growth rates: the effects of temperature and resources. Microb Ecol 31: 15–28.
Fuhrman JA . (1999). Marine viruses and their biogeochemical and ecological effects. Nature 399: 541–548.
Fuhrman JA, Azam F . (1982). Thymidine incorporation as a measure of heterotrophic bacterioplankton production in marine surface waters evaluation and field results. Mar Biol 66: 109–120.
Fuhrman JA, Noble RT . (1995). Viruses and protists cause similar bacterial mortality in coastal water. Limnol Oceanogr 40: 1236–1242.
Glöckner FO, Amann R, Alfreider A, Pernthaler J, Psenner R, Trebesius K et al. (1996). An in-situ hybridization protocol for detection and identification of planktonic bacteria. Syst Appl Microbiol 19: 403–406.
Glöckner FO, Fuchs BM, Amann R . (1999). Bacterioplankton compositions of lakes and oceans: A first comparison based on fluorescence in situ hybridization. Appl Environ Microbiol 65: 3721–3726.
Glöckner FO, Zaichikov E, Belkova N, Denissova L, Pernthaler J, Pernthaler A et al. (2000). Comparative 16SrRNA analysis of lake bacterioplankton reveals globally distributed phylogenetic clusters including an abundant group of Actinobacteria. Appl Environ Microbiol 66: 5053–5065.
Gonzàlez JM, Suttle CA . (1993). Grazing by marine nanoflaglellates on viruses and virus-sized particles: ingestion and digestion. Mar Ecol Prog Ser 94: 1–10.
Hornàk K, Masin M, Jezbera J, Bettarel Y, Nedoma J, Sime-Ngando T et al. (2005). Effects of decreased resource availability, protozoan grazing and viral impact on the structure of bacterioplankton assemblage in a canyon-shaped reservoir. FEMS Microbiol Ecol 52: 315–327.
Jacquet S, Domaizon I, Personnic S, Sime-Ngando T . (2007). Do small grazers influence virus-induced mortality of bacteria in Lake Bourget (France)? Fundam Appl Limnol 170: 125–132.
Jardillier L, Basset M, Domaizon I, Belan A, Amblard C, Richardot M et al. (2004). Bottom-up and top-down control of bacterial community composition in the euphotic zone of a reservoir. Aquat Microb Ecol 35: 259–273.
Jardillier L, Boucher D, Personnic S, Jacquet S, Thenot A, Sargos D et al. (2005). Relative importance of nutrients and mortality factors on prokaryotic community composition in two lakes of different trophic status: microcosm experiments. FEMS Microbiol Ecol 53: 429–443.
Jugnia JB, Tadonléké RD, Sime-Ngando T, Devaux J . (2000). The microbial food web in the recently flooded reservoir: diel fluctuations in bacterial biomass and metabolic activity in relation to phytoplankton and flagellate grazers. Microb Ecol 40: 317–329.
Jugnia JB, Tadonléké RD, Sime-Ngando T, Devaux J, Andrivon C . (1999). Bacterial population dynamics, production and heterotrophic activity in a recently formed reservoir. Can J Microbiol 45: 747–753.
Jürgens K, Pernthaler J, Schalla S, Amann R . (2002). Morphological and compositional changes in a planktonic bacterial community in response to enhanced protozoan grazing. Appl Environ Microbiol 65: 1241–1250.
Kirchman DL . (2002). The ecology of Cytophaga-Flavobacteria in aquatic environments. FEMS Microbiol Ecol 39: 91–100.
Kooi BW, Boer MP . (2003). Chaotic behaviour of a predator-prey system in the chemostat. Dynam Discrete Impulsive Syst B 10: 259–272.
Ludwig W, Klenk HP . (2001). Overview: a phylogenetic backbone and taxonomic framework for prokaryotic systematics. In: Boone DR, Castenholz RW (eds). Bergy's Manual of Systematic Bacteriology. Springer-Verlag: New York, pp 49–65.
Manz W, Amann R, Ludwig W, Wagner M, Schleifer KH . (1992). Phylogenic oligodeoxynucleotide probes for the major subclasses of proteobacteria: problems and solutions. Syst Appl Microbiol 15: 593–600.
Middelboe M, Jorgensen NOG, Kroer N . (1996). Effects of viruses on nutrient turnover and growth efficiency of non-infected marine bacterioplankton. Appl Environ Microbiol 62: 1991–1997.
Noble RT, Fuhrman J . (1999). Breakdown and microbial uptake of marine viruses and other lysis products. Aquat Microb Ecol 20: 1–11.
Noble RT, Fuhrman JA . (1998). Use of SYBR Green I for rapid epifluorescence counts of marine viruses and bacteria. Aquat Microb Ecol 14: 113–118.
Pernthaler J, Glockner FO, Unterholzner S, Alfreider A, Psenner R, Amann R . (1998). Seasonal community and populations dynamics of pelagic bacteria and Archaea in a high mountain lake. Appl Environ Microbiol 64: 4299–4306.
Pradeep Ram AS, Boucher D, Sime-Ngando T, Debroas D, Romagoux JC . (2005). Phage bacteriolysis, protistan bacterivory potential, and bacterial production in a freshwater reservoir: coupling with temperature. Microb Ecol 50: 64–72.
Pradeep Ram AS, Nair S, Chandramohan D . (2003). Bacterial growth efficiency in the tropical estuarine and coastal waters of Goa, southwest coast of India. Microb Ecol 45: 88–96.
Pradeep Ram AS, Nair S, Chandramohan D . (2007). Bacterial growth efficiency in a tropical estuary: seasonal variability subsidized by allochthonous carbon. Microb Ecol 53: 591–599.
Richardot M, Debroas D, Thouvenot A, Sargos D, Berthon JL, Dévaux J . (2001). Influence of cladoceran grazing activity on dissolved organic matter, enzymatic hydrolysis and bacterial growth. J Plankton Res 23: 1249–1261.
Schwalbach MS, Hewson I, Fuhrman JA . (2004). Viral effects on bacterial community in a marine plankton microcosms. Aquat Microb Ecol 34: 117–127.
SCOR-UNESCO (1966). Determination of Photosynthetic Pigments in Sea Water. UNESCO: Paris, pp 1–69.
Šimek K, Hornàk K, Jezbera J, Nedoma J, Vrba J, Straskràbovà V et al. (2006). Maximum growth rates and possible life strategies of different bacterioplankton groups in relation to phosphorus availability in a freshwater reservoir. Environ Microbiol 8: 1613–1624.
Šimek K, Hornàk K, Masin M, Christaki U, Nedoma J, Weinbauer MG et al. (2003). Comparing the effects of resource enrichment and grazing on a bacterioplankton community of a meso-eutrophic reservoir. Aquat Microb Ecol 31: 123–135.
Šimek K, Pernthaler J, Weinbauer MG, Hornàk K, Dolan J, Nedoma J et al. (2001). Changes in bacterial community composition and dynamics and viral mortality rates associated with enhanced flagellate grazing in mesotrophic reservoir. Appl Environ Microbiol 67: 2723–2733.
Sime-Ngando T, Bettarel Y, Chartogne C, Sean K . (2003). The imprint of wild viruses on freshwater microbial ecology. Recent Res Dev Microbiol 7: 481–497.
Sime-Ngando T, Mignot JP, Amblard C, Bourdier G, Devilettes C, Quilblier-Lloberas C . (1996). Characterization of planktonic virus-like particles in the French mountain lake: methodological aspects and preliminary results. Ann Limnol 32: 1–5.
Sime-Ngando T, Pradeep Ram AS . (2005). Grazer effects on prokaryotes and viruses in a freshwater microcosm experiment. Aquatic Microb Ecol 41: 115–124.
Stahl DA, Amann R . (1991). Development and application of nucleic acid probes. In: Stackebrandt E, Goodfellow M (eds). Nucleic Acid Techniques in Bacterial Systematics. John Wiley: Chichester, UK, pp 205–248.
Suttle CA . (2007). Marine viruses—major players in the global ecosystem. Nat Rev Microbiol 5: 801–812.
Tadonléké DR, Jugnia LB, Sime-Ngando T, Devaux J, Romagoux JC . (2002). Food web structure in the recently flooded Sep Reservoir as inferred from phytoplankton dynamics and living microbial biomass. Microb Ecol 43: 67–81.
Tadonléké DR, Sime-Ngando T, Amblard C, Sargos D, Devaux J . (2000). Primary productivity in a recently flooded reservoir. J Plankton Res 21: 179–199.
Thingstad TF, Lignell R . (1997). Theoretical models for the control of bacterial growth rate, abundance, diversity and carbon demand. Aquat Microb Ecol 13: 19–27.
Vaynes DV, Pavlou S . (1999). Chaotic dynamics of a food web in a chemostat. Math Biosci 162: 69–84.
Vrede K . (2005). Nutrient and temperature limitation of bacterioplankton growth in temperate lakes. Microb Ecol 49: 245–256.
Weinbauer MG . (2004). Ecology of prokaryotic viruses. FEMS Microbiol Rev 28: 127–181.
Weinbauer MG, Christaki U, Nedoma J, Šimek K . (2003). Comparing the effects of resource enrichment and grazing on viral production in a mesoeutrophic reservoir. Aquat Microb Ecol 31: 137–144.
Weinbauer MG, Hornàk K, Jezbera J, Nedoma J, Dolan JR, Šimek K . (2007). Synergistic and antagonistic effects of viral lysis and protistan grazing on bacterial biomass, production and diversity. Environ Microbiol 9: 777–788.
Weinbauer MG, Winter C, Höfle MG . (2002). Reconsidering transmission electron microscopy based estimates of viral infection of bacterioplankton using conversion factors derived from natural communities. Aquat Microb Ecol 27: 103–110.
Weiss P, Schweitwer B, Amann R, Simon M . (1996). Identification in situ and dynamics of bacteria on limnetic organic aggregates (lake snow). Appl Environ Microbiol 62: 1998–2005.
Wilhelm SW, Suttle CA . (1999). Viruses and nutrient cycles in the sea. Bioscience 49: 781–788.
Williamson SJ, Paul JH . (2004). Nutrient stimulation of lytic phage production in bacterial populations of the Gulf of Mexico. Aquat Microb Ecol 36: 9–17.
Wilson W, Joint I, Carr NG, Mann NH . (1996). The effect of phosphorus status on the kinetics of cyanophage infection in the oceanic cyanobacterium Synechoccus sp. WH7803. J Phycol 32: 506–516.
Wilson WH, Mann NH . (1997). Lysogenic and lytic viral production in marine microbial communities. Aquat Microb Ecol 13: 95–100.
Acknowledgements
We thank M Weinbauer for his constructive comments and suggestions on the paper. ASPR was supported by a postdoctoral fellowship from the French Government (Ministry of Youth, National Education and Research). The study was partly supported by the French national program ACI/FNS ‘ECCO’ (VIRULAC research grant awarded to TSN). We thank C Portelli, D Boucher, D Debroas and A Thenot for their logistic, technical and field assistance.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Pradeep Ram, A., Sime-Ngando, T. Functional responses of prokaryotes and viruses to grazer effects and nutrient additions in freshwater microcosms. ISME J 2, 498–509 (2008). https://doi.org/10.1038/ismej.2008.15
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/ismej.2008.15
Keywords
This article is cited by
-
Distribution patterns of bacterial communities and their potential link to variable viral lysis in temperate freshwater reservoirs
Aquatic Sciences (2019)
-
Taxonomical Resolution and Distribution of Bacterioplankton Along the Vertical Gradient Reveals Pronounced Spatiotemporal Patterns in Contrasted Temperate Freshwater Lakes
Microbial Ecology (2018)
-
Contribution of Viral Lysis and Nanoflagellate Grazing to Bacterial Mortality at Surface Waters and Deeper Depths in the Coastal Ecosystem of Subtropical Western Pacific
Estuaries and Coasts (2016)
-
The microbial aspect of climate change
Energy, Ecology and Environment (2016)
-
Viral lysis and nanoflagellate grazing on prokaryotes: effects of short-term warming in a coastal subtropical marine system
Hydrobiologia (2015)