This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Colombet, J.
Right arrow Articles by Demeure, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Colombet, J.
Right arrow Articles by Demeure, G.
Agricola
Right arrow Articles by Colombet, J.
Right arrow Articles by Demeure, G.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, June 2006, p. 4440-4445, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.00021-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Depth-Related Gradients of Viral Activity in Lake Pavin

J. Colombet,1 T. Sime-Ngando,1* H. M. Cauchie,2 G. Fonty,1 L. Hoffmann,2 and G. Demeure1

Laboratoire de Biologie des Protistes, Université Blaise Pascal (Clermont-Ferrand II), UMR CNRS 6023, F-63177 Aubière Cedex, France,1 CRP-Gabriel Lippmann, Cellule de Recherche en Environnement et Biotechnologies, 162a Avenue de la Faïencerie, L-1511 Luxembourg, Luxembourg2

Received 4 January 2006/ Accepted 14 March 2006


arrow
ABSTRACT
 
High-resolution vertical sampling and determination of viral and prokaryotic parameters in a deep volcanic lake shows that in the absence of thermal stratification but within light, oxygen, and chlorophyll gradients, host availability empirically is prevalent over the physical and chemical environments and favors lytic over lysogenic "viral life cycles."


arrow
INTRODUCTION
 
Viral activity is crucial to microbial mortality, diversity, potential gene transfer, and lysogenic conversion in aquatic systems (see references 18 and 19 for recent reviews). Viruses can affect ecological processes and nutrient cycles (23), but this ultimately depends on whether the virus is lytic or temperate. Studies examining large data sets have revealed that prokaryotes form the major host reservoir for viruses in pelagic systems (2). Protistan grazing and viral lysis are major sources of bacterial mortality in these systems. In general, bacterivory dominates, but losses of bacteria from viral lysis at times are comparable to those from protistan bacterivory (13). In conditions such as anoxia, bacteriolysis generally is much higher than bacterivory based on evidence from theoretical (12) and empirical (2) field investigations. In contrast to bacterivory and lytic viral infection, which have been examined in a variety of marine systems, only two studies have investigated the two viral "life cycles" simultaneously, but not together with bacterivory (11, 21). For freshwater, one study has examined temperate phages in the surface layer of Lake Superior (17).

Pelagic viral ecology thus lacks studies in which potential prophage induction, lytic viral infection, and bacterivory are investigated simultaneously. In addition, most of the studies on these processes have ignored the well-known contribution of depth-related gradients as a major forcing factor in aquatic systems. Inclusion of such a contribution is needed to fully assess the potential roles of viruses for microbial food web processes and to address hypotheses that (i) in suboxic and anoxic waters the importance of lytic viral infection increases relative to bacterivory (2, 13, 20) and (ii) lysogeny increases as lytic viruses decrease with their host densities and production (21). Here we address these questions by examining the vertical distribution of the frequencies of lytically and lysogenically infected cells together with the potential bacterivory from heterotrophic nanoflagellates in a deep meromictic lake. Samples were collected in early spring, when the isothermal water column showed gradients of light, oxygen, and chlorophyll, in order to avoid false interpretations of correlations that often result from the covariation of two variables with respect to a third factor, usually temperature.

Lake Pavin is a deep (Zmax = 92 m) meromictic mountain lake located in the French Massif Central (2°56'E, 45°29'N). Samples were taken in triplicate on 20 April 2004 at the reference station (located in the middle and the deepest part of the lake), using an 8-liter Van Dorn bottle. A total of 15 depths were sampled in three distinct layers, comprising 7 depths in the mixolimnion (0.5, 5, 15, 20, 30, 40, and 50 m), 5 depths in the oxycline, (56, 57, 57.5, 58, and 59 m), and 3 depths in the anoxic monimolimnion (60, 70, and 80 m). Water temperature and dissolved oxygen were measured in situ with an Oxycal-SL 197 multiparameter probe (WWT, Limonest, France). Secchi depth (Zs) measurements were used to estimate the euphotic depth (Zeu) according to the relationship Zeu = 2.42Zs (22). Chlorophyll a concentrations were determined spectrophotometrically following extraction in 90% acetone (16). For determination of viral and bacterial abundances, subsamples were fixed with 0.02-µm-filtered buffered alkaline formalin (final concentration, 2% [vol/vol]) immediately after sampling and filtered (<15-kPa vacuum) through 0.02-µm-pore-size Anodisc filters (Whatman, Maidstone, United Kingdom). Counts were done under a Leica DC 300F epifluorescence microscope (Episcope) following staining with SYBR green I fluorochrome (Molecular Probes Europe, Leiden, The Netherlands) as described by Noble and Fuhrman (10). When not analyzed immediately, slides were stored at –20°C until counting was performed.

Bacterial production was determined by the incorporation of [3H]leucine (final concentration, 40 nM; specific activity, 71 Ci mmol–1) (Amersham Biosciences, United Kingdom) into bacterial biomass by using the microcentrifuge method (9). The incubation time (30 min in situ in the dark) was fixed from preliminary experiments conducted on 25 March 2004, and protein precipitation in controls (i.e., organisms killed with 5% trichloroacetic acid) and fixed assays was aided by treatment with NaCl (final concentration, 3.5% [vol/vol]) at 18°C for 30 min. After centrifuge washings, microbial pellets were dissolved in 0.2 ml 1.2 N NaOH at 80°C for 20 min, and scintillation cocktail (1 ml) (Ready Safe; Beckman Coulter) was added for radioactivity counting with a Beckman LS 6500 liquid scintillation counter. For suboxic and anoxic samples, incubations were done in 5-ml sterile serum bottles sealed with rubber and aluminum crimp caps and flushed with N2. Leucine incorporation was converted into the number of cells produced by using conversion factors (0.27 x 1018 cells mol–1 for oxic waters and 0.25 x 1018 cells mol–1 for suboxic and anoxic waters) determined during preliminary experiments conducted on 25 March 2004.

For viral lytic infection, the frequency of infected cells (FIC) and the virus-induced bacterial mortality were calculated from the frequency of visibly infected cells obtained from observations under a JEOL 1200EX transmission electron microscope, following ultracentrifugation and uranyl acetate staining. The procedure is detailed elsewhere (2, 13). For each sample, mean burst size was estimated from the number of viruses in those infected cells which were filled with phages. In addition, free viruses were examined for the relative abundances of three arbitrary divided size classes based on the capsid diameter: <60, 60 to 100, and >100 nm. Viral lytic production was estimated by multiplying bacterial cell production by FIC/100 and the burst size. The frequency of lysogenically infected cells (FLC) was determined from the induction of prophages by using mitomycin C (8). Mitomycin C was added to samples (final concentration, 1 µg ml–1) in 20-ml sterile serum bottles, and untreated samples served as controls. For the suboxic and anoxic samples, serum bottles were sealed with rubber and aluminum crimp caps and flushed with N2. The incubation time (24 h in situ) was fixed from a 76-hour time series preliminary experiment conducted on 25 March 2004. Subsamples were removed with syringes at 0 and 24 h and fixed with glutaraldehyde for viral and bacterial counts. FLC was estimated from viral abundances in mitomycin C-treated (VAm) and control (VAc) incubations and bacterial abundance (BAt0) and burst size (BSt0) in original samples, as follows: FLC = 100[(VAm – VAc)/(BSt0 x BAt0)] (21).

Samples for enumeration of heterotrophic nanoflagellates (HNF) were fixed with glutaraldehyde (final concentration, 1%) immediately after sampling. Primulin-stained HNF collected on 0.8-µm polycarbonate black filters were counted under UV excitation using the LEICA Episcope (2, 13). Potential grazing rates of heterotrophic nanoflagellates were estimated as the product of bacterial concentration, flagellate concentration, and assumed mean clearance rate of 6.1 nl flagellate–1 h–1 (range, 0.7 to 11.5 nl flagellate–1 h–1) reported by Carrias et al. (5) for a seasonal study on Lake Pavin. These values fell within the range of values published for freshwater systems (15).

Differences between mixolimnion, oxycline, and monimolimnion were compared with standard analysis of variance. Potential relationships among variables were tested by Pearson correlation analysis. All data were analyzed after logarithmic transformations.


arrow
The vertical environment.
 
During the study, the isothermal water column temperature was at about the point of maximum density (4.3°C). The water transparency (Secchi depth = 6 m) represented half of the annual maximum (5), and the euphotic zone extended over one-third of the mixolimnion (22). Oxygen concentrations in the water column reflected the annual range, i.e., from anoxia to 12 mg liter–1 (2, 5). Based on these concentrations, the three layers of the water column were characterized as oxic (O2 concentration of >8 mg liter–1), suboxic (O2 concentration of between 0.5 and 8 mg liter–1), and anoxic (O2 concentration of between 0 and 0.5 mg liter–1), corresponding to mixolimnion (depth of 0 to 50 m), oxycline (50 to 59 m), and monimolimnion (60 to 92 m), respectively (Table 1; Fig. 1). Pigment concentrations (2 to 13 µg liter–1) also reflected the typical annual range and were related to the development of the large diatoms Aulacoseira italica and Asterionella formosa, which, during the sampling period, typically accounted for 50 to 98% of the total phytoplankton biomass in Lake Pavin. Deep chlorophyll a peaks in Lake Pavin, observed at depths of 40 and 57.5 m during this study, typically are known to be the result of the sedimentation of these diatoms (1).


View this table:
[in this window]
[in a new window]
  		TABLE 1. Characteristics of samples collected in the three main layers of Lake Pavin


Figure 1
View larger version (10K):
[in this window]
[in a new window]
  		FIG. 1. Vertical variations of oxygen and chlorophyll a concentrations in Lake Pavin. Error bars represent standard deviations for triplicates. The three layers (mixolimnion, oxycline, and deep monimolimnion) of the sampled water column are shown.


arrow
Viral size classes.
 
Phage polyhedral head sizes in our samples ranged from 10 to 130 nm, and tail sizes ranged from 10 to 180 nm. Tailed viruses accounted for 75% of the total free viruses, indicating that most of the viruses were bacteriophages. We considered this to be conservative because they may have been nontailed bacteriophages or tailed bacteriophages that lost their tails during ultracentrifugation or had tails with low contrast or no contrast during the staining procedure. Phages with head sizes of ≤60 nm clearly dominated the viral community in oxygenated waters (Fig. 2), corroborating reports from the world aquatic systems, where the size class of 30 to 60 nm is normally dominant within the virioplankton (19, 24). However, this generalization is weakened here, because the three viral size classes were about equally abundant in hypoxic and anoxic deep waters, suggesting that the composition of viral communities may differ markedly according to vertical gradients in aquatic systems. For example, eukaryotic (algae, metazooplankton, and fish) and allochthonous (from catchment and atmosphere) viruses may be more prevalent in the mixolimnion than in the monimolimnion of deep meromictic lakes such as Lake Pavin.


Figure 2
View larger version (15K):
[in this window]
[in a new window]
  		FIG. 2. Vertical variations of the relative abundances of viral size classes (i.e., percentage of total observed), of viral abundance (VA) and lytic production (VP), of bacterial abundance (BA) and production (BP), and of the abundance of HNF in Lake Pavin. Error bars represent standard deviations for triplicates. The three layers (mixolimnion, oxycline, and deep monimolimnion) of the sampled water column are shown.


arrow
Standing stocks.
 
Viral abundances (range 3 x 106 to 37 x 106 particles ml–1) peaked in the anoxic monimolimnion and were lower than the seasonal abundances (10 x 106 to 50 x 106 viruses ml–1) reported by Bettarel et al. (3) for the thermally stratified mixolimnion of Lake Pavin. Our bacterial abundances (7 x 106 to 25 x 106 cells ml–1) were up to one order of magnitude higher than those reported by Bettarel and coauthors (2 x 106 to 9 x 106 cells ml–1), due to the very high bacterial stock in the monimolimnion. Compared to the seasonal values (mean = 7.1 ± 1.9) in the stratified mixolimnion of Lake Pavin (3), our virus-to-bacterium ratios (VBRs) (mean = 2.3 ± 0.4) were lower but similarly varied little compared to abundances. In the Mediterranean and Baltic Seas, low bacterial stocks in suboxic and anoxic deep waters resulted in particularly high VBRs (up to 50) compared to that in the surface layer (21). The vertical distributions of viral and bacterial abundances during our study were similar and were negatively correlated with oxygen (Table 2; Fig. 2). The numerical values and spatial fluctuations in the upper oxic layer were low, followed by an exponential increase towards the suboxic and anoxic layers. Together with the low variability in VBR, this confirms previous indications that most of the free viruses in the plankton were bacteriophages and that there is a close coupling between viral and bacterial concentrations (2, 3). Similar vertical trends were reported for viruses in a small eutrophic lake (20) and for bacteria in a meromictic lake (7). Surprisingly high viral densities in mesopelagic and deep marine waters (depth of up to 2,000 m) were also reported (6, 21). For Lake Pavin, the possibility that monimolimnic viruses come from the surface layers is unlikely because of the chemocline barrier and the indication from the size class analysis that they were indigenous to the deep waters. It has been suggested that high abundances of free viruses in deep marine waters may be due to an increased survival rate of viruses in low-temperature waters because viral infectivity decreased with decreasing temperature (21). Our empirical data do not seem to support this hypothesis, at least for the meromictic Lake Pavin, which was isothermal during our study. It is thus likely that biological processes, including lytic activity and induction of prophages in conjunction with host availability, were prevalent over the physical environment in the viral proliferation in deep anoxic waters.


View this table:
[in this window]
[in a new window]
  		TABLE 2. Correlation relationships of basic parameters in the water column of Lake Pavin

HNF cell numbers in this study (0 to 4 x 103 cells ml–1) were at the lower end of the seasonal ranges (2 x 103 to 20 x 103 cells ml–1) in the thermally stratified mixolimnion (3) but were similar to those (0.8 x 103 to 2.2 x 103 cells ml–1) reported during early spring in the same lake (4). The spatial pattern of HNF abundances clearly contrasted with those of viral and bacterial abundances (Fig. 2). HNF peaked in the surface, decreased with depth, and were almost absent in the deep anoxic waters. This pattern was negatively correlated with viral and bacterial abundances and positively correlated with oxygen (Table 2), implying that deep anoxic conditions excluded typical bacterivores.


arrow
Lytic viral infection versus potential bacterivory.
 
Inclusion of the monimolimnion layer in our study (Table 1) resulted in higher FIC levels (7 to 79%) than the seasonal values (5 to 40%) reported for the stratified mixolimnion of Lake Pavin (2). In other aquatic systems, this variable generally is <50% (24). Bacterial lytic mortality in the monimolimnion was about two- and eightfold higher than those in the oxycline and mixolimnion, respectively, and almost all the mortality in the anoxic deep waters was through viral lysis (Fig. 3; Table 1). The same vertical pattern was observed for the calculated viral lytic production (Fig. 2). This implies that the high viral densities in the monimolimnion were produced in situ. The sharp contrast observed between the vertical patterns of viral bacteriolysis and potential HNF bacterivory supports the idea that in environments such as suboxic and anoxic waters, the importance of lytic viral infection increases relative to bacterivory (2, 13, 20). FIC and lytic production of viruses were indeed positively correlated with bacterial abundance and production but negatively correlated with oxygen and HNF abundance and potential grazing (Table 2). Increasing lytic mortality with depth has also been reported for the anoxic deep waters of the Baltic Sea, with values up to 71% (21), similar to those reported here for the monimolimnion of Lake Pavin. These values are among the highest reported for marine and freshwater systems (19, 24). We suggest that they are related to the low grazing pressure and to the high host abundance and production, perhaps of low diversity, in deep anoxic waters.


Figure 3
View larger version (11K):
[in this window]
[in a new window]
  		FIG. 3. Vertical variations of lytic virus-mediated (VBM) and potential grazing-mediated mortalities in bacterial assemblages and of the FIC and FLC in Lake Pavin. BP, bacterial production. Error bars represent standard deviations for triplicates. The three layers (mixolimnion, oxycline, and deep monimolimnion) of the sampled water column are shown.


arrow
Lysogeny versus lytic infection.
 
Our data suggest that a substantial proportion of bacteria in Lake Pavin contain functional viral genomes. Mitomycin C certainly does not induce all lysogenic bacteria and may also influence nonlysogens, although the concentration we used is known to induce temperate lambda phage while having only a small direct effect on its host bacterium, Escherichia coli (14). It is thus possible that our frequencies of lysogenic bacteria (range, 0.1 to 16%) are underestimates. Using the same methodological approach, lower values (0.1 to 7.4%) were reported for the surface layer of Lake Superior (17). Values of up to 84% have been reported for deep marine waters but generally are on average below 15% in estuarine, coastal, and offshore surface waters (21). In contrast to other viral and bacterial variables under study, lysogen levels were about fourfold higher in the oxic and relatively well-illuminated mixolimnion waters than in suboxic and anoxic waters (Table 1; Fig. 3). This indicates that the level of lysogeny, which was significantly related to depth, apparently was not affected by the surface light environment as expected from current knowledge. Contrary to their original hypothesis, Tapper and Hicks (17) also observed higher percentages of lysogenically infected bacteria in the surface water than in the subsurface water of Lake Superior. In our study, the relative abundances of lysogens were negatively correlated with bacterial abundance and production and with FIC and viral lytic production (Table 2), supporting the hypothesis that lysogeny is a strategy for survival of phages in environments with low host availability. This was also recently supported by a deep profiling study in the Mediterranean Sea, but with inversed vertical patterns compared to our study, i.e., increasing lysogens with decreasing FIC and the host abundance and productivity relative to depth (21). The relationship between the frequencies of lysogenically (y) and of lytically (x) infected bacteria in this study was described best by a power function [y = –4.5ln(x) + 20; r2 = 0.65], indicating that there are environmental characteristics, in relation to potential host densities and availability, favoring one of the two "viral life cycles" (21). The contrasting vertical patterns observed in this study and that of Weinbauer and coworkers (21) imply that the resource availability for phages (i.e., host environment) may be more important than the physical (i.e., temperature and light) or the chemical (i.e., O2) environments in favoring one of the two "viral life cycles."


arrow
ACKNOWLEDGMENTS
 
Jonathan Colombet was supported by a doctoral fellowship from the Grand-Duché de Luxembourg (Ministry of Culture, High School, and Research). The study was partly supported by the French National Program ACI/FNS "ECCO" (VIRULAC research grant awarded to T. Sime-Ngando).

We thank A. C. Lehours for technical and field assistance, J. Dolan for proofreading of an earlier version of the manuscript, and anonymous reviewers for helpful comments.


arrow
FOOTNOTES
 
* Corresponding author. Mailing address: Laboratoire de Biologie des Protistes, Université Blaise Pascal (Clermont-Ferrand II), UMR CNRS 6023, F-63177 Aubière Cedex, France. Phone: 33 4 73 40 78 36. Fax: 33 4 73 40 76 70. E-mail: Telesphore.SIME-NGANDO{at}univ-bpclermont.fr. Back


arrow
REFERENCES
 
    1
  1. Amblard, C., and G. Bourdier. 1990. The spring bloom of the diatom Melosira italica subsp. subarctica in Lake Pavin: biochemical, energetic and metabolic aspects during sedimentation. J. Plankton Res. 12:645-660.[Abstract/Free Full Text]
    2. 2
  2. Bettarel, Y., T. Sime-Ngando, C. Amblard, and J. Dolan. 2004. Viral activity in two contrasting lake ecosystems. Appl. Environ. Microbiol. 70:2941-2951.[Abstract/Free Full Text]
    3. 3
  3. Bettarel, Y., T. Sime-Ngando, C. Amblard, J.-F. Carrias, and C. Portelli. 2003. Virioplankton and microbial communities in aquatic systems: a seasonal study in two lakes of differing trophy. Freshwater Biol. 48:810-822.[CrossRef]
    4. 4
  4. Carrias, J.-F., A. Thouvenot, C. Amblard, and T. Sime-Ngando. 2001. Dynamics and growth estimates of planktonic protists during early spring in Lake Pavin, France. Aquat. Microb. Ecol. 24:163-174.
    5. 5
  5. Carrias, J.-F., C. Amblard, and G. Bourdier. 1996. Protistan bacterivory in a oligomesotrophic lake: importance of attached ciliates and flagellates. Microb. Ecol. 31:249-268.[Medline]
    6. 6
  6. Hara, S., I. Koike, K. Terauchi, H. Kamiya, and E. Tanoue. 1996. Abundance of viruses in deep oceanic waters. Mar. Ecol. Prog. Ser. 145:269-277.
    7. 7
  7. Humayoun, B. S., N. Bano, and T. Hollibaugh. 2003. Depth distribution of microbial diversity in Mono Lake, a meromictic soda lake in California. Appl. Environ. Microbiol. 69:1030-1042.[Abstract/Free Full Text]
    8. 8
  8. Jiang, S. C., and J. H. Paul. 1996. Occurrence of lysogenic bacteria in marine microbial communities as determined by prophage induction. Mar. Ecol. Prog. Ser. 142:27-38.
    9. 9
  9. Kirschner, A. K. T., and B. Velimirov. 1999. Modification of the 3H-leucine centrifugation method for determining bacterial protein synthesis in freshwater samples. Aquat. Microb. Ecol. 17:201-206.
    10. 10
  10. Noble, R. T., and J. A. Fuhrman. 1998. Use of SYBR Green I for rapid epifluorescence counts of marine viruses and bacteria. Aquat. Microb. Ecol. 14:113-118.
    11. 11
  11. Ortmann, A. C., J. E. Lawrence, and C. A. Suttle. 2002. Lysogeny and lytic viral production during a bloom of the cyanobacterium Synechococcus spp. Microb. Ecol. 43:225-231.[CrossRef][Medline]
    12. 12
  12. Pedros-Alio, C., J. I. Calderon-Paz, and J. M. Gasol. 2000. Comparative analysis shows bacterivory, not viral lysis, controls the abundance of heterotrophic prokaryotic plankton. FEMS Microb. Ecol. 32:157-165.[Medline]
    13. 13
  13. Pradeep Ram, A. S., D. Boucher, T. Sime-Ngando, D. Debroas, and J.-C. Romagoux. 2005. Phage bacteriolysis, protistan bacterivory potential, and bacterial production in a freshwater reservoir: coupling with temperature. Microb. Ecol. 50:64-72.[CrossRef][Medline]
    14. 14
  14. Roberts, J., and C. Roberts. 1975. Proteolytic cleavage of bacteriophage lampda repressor in induction. Proc. Natl. Acad. Sci. USA 72:147-151.[Abstract/Free Full Text]
    15. 15
  15. Sanders, R. W., K. G. Porter, S. J. Bennett, and A. E. Debiase. 1989. Seasonal patterns of bacterivory by flagellates, ciliates, rotifers, and cladocerans in a freshwater planktonic community. Limnol. Oceanogr. 34:673-687.
    16. 16
  16. SCOR-UNESCO. 1966. Determination of photosynthetic pigments in sea water. Unesco, Paris, France.
    17. 17
  17. Tapper, M. A., and R. E. Hicks. 1998. Temperate viruses and lysogeny in Lake Superior bacterioplankton. Limnol. Oceanogr. 43:95-105.
    18. 18
  18. Weinbauer, M. G., and F. Rassoulzadegan. 2004. Are viruses driving prokaryotic diversification and diversity? Environ. Microbiol. 6:1-11.[CrossRef][Medline]
    19. 19
  19. Weinbauer, M. G. 2004. Ecology of prokaryotic viruses. FEMS Microbiol. Rev. 28:127-181.[CrossRef][Medline]
    20. 20
  20. Weinbauer, M. G., and M. G. Höfle. 1998. Significance of viral lysis and flagellate grazing as factors controlling bacterioplankton production in a eutrophic lake. Appl. Environ. Microbiol. 64:431-438.[Abstract/Free Full Text]
    21. 21
  21. Weinbauer, M. G., I. Brettar, and M. G. Höfle. 2003. Lysogeny and virus-induced mortality of bacterioplankton in surface, deep, and anoxic marine waters. Limnol. Oceanogr. 48:169-177.
    22. 22
  22. Wetzel, R. G., and G. E. Likens. 1995. Limnological analysis, 2nd ed. Springer-Verlag, New York, N.Y.
    23. 23
  23. Wilhelm, S. W., and C. A. Suttle. 1999. Viruses and nutrient cycles in the sea. BioScience 49:781-788.[CrossRef]
    24. 24
  24. Wommack, K. E., and R. R. Colwell. 2000. Virioplankton: viruses in aquatic ecosystems. Microbiol. Mol. Biol. Rev. 64:69-114.[Abstract/Free Full Text]

Applied and Environmental Microbiology, June 2006, p. 4440-4445, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.00021-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.




This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Colombet, J.
Right arrow Articles by Demeure, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Colombet, J.
Right arrow Articles by Demeure, G.
Agricola
Right arrow Articles by Colombet, J.
Right arrow Articles by Demeure, G.

Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»