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The Significance of Viruses to Mortality in Aquatic Microbial

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Article in Microbial Ecology · September 1994

DOI: 10.1007/BF00166813 · Source: PubMed

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Microb Ecol (1994) 28:237-243
Controls of the Microbial Loop:
/ CROBIAL
Biotic Factors ECOLOGYInc.
© 1994Springer-Verlag
New York

The Significance of Viruses to Mortality in Aquatic

Microbial Communities

C.A. Suttle
Marine Science Institute, The University of Texas at Austin, P.O. Box 1267, Port Aransas, Texas
78373-1267, USA

Abstract. A variety of approaches including enumeration of visibly infected

microbes, removal of viral particles, decay of viral infectivity, and measure-
ments of viral production rates have been used to infer the impact of viruses on
microbial mortality. The results are surprisingly consistent and suggest that, on
average, about 20% of marine heterotrophic bacteria are infected by viruses
and 10-20% of the bacterial community is lysed daily by viruses. The effect of
viruses on phytoplankton is less certain, but ca. 3% of Synechococcus biomass
may be lysed daily. The fraction of primary productivity this represents de-
pends upon the relative biomass and growth rate of Synechococcus. Virus
enrichment experiments suggest that the productivity of eukaryotic phyto-
plankton would be ca. 2% higher in the absence of viruses. Overall, probably
about 2-3% of primary productivity is lost to viral lysis. There is considerable
variation about these estimates; however, they represent a starting point for
incorporating viral-mediated processes into aquatic ecosystem models.

Introduction

An ever-growing number of studies have attested to the abundance and ecological

significance of viruses in aquatic environments [5]; yet there are no methods that
provide direct measurements of in situ rates of viral-mediated mortality. Nonethe-
less, a number of independent approaches can be used to infer the proportion of
microbial mortality that can be attributed to viruses. Such estimates can be used to
incorporate viral processes into aquatic ecosystem models.

Estimation of the Impact of Viruses on Aquatic Microbial Communities

Electron Microscopy of Natural Microbial Communities
About 0.9-4.3% (mean, 3.2%) of marine bacteria contain mature virions, with the
lowest percentage observed in oligotrophic waters [10]. As mature viruses are
visible only in the final stages of the lytic cycle, the proportion of infected cells
would be about 3.7- to 7.1-fold greater [1 t]. Using the average percent of infected
238 C.A. Suttle

Table 1. Averagedecay rates of viruses in natural seawater samples (SD, standard deviation; n,
number of replicatedexperiments)
Virus Host Decay rate
strain Family habitat (day- 1) SD n Reference
PWH3a-P1 Myoviridae Marine 0.300 0.0268 4 16
LB1VL-Plb Podoviridae Marine 0.600 0.0480 4 16
T2 Myoviridae Enteric 0.353 0.0245 2 1
T2 Myoviridae Enteric 1.225 0.5366 8 6
T7 Podoviridae Enteric 0.488 0.2313 2 2
nt-6 Myoviridae Estuarine 0.097 1 19
nt- 1 Podoviridae Estuarine 0.197 1 19
H7/2 Myoviridae Marine 0.528 0.1800 7 8
H40/1 Styloviridae Marine 0.528 0.2640 8 8
H6/1 Podoviridae Marine 0.456 0.3048 3 8

cells (3.2%) and conversion factor (5.4) suggests that ca. 17% of bacteria are
infected by viruses. Reasoning that, on average 50% of the cells that are produced
will die [10], then something that removes 50% of the cells should, on average,
account for 100% of the mortality. Viruses, therefore, would account for about
34% of the total mortality of bacteria [10]. If production is balanced by removal,
34% of the mortality would also represent 34% of the production. This approach is
attractive because infected cells are quantified by direct count; however, conver-
sion factors that are subject to considerable uncertainty must be used to estimate the
proportion of infected cells and viral-mediated mortality.

Decay Rates of Viruses in Seawater

Another approach is to assume that because virus abundance remains relatively

constant, virus production can be estimated from the virus removal rate when virus
production has been stopped [3, 7]. As viruses are produced at the expense of host
cells, if the number of viruses produced per lytic event is known, one can infer
mortality. Yet, removal rates estimated from direct counts by transmission electron
microscopy (TEM) are extremely rapid and cannot be balanced by measured
bacterial production rates [3, 7], suggesting that this method cannot be used to
estimate in situ rates of virus production.
Alternatively, as viruses infecting a wide variety of marine and nonmarine
bacteria have similar decay rates in natural waters (Table 1), the decay of infectivity
of virus tracers can be used to infer turnover rates of natural virus communities
[16]. Decay rates of infectivity should be regarded as maximum rates for the decay
of virus particles, because infectivity can be destroyed without removing virus
particles. Virus-specific production rates in coastal waters inferred from the aver-
age of the decay rates of virus infectivity in Table 1 are about 0.48 d a y - i (_+0.292);
hence, ca. 4.8 × 106 viruses m1-1 day - I would be produced if there were 107
viruses m l - i. Assuming that 50 viruses are produced per lytic event [7], ca. 96,000
(4.8 x 106/50) bacteria would be lysed daily, or about 9.6% of the standing stock
(assuming l0 6 bacteria ml-1). For bacteria growing at 0.5 day - I , this implies that
Viruses and Microbial Mortality 239

ca. 20% of the bacterial production is removed by lysis and, as the latent period is
typically similar to the generation time of uninfected hosts [ 11], ca. 20% of bacteria
should be infected by viruses.

Direct Measurements of Virus Production Rates in Seawater

Recently, a radiotracer-based method was developed for determining virus produc-

tion rates [12, 13]. Although the method depends upon separating bacteria and
viruses by filtration, it provides an independent method for assessing virus produc-
tion. At two offshore stations with measurable production the rates were 0.03 and
0.011 day-1, while at nearshore and coastal stations with measurable production,
the average was 0.85 day 1 (SD 0.60). Assuming a burst size of 50, the percentage
of bacteria that would have to be lysed daily to support the measured viral produc-
tion rates ranged from 3.2 to 50% (mean, 21%) at the most offshore and nearshore
stations, respectively.

The Impact of Viruses on Primary Producers

Infectious viruses that cause lysis of phytoplankton are widespread and can be
extremely abundant [4, 15, 17, 18]. In marine Synechococcus, viruses are visible in
0.8-2.8% (mean, 1.5%) of cells [ 10]. If the viruses are visible for 50% of the lytic
cycle [18] then ca. 3% of Synechococcus are infected by viruses.
Alternatively, cyanophage production rates can be inferred from the decay rates
of virus tracers added to seawater. As the concentration of infective cyanophages is
assayed rather than the total number of virus particles [15, 18], it is reasonable to
assume that the decay rates reflect the production rate of infectious viruses. Along
an 83-kin transect that extended offshore from Port Aransas, Texas, the concentra-
tion of viruses in the mixed layer that infected Synechococcus (strain DC2) ranged
from 1.53 × 104 to 2.52 x 105 ml- 1 (Table 2). The virus production rates required
to balance estimated decay rates integrated over the mixed layer (range, 0.18-2.01
day -1) would lyse 0.3-6.5 % (mean, 3.7%) of Synechococcus cells on a daily basis.
This assumes that 250 viruses are produced per lyric event [15]; a smaller burst size
would result in a proportionally larger increase in the inferred virus production
rates.
As well, contact rates provide an upper limit to the number of cells that can be
infected [9]. Virus contact rates (R) = (Sh 2w d Dv) V P, where, Sh is the
Sherwood number for Synechococcus (1.01, dimensionless), d is cell diameter
(1.5 × 10 - 4 cm), D v is the diffusivity of the viruses (4.0 × 10 .8 cm 2 s-l), and P
and V are the concentrations of Synechococcus and infectious cyanophages, respec-
tively. Along the offshore transect 5-83% of Synechococcus would be contacted
daily by infectious cyanophages (Table 2). Clearly, contact rates are adequate to
support the inferred virus production rates. In contrast, cyanophage concentrations
in the northwest Atlantic appear to be lower and only about 4% of Synechococcus
would be contacted daily when virus concentrations were highest [18].
Virus-addition/photosynthetic-suppression experiments can also be used to ex-
amine the effect of viruses on phytoplankton. Material from the 2 to 200-rim size
240 C.A. Suttle

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Viruses and Microbial Mortality 241

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CONCENTRATION FACTOR ( x A m b i e n t )

Fig. l. The effect of concentrated material from the virus size fraction on photosynthetic rates of
natural phytoplankton communities. The y-intercept would be the photosynthetic rate in the absence of
the viral size fraction.

Table 3. Predicted rates of primary productivity in the absence of viruses based on experiments in
which a range in concentrations of natural virus communities was added to natural seawater samples.
Seawater was collected from the Marine Science Institute pier, Port Aransas, Texas

1° productivity Enhancement of
Date (~gC 1-1 h- l) y-intercept n re productivity (%)
14 Sep 93 23.6 (1.65) 1.0115 3 1.000 1.15
26 Sep 89 18.0 (2.30) 1.0127 4 0.968 1.27
15 Mar 90 80.6 (1.74) 1.0225 5 0.912 2.25
05 Apr 90 16.9 (0.47) 1.0370 3 1.000 3.70

fraction o f seawater can strongly depress photosynthesis in phytoplankton [14, 17].

The y-intercept o f a linear regression through the initial slope o f the suppression
curve represents the photosynthetic rate in the absence o f the virus size fraction
(Fig. 1). In four experiments the predicted rates o f primary productivity in the
absence of viruses were enhanced by an average o f 2.1% (range, 1.2-3.7%) (Ta-
ble 3), Autoradiography indicated that eukaryotes were affected the most [14],
possibly because photosynthetic rates in Synechococcus are not affected by viral
infection until shortly prior to cell lysis, or about 9 h post infection [ 15], longer than
in the photosynthetic-suppression experiments.

The Impact of Viruses on Photosynthetic and Heterotrophic

Microbial Communities

Studies based on thin sections of microbial communities, decay rates o f infectivity,

and radiotracer estimates o f viral production are surprisingly consistent and suggest
242 C.A. Suttle

that, on average, about 20% of marine heterotrophic bacteria are infected and
10-20% of bacteria are lysed by viruses on a daily basis. In oligotrophic waters the
percentages are likely less and in productive waters they may be higher, but a good
first approximation is that 15% of heterotrophic bacteria are lysed daily by viruses.
As virus production appears to be tied to bacterial biomass but not bacterial
production [13], the proportion of bacterial production that this represents will
depend strongly on growth rate.
The effect of viruses on primary producers is more difficult to ascertain. TEM
and decay rate data suggest that ca. 3% of Synechococcus is lysed daily. The
proportion of the productivity that this represents depends upon the contribution of
Synechococcus to phototrophic biomass and their relative growth rate. The best
evidence that primary production from eukaryotes is lost to viral lysis is from virus
addition experiments that suggest that primary productivity in the absence of
viruses would be elevated by about 2%. In addition, as Synechococcus doubles
about once per day, about 3% of the production from this group would be lost to
lysis. Overall, a reasonable estimate is that 2-3% of primary production is lost to
viral lysis.
The above is a first attempt to draw together data from a number of independent
approaches and define the effect of viruses on aquatic bacteria and phytoplankton.
Intuitively, however, the greatest impact of viruses on aquatic ecosystems is
through non-steady-state processes such as maintenance of species diversity and
through genetic exchange among microbial populations.

Acknowledgments. The author is grateful for the assistance of AM Chan and Dr. Garza in the
cyanophage studies. This research was supported by the Office of Naval Research (N00014-92-J-1676)
and the National Science Foundation (OCE-90188833). Contribution No. 910 of the Marine Science
Institute

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