3042/03/89

Journal of Applied Bacteriology 1990,68, 189-198

Survival strategy of Escherichia coli and Enterococcus faecalis

in illuminated fresh and marine systems
I . B A R C I N A *J,. M . GONZALEZ,
J . I R I B E R R&
I L. EGEADepartmento de
Microbiologia e lnmunologia, Facultad de Ciencias, Universidad del Pais Vasco, Apdo 644,
48080-Bilba0, Spain
Accepted 8 June 1989
B A R C I N A ,I., GONZALEZ,
J.M., I R I B E R R I , J. & EGEA,L. 1990. Survival
strategy of Escherichia coli and Enterococcus faecalis in illuminated fresh and
marine systems. Journal of Applied Bacteriology 68, 189-198.
Some effects of visible light on Escherichia coli and Enterococcus faecalis in natural
freshwater and seawater were studied by plate counts, colony area measurements,
and direct counts. A large number of somnicells (non-culturable cells) were noted in
illuminated systems as compared with non-illuminated ones. Colony areas were significantly smaller in illuminated systems. Indirect activity measurements were used
to test the effects of visible light on the ability of E. coli and Ent. faecalis to
metabolize substrates ([14C]glucose) in natural waters. In illuminated systems, a
decrease of glucose uptake was observed. When percentages of assimilation and
respiration with respect to the total glucose uptake were analysed a decrease of
assimilation percentages and an increase of respiration percentages were observed.
In addition, differences in glucose uptake, assimilation and respiration by enteric
bacteria were detected for E. coli at the beginning of the experiments between freshand seawater and these were interpreted as a toxic effect exerted by seawater on E.
coli cells. Differences between species, natural waters and parameters studied
(excepting glucose assimilation) were detected in the illuminated systems. We concluded, however, that enteric bacteria under visible light illumination show a
general survival strategy characterized by reaching progressively a somnicell stage
which can be defined in terms of their (1) inability to form colonies on standard
bacteriological media, (2) inability to incorporate substrates, and (3) inactivation of
biosynthetic processes

Enteric bacteria in natural aquatic ecosystems

are affected by different factors. Bacterial competition (Jannasch 1968), predation (Enzinger &
Cooper 1976; McCambridge & McMeekin
1979, 1981; Barcina et al. 1986a), temperature
(McFeters & Stuart 1972; Davenport et al.
1976; Anderson et al. 1983; Gameson 1984;
Barcina et al. 1986a), nutrient concentration
(Hendricks & Morrison 1967; Hendricks 1972;
Barcina et al. 1986b), light and other physical
and chemical parameters have been cited as
responsible factors for decrease in colony counts
in aquatic ecosystems.
Physical and chemical parameters have been
cited by some authors (Sieburth & Pratt 1962;

* Corresponding author.

Verstraete & Voets 1975; Rhodes et al. 1983;

Munro et al. 1987; Gauthier et al. 1987) and
their effects have been studied mainly by monitoring the evolution of enteric bacterial colony
counts during an incubation period of several
days. Other methods, however, such as indirect
activity measurements, are not usually used on
enteric bacteria in natural aquatic media.
Visible light has been mentioned as an important factor affecting enteric bacteria survival in
natural waters (Gameson & Saxon 1967; Jagger
1975; Krinsky 1976; Grigsby & Calkins 1979;
Kapuscinski & Mitchell 1981; McCambridge &
McMeekin 1981; Fujioka et al. 1981; Barcina et
al. 1986b). Until now, most of the experiments
have compared only the changes in numbers of
colony-forming units (cfu) of enteric bacteria.

I . Barcina et al.
Recently, however, direct count techniques
such as acridine orange direct count (AODC
method) (Hobbie et al. 1977), have been shown
that cfu counts do not give good estimates of
the numbers of enteric bacteria present in
natural aquatic systems where different stressful
factors can induce an inability to form colonies
on standard bacteriological media (Xu et al.
1982; Grimes et al. 1986; Roszak & Colwell
1987; Lopez-Torres et al. 1988). Visible light is
one of those factors (see above) and there may,
therefore, be an important fraction of nonculturable enteric bacteria present in those
natural aquatic systems.
Roszak & Colwell (1987) studied the metabolic effect of a natural aquatic system on
Escherichia coli and Salmonella enteritidis and
defined the viviform population as the whole of
bacteria counted by a direct count method (the
AODC method). In that viviform population
they distinguished two major groups of cells:
culturable cells and somnicells. The former are
formed by cells which were able to form colonies on standard bacteriological media and the
somnicells are non-culturable cells. Barcina et
al. (1989a) studied E . coli survival under visible
light illumination in freshwater and showed that
after 72 h most of E . coli cells were in a somnicell stage; that is, they were non-culturable cells.
Activity measurements have been little used
to show physiological behaviour of enteric bacteria in illuminated natural aquatic media by
visible light. Indirect activity measurements
were proposed by Barcina et al. (1989b) to test
the effect of visible light on E. coli cells in freshwater, but we know of no studies that compared
these techniques with the effects of visible light
on different species of enteric bacteria in different natural aquatic systems.
The aim of this work was to study the effects
of visible light on enteric bacteria in natural
aquatic systems in respect of their ability to
form colonies on suitable culture media, and
their ability to metabolize glucose in these
environmental conditions. Indirect activity measurements were also used to check the effect of
freshwater and seawater on enteric bacteria.
Materials and Methods
This study was carried out with natural water
samples from the Butron river (Spain) and La
Salvaje beach, 500m offshore (Spain). All
samples were collected from the surface.

MICRO-ORGANISMS

Bacterial strains employed in this study were

Escherichia coli ATCC 11775 and Enterococcus
faecalis ATCC 19433.
INOCULA PREPARATION

The organisms were grown in nutrient broth at

28C for 8 h. Cells from the exponential phase
were harvested by centrifugation (3000 g for 15
min) and washed three times with sterile saline
solution (0.9% w/v). The pellets were suspended
in saline solution, inoculated in fresh nutrient
broth, and incubated at 28C for 18 h. The cells
were harvested at stationary phase as described
above. The final suspensions were inoculated in
the freshwater or seawater samples to give a
final density of approximately 10' cells/ml.

T E C H N I C A L METHODS

All experiments were carried out in 2 1 flasks

containing 500 ml of sterile subsamples,
obtained by filtering natural water through
membrane filters (0.2 pm; Millipore). The incubation of inoculated subsamples was done at
20C with shaking at 180 rev/min in an orbital
incubator with an illumination system consisting of seven Sylvania F30 W/T8/D lamps. The
luminous spectrum of these lamps is shown in
Fig. 1. The following parameters were studied
both in illuminated and in non-illuminated subsamples.

10

~6

4
2
0

350

450

550

650

750

nm

Fig. 1. Luminous spectrum of the lamps employed.

Survival strategy of enterobacteria

PLATE C O U N T S

Colony forming units (cfu) of E. coli and Ent.

faecalis were enumerated on Trypticase Soy
Agar (BBL) supplemented with 0.3% yeast
extract and 0.5% glucose (TSY) (Rhodes et al.
1983) incubated at 37C for 24 h. The areas of
the colonies of E. coli and Ent. faecalis on TSY
after incubation at 37C for 24 h were calculated by measuring colony diameters and
assuming that the colonies were all circular.
ACRIDINE ORANGE DIRECT COUNTS

The total number of cells in the subsamples was

estimated by the AODC method of Hagstrom et
al. (1979). Subsamples were preserved in 2%
formaldehyde (final concentration).
I N D I R E C T A C T I V I T Y MEASUREMENTS

Indirect activity measurements were done with

[U-'4C]glucose (250 mCi/mmol; Radiochemical
Centre, Amersham, England) at a saturated concentration (100 pgC/l). Three 5 ml subsamples
were analysed for each uptake measurement,
and they were incubated for 30 min in the dark
at 20C with shaking. Forty p1 of 2 N H2S04
were then injected into the flasks and the subsamples were further incubated for 1 h to trap
the 14C0, released on a filter paper impregnated with 1-phenetilamine (Hobbie & Crawford 1969). After both incubation periods, the
entire volume of each subsample was filtered
through membrane filters (0.2 pm; Millipore).
The filters were rinsed three times with 5 ml of
filtered water (0.2 pm membrane filters), placed
in Unisolve-1 (Hispanoland, Barcelona, Spain),
and radioassayed by liquid scintillation
counting. Quench curves were computed by the
channel ratio method. Controls for abiotic
absorption were prepared.
Total ['4C]glucose uptake was defined as
the sum of assimilation and respiration of
[14C]gIucose. Percentage of total [14C]glucose
uptake with respect to the [14C]glucose added
was calculated by the following expression :
Total ['4C]glucose uptake
x 100
[14C]glucose added
Assimilated and respired fractions were
defined as Wright & Burnison (1979). Respiration (or mineralization) is defined as the pro-

191

duction of 14C0, and assimilation as the

substrate retained in the cell (that is, substrate
taken up but not respired). Percentages of
assimilation and respiration with respect to
total [14C]glucose uptake were obtained by
dividing the assimilated or respired fraction by
the total ['4C]glucose uptake and then multiplying by 100.
STATISTICAL ANALYSIS

Colony areas were compared by Student's t-test

according with Sokal & Rohlf (1969). Regression analyses between the different parameters
and incubation times were used throughout this
study, and slopes, if different from zero, were
compared to see if there were any differences
between the effect of visible light on different
parameters, natural media and enteric bacteria
species. Any probability (P) less than 0.05 was
considered significant.
Results
The following results correspond to means of
three experiments carried out for E. coli and
Ent. faecalis in both natural aquatic media.
D I R E C T A N D CfU C O U N T S

In all cases both direct and cfu counts carried

out in non-illuminated systems did not show
any significant changes during the experiments.
In illuminated systems, however, cfu counts of
both E. coli and Ent. faecalis decreased during
the incubation period although total number of
bacteria (AODC method) remained constant
during the experiments done in illuminated
systems. Culturable and somnicell fractions of
viviform population for E . coli and Ent. faecalis
in both aquatic media for illuminated and nonilluminated systems are compared in Fig. 2.
There was a decrease in the ratios of cfu of E .
coli to cfu of Ent.faecalis throughout the experiments in illuminated systems (Table 1). For seawater, this was by a factor of 2 at 24 h and 7 at
48 h, and for freshwater, by a factor of 30 and
229, for 24 and 48 h respectively. However, in
non-illuminated systems very small differences
were observed (Table 1).
During these experiments, colony areas of cfu
on TSY were measured and there was a progressive diminution in illuminated as opposed
to non-illuminated systems for both E . coli and

192

I . Barcina et al.

Fig. 2. Fractions of somnicells ( 0 )and culturable cells (m) from the viviform population of Escherichia coli and
Enterococcusfaecalis in non-illuminated (a) and illuminated (b) systems. A. Escherichia coli in seawater; B. Enterococcusfaecalis in seawater; C . Escherichia coli in freshwater; D. Enterococcusfaecalis in freshwater.

Ent. faecalis in fresh- and seawater. These

results are shown in Table 2 as percentages of
illuminated to non-illuminated systems and
indicate a decrease from ca 100% to about
32-34% for E. coli and 4&50% for Ent.faecalis.
There were also differences between the colony

areas of illuminated and non-illuminated

systems. Thus there were significant differences
at 12 h for E . coli (P < 0.001) and Ent. faecalis
(P < 0.05) in freshwater, and at 24 h in seawater
(P < 0.001 and P < 0.01 for E . coli and Ent. faecalis respectively) (Table 2).

Table 1. Ratios of Escherichia coli to Enterococcusfaecalis during illuminated and

non-illuminated experiments in fresh and marine systems

In seawater

In freshwater

Time
(h)

Illuminated

Non-illuminated

Illuminated

Non-illuminated

0
12
24
36
48

5.11
4.03
2.82
1.46
0.70

3.52
2.68
2.54
2.90
1.70

3.32
1.1 1
0.19
0.01
0.01

3.52
6.92
3.91
2.70
3.19

Survival strategy of enterobacteria

193

Table 2. Percentages of colony areas for illuminated cells with respect to non-illuminated cells during the experiments carried out in fresh- and seawater
In freshwater

In seawater
Escherichia coli

Enterococcus fuecalis

Escherichia coli

Enterococcus faecalis

Times
(h)

Percentage

P*

Percentage

P*

Percentage

P*

Percentage

0
12
24
36
48

100.52
87.77
58.16

NS
NS

95.01
79.03
68.21

NS
0.001

0.001

NS
NS
0.01

101.51
96.27
85.90
67.63
39.99

92.39
94.07
92.47
54.22
50.31

34.30

0.001

0.001

0.001

0.0 1

32.36

0.00 1

P*

NS
0.05
0.0 1
0.001
0~001

* Significance level.
INDIRECT ACTIVITY MEASUREMENTS

Table 3 shows total uptake of ['4C]glucose for

illuminated and non-illuminated systems. For
illuminated cells, there was a sharp decrease in
the total uptake of glucose. In non-illuminated
cells the decreases were smaller or there was no
change.
It is important to note that E . coli cells in
freshwater took up much more [14C]glucose
(about 2.4 x lo6 cpm) than in seawater (about
lo5 cpm) at the beginning of the experiments.
However, Ent. faecalis took up similar quantities of [ 14C]glucose in both natural aquatic
media (about 5-6 x lo5 cpm). Percentages of
glucose uptake are given in Table 3.
The results of assimilation and respiration
percentages with respect to the total glucose
uptake are shown in Table 4. This shows
decreases of assimilation percentages and
increases of respiration percentages in all illuminated systems studied which were not observed
in the non-illuminated systems.
There were differences in the glucose assimilation and respiration percentages of E . coli at
the beginning of the experiments carried out in
freshwater but not in seawater (Table 4). For E .
coli cells in freshwater the percentages of
glucose assimilation and respiration were about
86% and 14%, respectively at 0 h and E . coli in
seawater presented percentages about 24% and
76%, respectively, also at 0 h. On the other
hand, Ent. faecalis did not give such high differences as E . coli.

DIFFERENCES BETWEEN PARAMETERS,

SPECIES A N D N A T U R A L WATERS

In illuminated systems, it was noted tha: semilog representations of decreases of cfu numbers,

total uptake of glucose, and assimilated and

respired fractions of glucose taken up vs time
were linearly related ( P < 0.05). Slopes of these
were significantly different from zero ( P < 0.05)
and were used to compare decreases of the
parameters studied. Slopes for non-illuminated
systems and direct counts (AODC method) were
not significantly different from zero.
Significant differences between cfu number
decrease and the corresponding decreases of
total
uptake
( P < 0.001),
assimilation
( P < 0.025) and respiration ( P < 0.001) of
glucose were observed in all cases except for
Ent. faecalis in illuminated marine systems; in
these we observed only significant differences
between cfu numbers and assimilation of
glucose decreases ( P < 0.05).
Comparison of slopes of faecal indicator bacteria in different natural aquatic media showed
that the decrease of E . coli cfu numbers was significantly lower ( P < 0.001) in illuminated-fresh
than in illuminated-marine waters. For Ent. faecalis in illuminated systems, decrease of cfu
numbers was not significantly different in freshand seawater but, in this case, it was seen that
slopes of total uptake ( P < 0.001) and respiration ( P < 0.001) of glucose were significantly
lower in sea- than in freshwaters. No differences
were found, either for E. coli or for Ent. faecalis,
with respect to glucose assimilation decrease.
When slopes between E . coli and Ent. faecalis
in a single natural aquatic medium were compared, there were significant differences in cfu
number decrease in illuminated-seawater
( P < 0.001) and in illuminated-freshwater
( P < 0905) systems, descending faster with E .
coli than with Ent. faecalis. Respiration of
glucose decrease was significantly different
( P < 0.001) between E . coli and Ent. faecalis in
both illuminated systems and decrease of total

71.88
71.09
67.12
64.38
61.32

28.12
28.91
32.88
35.62
38.68

0
12
24
36
48

Nonilluminated

23.61
14.33
12.95
5.00
2.25

76.39
85.67
87.05
95.00
97.75

R
41.82
50.84
45.97
46.08
54.84

58.18
49.16
54.03
53.92
45.16

R
40.67
52.92
51-10
71.63
83.05

A
59.33
47.08
48.90
28.37
16.95

Illuminated

Enterococcus faecalis
Nonilluminated

In seawater

Illuminated

Escherichia coli

(h)

Time

13.61
7.13
4.78
2.27
1.68
56.10

56.57
58.26
63.69
65.12

Nonilluminated
57.66
34.75
19.62
5.61
5.75

Illuminated

Escherichia coli

11.56

11.24
8.16
8.27
9.32

R
12.93
11.28
13.43
19.31
25.11

A
87.07
88.72
86.57
80.69
74.89

Nonilluminated

85.02
49.06
39.61
12.32
9.92

14.98
50.94
60.39
87.68
90.08

Illuminated

Eschmichia coli

37.91
18.00
19.80
26.89
44.42

55.58

62.09
82-00
80.20
73.11

404M
7.34
7.07
19.98
14.07

92.66
92.93
80.02
85.93

60.00

Illuminated

Enterococcus faecalis
Nonilluminated

In freshwater

14.26
5.03
2.63
0.70
0.42

Illuminated

Enterococcus faecalis

Nonilluminated

In freshwater

Table 4. Percentages of assimilation (A) and respiration (R)with respect to total ['4C]glucose uptake in illuminated and non-illuminated systems

12.81
9.73
9.21
7.54
8.51

Illuminated

Enterococcus faecalis

Nonilluminated

Total [14C]glucose uptaken

x 100.
['4C)glucose added

2.21
1.68
1.86
0.64
0.30

2.72
1.65
1.99
2.08
2.05

0
12
24
36
48

* Total uptake =

Illuminated

Nonilluminated

Time
(h)

Escherichia coli

In seawater

Table 3. Percentage of total r'4C1glucose uptake during experiments in fresh and marine water*

11

\o

Survival strategy of enterobacteria

glucose uptake was in fresh- ( P < 0.025) and in
seawater ( P < 04)05) illuminated systems.
However, respiration and total uptake of
glucose of E . coli decreased faster than Ent. faecalis in illuminated seawater but this was
opposite in illuminated freshwater. No differences for glucose assimilation were found
between E. coli and Ent. faecalis in illuminated
fresh- or seawater.

Discussion
D I R E C T A N D CfU C O U N T S

Differences between direct (AODC method) and

cfu counts provide information about the fractions of somnicells and culturable cells which
constitute the viviform populations in our
aquatic systems. Figure 2 shows these fractions
and it can be seen that visible light induces a
progressive dormancy of both E. coli and Ent.
faecalis in fresh- and in seawater, detected
because most of the cells lost their ability to
form colonies on a suitable culture medium.
Such organisms are called somnicells. In illuminated systems, fractions of culturable cells from
viviform populations are very small and standard bacteriological media should not therefore
be used to detect numbers of enteric bacteria in
natural waters exposed to visible light because
this procedure would give an important underestimation of enteric bacteria real numbers.
Significant differences ( P < 0.001) in colony
areas (Table 2) have been found throughout the
experimental period between illuminated and
non-illuminated systems. This would represent a
stress effect of visible light on both E . coli and
Ent. faecalis in natural waters and could be a
first step towards the loss of ability to form
colonies on standard bacteriological media, i.e.
towards the somnicell stage.
It is concluded, therefore, that visible light has
a stressful effect on enteric bacteria in natural
aquatic media. However, Fujioka e t al. (1981)
indicated, for seawater, that visible spectrum of
sunlight 'has a killing rather than a stressful
effect on the indicator bacteria' (faecal coliforms
and faecal streptococci). Other authors
(Hollaender 1943; Gameson & Gould 1974;
Krinsky 1976) used the terms lethal and death
when referring to decrease in cfu number.
Nevertheless, these authors did not make direct
counts to determine total number of enteric bac-

195

teria (viviform population) in their experiments.

Therefore, the terms killing, lethal and death
have been interpreted by them as a decrease in
cfu numbers and they did not consider them to
be a decrease in total cell numbers of a viviform
population. At present, according with several
authors (Grimes et al. 1986; Roszak & Colwell
1987; Barcina e t al. 1989) these terms (killing,
lethal and death) are understood as a decrease
in bacterial total number. Caution is therefore
necessary in interpreting existing literature, particularly that published before eighties.
A comparison of the decrease of cfu numbers
of E. coli and Ent. faecalis during our experiments shows a more rapid inactivation of E . coli
as opposed to Ent. faecalis both in marine and
in freshwater exposed to visible light. This
results in a rapid reduction of the E . coli to Ent.
faecalis cfu ratio (Table 1) in illuminated
systems although no important decline in the
ratio of E . coli to Ent. faecalis in nonilluminated systems was observed. These results
indicate that the significance of the faecal coliforms to faecal streptococci ratio established by
Geldreich & Kenner (1969) for 24 h after input
of faecal indicator bacteria in natural waters is
not valid in natural aquatic ecosystems where
visible light can affect enteric bacteria survival.
I N I T I A L EFFECT O F S E A W A T E R
VERSUS F R E S H W A T E R

Several authors (Zobell 1936; Carlucci &

Pramer 1959; Vasconcelos & Swartz 1976;
Rhodes et al. 1983; Munro et al. 1987) have
observed a toxic effect of seawater on enteric
bacteria. They showed, that in the absence of
natural microflora, there was a decrease in
numbers of enteric bacteria in seawater during
their incubation periods.
In our experiments we noted that E . coli in
freshwater took up much more ['4C]glucose
than in seawater (Table 3). However, Ent. faecalis showed similar incorporation
of
['4C]glucose in freshwater and in seawater
(Table 3). That means that seawater has a
greater effect on transport of glucose by E. coli
cells than does freshwater. Moreover, percentages of assimilation of E . coli in freshwater at
0 h were very high compared with those in seawater (Table 4).However, for Ent. faecalis differences in
assimilation and
respiration
percentages were not as important as for E. coli

196

I . Barcina et al.

(Table 4). By that, we can deduce E. coli is more

sensitive to natural waters than is Ent. faecalis,
and that natural waters can affect enteric bacteria metabolism. Moreover, these effects were
produced immediately after input of enteric bacteria in natural waters. In spite of these results,
cfu numbers of both E . coli and Ent. faecalis
were not affected by natural fresh- and seawater
at the beginning of the experiments because the
density of these enteric bacteria were similar.
By these facts we can conclude that seawater
has several effects on E. coli cells which can be
shown metabolically and immediately (without
long incubation periods) by using indirect activity measurements, and not only by counts
throughout a long incubation period (usually
several days) as other authors (Zobell 1936;
Carlucci & Pramer 1959; Rhodes et al. 1983;
Munro el al. 1987) have shown. For E . coli in
seawater with respect to freshwater, these effects
are a decrease of total glucose uptake and a use
of that glucose taken up mainly by degradative
processes (respiration). This could be interpreted
as a mechanism of resistance of E . coli to seawater.

I N D I R E C T ACTIVITY MEASUREMENTS
UNDER ILLUMINATION

Progressive decrease of total glucose uptake by

illuminated cells (Table 3), both in fresh- and
seawater, indicates an inactivation of glucose
transport into the cells as a consequence of
visible light action.
If we analyse percentages of assimilation and
respiration with respect to total glucose uptake
(Table 4) we can observe a progressive inactivation of biosynthetic process throughout the
illuminated experiments for both E . coli and
Ent. faecalis in fresh- and seawater. This means
that the glucose taken up is mainly used in
degradative process, that is, enteric bacteria in
illuminated natural waters become dormant
cells which are characterized by using substrates
taken up for maintenance functions and by
inactivating their biosynthetic processes.
Roszak & Colwell (1987) studied evolution of
E . coli and Salmonella enteritidis in marine dark
systems and observed throughout their incubation period (1 or 2 months) an increase in the
number of cells which incorporated substrates
for maintenance functions but not for growth.

From these results we see that visible light lead

to progressive dormancy of enteric bacteria
more rapidly than occurs in natural aquatic
media in the dark, and therefore we can aftirm
that the effects of visible light on enteric bacteria
are additional to those of natural aquatic media.
From the results so far discussed, we can
deduce that visible light stimulates in E . coli and
Ent. faecalis cells in natural aquatic media different metabolic effects which lead to a progressive dormancy. This progressive dormancy lead
enteric bacteria to a somnicell stage which can
be defined and characterized by (1) inability to
form colonies on standard bacteriological
media, (2) incapacity to incorporate substrates,
and (3) inactivation of biosynthetic processes
and a metabolism of maintenance.

DIFFERENCES BETWEEN PARAMETERS,

SPECIES A N D N A T U R A L W A T E R S

We have found differences between enteric bacterium species and between each species in the
two ecosystems. This means that visible light
effects depend on the natural aquatic ecosystem
studied and on enteric bacteria species used.
Differences in the effect of visible light on the
decrease in cfu for different enteric bacteria and
natural aquatic media have already been noted
by Fujioka et al. (1981). In our study, we point
out differences both for the decrease in cfu and
for indirect activity measurements (respiration
and total uptake of glucose). With respect to
these indirect activity measurements it is important to note that no differences have been
obtained for glucose assimilation decrease
between E . coli and/or Ent. faecalis in fresh
and/or seawater. By this fact, we can deduce
that the effect of visible light on assimilative
processes of enteric bacteria in natural waters
could be independent of the enteric bacteria
species and the natural aquatic media studied.
These facts suggest that visible light affects
the parameters studied with different intensity in
each case (excepting glucose assimilation) and
bacteria of enteric origin may respond to illuminated natural aquatic systems in a speciesspecific manner. In spite of those differences,
enteric bacteria show a general survival strategy
characterized by reaching the somnicell stage as
defined above.

Survival strategy of enterobacteria

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