European Journal of Phycology

ISSN: 0967-0262 (Print) 1469-4433 (Online) Journal homepage: https://www.tandfonline.com/loi/tejp20

Cryptoendolithic growth of the red alga Galdieria

sulphuraria in volcanic areas

Wolfgang Gross , Jan Küver , Gilbert Tischendorf , Nicolas Bouchaala &

Wilhelm Büsch

To cite this article: Wolfgang Gross , Jan Küver , Gilbert Tischendorf , Nicolas Bouchaala &
Wilhelm Büsch (1998) Cryptoendolithic growth of the red alga Galdieria�sulphuraria in volcanic
areas, European Journal of Phycology, 33:1, 25-31, DOI: 10.1080/09670269810001736503

To link to this article: https://doi.org/10.1080/09670269810001736503

Published online: 03 Jun 2010.

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 Eur. J. Phycol. (1998), 33 : 25–31. Printed in the United Kingdom 25

Cryptoendolithic growth of the red alga Galdieria sulphuraria

in volcanic areas

W O L F G A N G G R O S S1, J A N K U> V E R2, G I L B E R T T I S C H E N D O R F1, N I C O L A S B O U C H A A L A3

A N D W I L H E L M B U> S C H4

" Institut fuX r Pflanzenphysiologie und Mikrobiologie ; Freie UniversitaX t Berlin, KoX nigin-Luise-Strasse 12-16a, 14195 Berlin, Germany
# Max-Planck-Institut fuX r Marine Mikrobiologie, 28359 Bremen, Germany
$ Institut fuX r Experimentalphysik, Freie UniversitaX t Berlin, 14195 Berlin, Germany
% Institut fuX r Mineralogie ; Freie UniversitaX t Berlin, 14195 Berlin, Germany

(Received 30 January 1997 ; accepted 15 July 1997)

The habitat of the acido- and thermophilic red algae Galdieria sulphuraria and Cyanidium caldarium was examined in acidic hot sulphur
springs in the vicinity of Naples (Italy). These species grew on soil and rocks, but a large part of the populations was cryptoendolithic.
The endolithic algal layer (1–3 mm in thickness) was covered by amorphous silica (1–2 mm in thickness) containing traces of hydrotroilite
(FeS\nH O) and elemental sulphur. Organotrophic bacteria and fungi were not found in the algal layer. Light penetration measurements
#
showed that 0±1–1 % of the sunlight reached the upper part of the algal layer. Thus, low-light-adapted algae should be able to
perform some photosynthesis in this endolithic habitat. Under conditions where light is even more limited, e.g. in winter or after
darkening of the covering layer, many of the cells might not survive. Aqueous extracts of these algae are excellent growth substrates for
Galdieria sulphuraria. Therefore, we propose that compounds released from dead cells in the endolithic layer are used by the surrounding
Galdieria cells for heterotrophic metabolism. This would increase their chance of surviving prolonged periods under detrimental
conditions.

Key words : cryptoendolithic growth, Cyanidium caldarium, Galdieria sulphuraria, heterotrophy, sulphur springs.

Introduction colour on acidic soils in the vicinity of hot springs or in

their effluents (Smith & Brock, 1973). Belly et al. (1973)
Highly acidic environments formed by the oxidation of
have reported that these mats also contain bacteria
sulphide minerals or reduced sulphur compounds are
(Bacillus coagulans) and a fungus (Dactylaria galloparva).
known to be populated by a range of acidophilic and acid-
Sometimes these mats are covered by a 3–5 mm thick
tolerant bacteria. The best studied of these bacteria are the
layer of silica and quartz granules (Smith & Brock, 1973) or
thiobacilli (Pronk et al., 1990 ; Johnson et al., 1993). Most
a soft mineral deposit (Seckbach, 1994). While Cyanidium
of the bacteria are generally considered to be obligate
is an obligate autotroph, Galdieria can also grow mixo-
chemolithotrophs obtaining energy from the oxidation of
and heterotrophically on a variety of organic compounds
reduced sulphur compounds and ferrous iron. In addition
(De Luca et al., 1972 ; Gross & Schnarrenberger, 1995).
several heterotrophic bacteria that grow on organic
Heterotrophy is unusual for a red alga and may reflect a
compounds occur in the same habitat. These bacteria
special survival strategy of Galdieria. However, Smith &
might be important for the removal of organic compounds
Brock (1973) estimated that the concentration of organic
that have an inhibitory effect on autotrophic bacteria
material other than the algae themselves is too low to
(Harrison, 1984 ; Johnson et al., 1992).
support significant heterotrophy and that bacteria and
Few eukaryotic organisms can live under the extreme
fungi are most probably the main consumers of organic
conditions of a hot sulphur spring. At temperatures
matter. The studies by Brock and co-workers (Brock,
around 50 °C and pH values of less than 2±0, the
1978) at Yellowstone National Park (Montana) might
overwhelmingly dominant eukaryotes are two unicellular
concern an unusual situation. Therefore, we investigated
red algae of the family Cyanidiophyceae : Cyanidium
the natural habitat of Cyanidium and Galdieria in Italy with
caldarium (Tilden) Geitler and Galdieria sulphuraria (Gal-
special reference to heterotrophic growth conditions.
dieri) Merola (Merola et al., 1981). In some locations a
third species, Cyanidioschyzon merolae De Luca, Taddei &
Varano, has been reported to occur, but its contribution to Materials and methods
the total biomass appears to be small (De Luca et al., 1978).
Cyanidium and Galdieria form mats of a deep blue-green Sampling and sample treatment
Correspondence to : W. Gross. Samples were taken at the end of April 1995 at La
Published online 03 Jun 2010
W. Gross et al.
Solfatara and Pisciarelli (near Pozzuoli, Naples, Italy). For
the determination of autotrophic and heterotrophic bac-
teria, samples were taken from two sites at Pisciarelli. Site
1 is a small pool where Galdieria was present as indicated
by a blue-greenish layer on rocks and the sediment close
to the water surface. Site 2 was similar to site 1 except that
no algal growth was visible. From both sites a 1 : 2 mixture
of particulate material and water was placed in sterile 1 l
glass bottles, shaken vigorously, and used for the most
probable number (MPN)-dilution series.
Media and enumeration of autotrophic and heterotrophic
bacteria
The MPN technique was used to estimate the number of
autotrophic and heterotrophic bacteria at the two sam-
pling sites. The inoculation of the MPN tubes was
performed within 5 h of sampling. For enumeration of the
different bacteria and Galdieria, dilution series (1 : 10) were
carried out as triplicates in culture tubes containing 9 ml
medium, starting with 1 ml mixed sediment and water
from the original sample. The mineral medium contained
0±4 g KH PO , 0±4 g MgSO \7H O, 0±4 g (NH ) SO ,
# % % # %# %
20 mg CaCl , 1 ml of trace element solution (SL-10) and
# electron microscopy (SEM) with EDX and a polarizing
1 ml tungstate-selenite solution (Widdel & Bak, 1992) in a
total volume of 1 l. The pH was adjusted to 2±0 with
H SO . For growth of autotrophic bacteria, 50 mM
# %
FeSO , or a pea-sized amount of elemental sulphur or FeS
% #
was added from sterile stock solutions as the electron
donor. Heterotrophic growth was monitored by addition
of 5 mM galactose or 5 mM mannose as electron donor.
Medium without an electron donor served as a control.
Each set contained one additional uninoculated series as a
control to ascertain sterile working conditions. The tubes
were incubated at 50 °C for 6 weeks in the dark and
checked microscopically for growth using a Zeiss Axio-
phot microscope. MPN values were calculated for each set
of samples from statistical tables and the results were
expressed as cells per millilitre (American Public Health
Association, 1969).
Analysis of sugars and sugar alcohols by HPLC
Water samples were filtered (0±2 µm) immediately after
sampling. Soil and rock samples were placed in air-tight
containers, stored in a cold room and analysed within 1
week. Ions and organic compounds were extracted from
soil and rock samples by adding 500 ml of distilled water
to 100 g material. Samples were shaken vigorously for
10 min and then left for 1 h without shaking. The
supernatant was centrifuged for 30 min at 10 000 g and
the resulting supernatant filtered through glass wool
(Whatmann GF}D). The filtrate was used for the de-
termination of DOC (dissolved organic carbon), pH and
conductivity, as well as analysis of ions. An aliquot of
100 ml was de-ionized by passage through Dowex W-50
and AG 1 and the eluate concentrated to 2 ml in a rotary
evaporator. Concentrated samples were filtered (0±2 µm)
and an aliquot of 20 µl analysed by high-performance
26
liquid chromatography (HPLC). The content of sugars and
sugar alcohols in untreated as well as in de-ionized and
concentrated samples was analysed on an HPLC column
(BC-100, Ca#+, 7±8¬300 mm ; Benson, Reno, USA). The
eluant was double-distilled water and the column tem-
perature was 85 °C. The effluent of the column was
monitored at 50 °C by an RI detector connected to an
integrator unit. Compounds were identified by com-
parison with the retention time of standard sugars and
sugar alcohols.
Analysis of ions and DOC
The content of As, Pb, Cd and Hg was determined by
atomic absorption spectroscopy using standard methods.
Sulphate was determined by ion-exchange chromato-
graphy and DOC was estimated by converting the organic
material to CO .
#
Analysis of mineral composition of rocks
The determination of mineral composition of the sampled
rock types and layers (colour variation from light grey to
yellow or dirty dark brown) was made by scanning
microscope.
Electron microscopy
For SEM, pieces (about 1 cm#) of the upper rock layer with
the adhering algal cells on the subsurface side were fixed
for 2 h with glutaraldehyde (2±5 %) in sodium–potassium
phosphate buffer (100 mM, pH 6±8) including paraformal-
dehyde (2 %). After washing with phosphate buffer (0±1 M,
pH 7±0) the specimens were dehydrated with acetone.
Samples were incubated with Peldri II (50 %) in acetone
(v}v) at 28–30 °C (Zimmer, 1989), vacuum dried at
6¬10−& bar, and sputter-coated with gold–palladium.
SEM was performed with a Zeiss DSM 950 at 10 kV and
10 mm working distance.
Transmission measurements
The mineral layer above the algal mat was carefully
broken away by inserting a scalpel into the algal layer.
Pieces up to 2 cm# and about 2 mm thickness were
obtained. Algae were removed from the subsurface by
mild ultrasonic treatment. Transmission of light through
representative pieces of the upper layer was measured
using a xenon high-pressure lamp. Monochromatic light
in the wavelength range 250–800 nm was focused on the
sample in a spot about 2 mm in diameter. Transmitted
light was measured with a photo-diode. Readings were
corrected for the output of the xenon lamp.
Endolithic growth of Galdieria supplied with high silicate
concentrations
The upper part of a piece of sandstone taken from
Pisciarelli was inoculated with a few drops of a unialgal
Endolithic growth of Galdieria
suspension of Galdieria and placed in a tray partly filled
with diluted growth medium (1}10) supplemented with
1 % sodium metasilicate. The tray was covered with
Nescofilm except for a c. 10 cm# area of the surface of the
stone and illuminated with 80 µmol m−# s−" at 30 °C. A
similar set-up without the addition of sodium metasilicate
served as a control. Growth was monitored periodically
under a dissecting microscope.
Growth of Galdieria on acidic extracts from Galdieria
sulphuraria and Cyanidium caldarium
Cells were grown heterotrophically on 50 mM galactose
(Galdieria sulphuraria strain 002P and strain 074 : Pinto
et al., 1992) or autotrophically (Cyanidium caldarium strain
IPPAS P-509, Culture Collection of Microalgae, Inst. Plant
Physiol., Moscow) in 1 l Erlenmeyer flasks with 500 ml
medium on a rotary shaker (120 rpm) at 25 °C, either in
the dark or illuminated with white incandescent light at
80 µmol m−# s−". Cells were harvested in log-phase (auto-
trophic cultures) or in late stationary phase. They were
washed with distilled water and 2 ml of 1 N H SO per
# %
gram of packed cells was added. The sample was incubated
at 80 °C for 4 h and centrifuged for 10 min at 4000 g. The
supernatant was frozen at ®20 °C, thawed, and centri-
fuged again. The clear supernatant was passed through a
0±2 µm nylon filter and added to a mineral medium (Gross
& Schnarrenberger, 1995) under sterile conditions. This
medium was inoculated with 2 ml of autotrophic cells of
Galdieria sulphuraria strains 002P and 002S, isolated from
the sites at Pisciarelli and La Solfatara, respectively.
Cultures were maintained in the dark at 25 °C. Growth
was monitored by measuring light scattering at 800 nm.
Cell suspensions were diluted with water to give a reading
between 0±05 and 0±15.
Results
A comparison of the physical and chemical parameters of
five different locations at Pisciarelli (near Pozzuoli, Naples)
and a site at Yellowstone National Park shows that the
two volcanic areas have some similarity (Table 1).
Bacteriological studies of samples taken from soil with
and without Galdieria revealed that the number of
autotrophic bacteria growing by oxidation of FeSO , FeS
% #
(pyrite) and sulphur varied little between the two sites
(Table 2). The low numbers with pyrite as electron donor
are surprising because pyrite is present in the surface layer
of the rocks at these solfatara fields. When medium
containing galactose or mannose was inoculated with
material from the Galdieria-free site (2) we observed no
significant bacterial growth. In contrast, with inoculum
from site 1 we observed excellent heterotrophic growth of
Galdieria followed by growth of heterotrophic bacteria.
This implies that Galdieria is the primary heterotrophic
consumer in the tested habitat, but might excrete
compounds into the medium that are subsequently
consumed by heterotrophic bacteria. Using galactose or
mannose as the only carbon source, heterotrophic bacteria
27
seemed to be completely outcompeted by Galdieria under
the extreme conditions of pH 2±0 and a temperature of
50 °C.
At Pisciarelli and at nearby La Solfatara, organisms
meeting the description of Galdieria and Cyanidium were
found in soil and rock samples. The occurrence of Galdieria
and Cyanidium was limited to areas where the temperature
did not exceed 50–55 °C, as has been reported by Doemel
& Brock (1970). At higher temperatures only chemolitho-
autotrophic bacteria were found. Cyanidioschyzon merolae
was not observed in any of the samples, although this
species has been isolated from the same area by De Luca
et al. (1978). However, this extremely small and rare
species (De Luca et al., 1978) might easily have been
overlooked. Cells of the diatom Pinnularia were observed
in some samples, but were not abundant. Other eukaryotic
organisms were not found.
Cyanidium and Galdieria grew mainly on soil and rocks.
The underside of the rocks, in particular, showed a dark-
green layer of algae, even when these rocks were partly
embedded in mud. The colouring of the cells was always
dark-green to blue-green. Pale yellowish cells, typical of
heterotrophic Galdieria, were not observed. In addition
to cells attached to the surface of rocks (epilithic) and sand
grains (epipelic), algal mats were found under the rock
surface (cryptoendolithic) at various parts of Pisciarelli and
La Solfatara (Fig. 1). The algal layer was covered by a
glass-like greyish to yellowish layer 1–3 mm thick. The
analysis of this layer (Table 3) revealed that it consisted of
only two mineral phases : amorphous silica sinter as the
main constituent and trace amounts of amorphous ferrous
sulphide (FeS\nH O ; hydrotroilite), with very few crys-
#
talloid phases. The hydrotroilite will slowly convert to
pyrite (FeS ) and finally to FeOOH, which eventually
#
gives this layer a greyish-brown appearance. The next
layer consisted mainly of the algal biomass intermingled
with amorphous silica sinter and traces of elemental
sulphur. The underlying rock material was a porous,
relatively soft sandstone containing clay minerals and
elemental sulphur, but no iron. In some places the upper
layer was covered with elemental sulphur.
Transmission measurements of the upper silica sinter
layer taken from La Solfatara showed that on average
0±025–0±6 % of the daylight will reach the surface of the
algal layer. The lowest transmission was measured at
420 nm, increasing by 1 to 2 orders of magnitude above
500 nm (Fig. 2). However, at locations where the upper
layer is covered with elemental sulphur, far less light will
reach the algal layer. Unfortunately, the upper, sulphur-
covered layer was too brittle to permit the preparation of
pieces large enough for light measurements.
An ultrastructural examination of the algal layer
revealed that it consisted exclusively of algal cells with
the morphological characteristics of Galdieria and
Cyanidium (Fig. 3).
Because neither fungal hyphae nor heterotrophic bac-
teria were detected in the endolithic algal mats of La
Solfatara and Pisciarelli, Galdieria is without competitors
W. Gross et al. 28

Table 1. Physical and chemical parameters of various parts of the site at Pisciarelli, Italy compared with those of a site at Yellowstone
National Park, Montana

Sample Pond Spring A Spring B Muda Sandstonea Yellowstoneb

Temperature (°C) 23 92 92 30 47 40–55

pH 2±45 2±1 2±5 2±4 2±4 2–3
Conductivity (mS cm−") 20±0 19±8 7±6 7±7 22±2 0±9–2±1
DOC 11 9±2 12 240c 120c 7–14
SO #− – 3800 – 560c – 113–528
%
PO $− 32 43 27 41c 14c 8–0±8
%
Silica 5±7 144±0 71±4 0±48c 2±4c 220–270
Lead – ! 0±01 – 1±97c – –
Arsenic – 0±067 – 0±072c – 0±06–1±2
Cadmium – ! 0±001 – ! 0±001 – –
Mercury – ! 0±001 – ! 0±001 – 0±07–0±4

All concentrations are given in mg l−" (mg kg−" dry weight for soil and rock samples).
DOC, dissolved organic carbon.
a Aqueous extract (100 g dry weight}500 ml distilled water).
b Taken from Brock (1978).
c Per kg dry weight.

Table 2. Enumeration of auto- and heterotrophic bacteria and Table 3. Elemental composition (atomic %) of the upper rock
Galdieria from two different sites after incubation for 3 weeks in layer, the layer with endoliths, and the sandstone from the site at
the dark on various electron donors or carbon sources La Solfatara

Site 1, Galdieria-associated Site 2, Galdieria-free Element Upper layer Algal layer Sandstone

Electron Autotrophic Autotrophic Heterotrophic O 54±0 46±6 57±5

donor bacteria Galdieria bacteria bacteria C 0±0 17±5 0±0
Si 4±5 33±7 19±2
FeSO 11 000 ! 30 11 000 0 S 18±8 2±2 8±2
%
Sulphur 4600 ! 30 930 0 Al 1±3 0±0 10±5
FeS 140 ! 30 90 0 Fe 21±4 0±0 0±0
#
Mannose 0 2100a 0 90 K 0±0 0±0 4±5
Galactose 0 4600a 0 40

Values are expressed in cells per millilitre. The cell number (bacteria plus
algae) of the inoculum was 40. Controls (autotrophic medium) showed no
for organic substrates and obviously replaces fungi or
growth.
a All samples initially contained dividing Galdieria cells ; heterotrophic bacteria as the prime consumer. That this assumption is
bacteria slowly developed after about 3 weeks. realistic has been shown by an experiment where Galdieria

Fig. 1. Cross-section of a rock taken from a steep face at Pisciarelli, Italy, showing the endolithic algal layer (arrow). Scale bar represents
1 cm.
Endolithic growth of Galdieria
Fig. 2. Transmission spectrum of the mineral layer covering the
algal mat. Sample taken from La Solfatara, Italy. The four curves
represent four measurements at different areas of a single piece of
the upper layer.
Fig. 3. Scanning electron micrography of the algal layer adhering
to the subsurface of the upper mineral layer. Sample taken from
La Solfatara. Note the absence of fungal hyphae. Scale bar
represents 10 µm.
was fed with aqueous extracts obtained from cells of
Cyanidium or Galdieria. These extracts contained mainly
sucrose, glucose, galactose and glycerol. All compounds
were readily used by living Galdieria cells for hetero-
trophic growth. Cell growth ceased only when virtually
all of the sugar and glycerol had been consumed (Fig. 4).
The analysis of the soil and waters of the locations at La
Solfatara and Pisciarelli showed that sugar and sugar
alcohols were present in very low concentrations. The
highest concentration was found in soil samples where
glucose reached 15 µmol kg−" dry soil. Sucrose, galactose,
fructose and glycerol were even less concentrated. Other
sugars or sugar alcohols were not detectable. These results
indicate that the steady-state level of substrates for
Galdieria is very low.
29
Discussion
When we examined two volcanic sites near Naples we
identified the red alga Galdieria sulphuraria as the only
organism able to grow on organic compounds. No fungal
hyphae were detected by electron microscopy in algal
mats taken from Pisciarelli and La Solfatara. This contrasts
with the observations by Belly et al. (1973), who
consistently found Cyanidium together with the fungus
Dactylaria galloparva and the bacterium Bacillus coagulans
in algal mats on soils of the Yellowstone National Park,
Montana. The fungal biomass was directly proportional to
the algal biomass, while the density of the bacterium was
not. Therefore, Belly et al. (1973) suggested that under
these conditions the fungus is the prime consumer of any
organic material released from living or dead cells.
We did not detect heterotrophic bacteria either, al-
though chemolithotrophic bacteria were abundant. These
bacteria could benefit from the presence of Galdieria
because organic compounds with possible inhibitory
effects on the growth of the bacteria could be consumed
by the alga as has been observed for Acidiphilium sp.
(Harrison, 1984). However, although excretion by auto-
trophic bacteria can reach up to 20 % of the total carbon
fixed, the main product is glycolate (Schnaitman &
Lundgren, 1965 ; Cohen et al., 1979), a compound not
consumed by Galdieria (W. Gross, unpublished).
Although the natural habitat of the red algae Cyanidium
and Galdieria has been studied by various researchers
(Doemel & Brock, 1971 ; Pinto et al., 1994), cryptoendo-
lithic growth of these algae has never been reported.
Usually mats on soil and rocks have been described as
their habitat (Brock, 1978). Seckbach (1994) mentioned
that the algae were covered by a ‘ soft layer ’. Smith &
Brock (1973) reported that Cyanidium formed algal mats in
the soil covered by ‘ 3 mm of large granules of opaline
silica and quartz ’. In the volcanic area of Pozzuoli we
found that the cryptoendolithic occurrence of Cyanidium}
Galdieria is widespread, although not apparent at first
sight. The cryptoendolithic growth is most probably a
strategy to avoid desiccation, as reported for hot semi-
arid lands and deserts (Hoffmann, 1989 ; Bell, 1993)
as well as for arctic regions (Friedmann, 1982). The
upper layer of silica sinter is glass-like and shows only a
few cracks or fissures, thereby reducing evaporation
considerably. The occurrence of a distinct surface layer or
crust has rarely been reported in connection with
cryptoendolithic algae (Pentecost, 1978 ; Wessels & Bu$ del,
1995). This surface layer may cause problems by reducing
the available light for photosynthesis. Our transmission
measurements indicate that less than 1 % of the daylight is
available for photosynthesis at the surface of the algal
layer. A similar attenuation of light by the minerals has
been reported for cyanobacteria in carbonate deposits
(Pentecost, 1978). The low transmission rate of blue light
(350–450 nm) is counterbalanced by the presence of
phycocyanin in the alga with an absorption maximum at
520 nm (Gross & Schnarrenberger, 1995). Therefore, it
W. Gross et al.
Fig. 4. Heterotrophic growth of Galdieria sulphuraria (strain 002) supplied with an acidic extract of autotrophically grown Galdieria cells.
The composition and concentration of sugars and polyols in the medium (HPLC chromatogram) at the beginning (left) and end (right) of
the experiment are shown in the inserts.
can be assumed that at least limited photosynthesis by the
algae is possible. It is not clear how the algae colonize their
endolithic habitat. The cells could simply be washed into
the rock through minute cracks in the upper silica layer.
This would imply that the cells can somehow move
laterally after penetrating the rock. It is also possible that
the upper layer is formed after the cells have colonized the
rock surface. That the latter possibility is realistic is shown
by an experiment where a sandstone taken from Pisciarelli
was inoculated with Galdieria and supplied with high
concentrations of metasilicate. When the algae had
proliferated after a few days, the algal layer of the silica-
treated sample was covered by a soft, translucent deposit
of amorphous silica sinter. After about 4 weeks this silica
layer had become about 1 mm thick and the algae were
still dark blue-green in colour. After about 10 weeks the
cells gradually lost their pigmentation. The concentration
of metasilicate provided was about 20 times higher than is
usually found in hot acidic springs in order to speed up
the formation of a crust, which might have caused the
deterioration of the cells after 10 weeks. In the control no
silica layer developed but the algal growth was com-
parable.
In the natural habitat the gradual darkening of the silica
layer due to the formation of pyrite and FeOOH may
eventually lead to insufficient light penetration and
thereby deterioration of the algal layer. The organic
substances released by dead cells in the lower part of the
algal layer may not be sufficient for active growth. The
organic material could, however, be used by the cells to
maintain cell viability for longer periods. This has also
been suggested by Starks et al. (1981) for soil algae and by
Brock (1978) for Cyanidium growing in soils. The fact that
we found only trace amounts of sugars or polyols in
30
Pisciarelli and La Solfatara, Italy, could be explained by the
slow release of these substances from living or dead cells
or by a potent uptake system in the algal cells. For the
algal mats, at least, the latter possibility seems most likely.
Acknowledgements
This work was supported in part by the Deutsche
Forschungsgemeinschaft. We thank Professor G. Pinto,
Dr S. Esposito, and Professor C. Rigano for their help
during field studies.
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