Algological Studies 126 47–64 Stuttgart, April 2008

On Chlorogloeopsis fritschii (Cyanophyta/

Cyanobacteria) from thermal springs in
Slovakia and from a saline lake in Tunisia

FRANTIŠEK HINDÁK

Institute of Botany, Slovak Academy of Sciences, Bratislava, Slovakia

With 23 figures

Abstract: The life cycle of Chlorogloeopsis fritschii (Cyanophyta/Cyanobacteria,

Nostocales/Stigonematales) was studied in cultures isolated from fields in Al-
lahabad, India, thermal springs in Slovakia, Central Europe and a saline lake in
Tunisia, Northern Africa. As in the genera Nostoc, Trichormus or Nodularia, in
C. fritschii all vegetative cells were able to transform into akinetes. Two types of
germination of akinetes were distinguished. One type represented akinetes under-
going equal cell division and hormogonia formed without heterocytes that devel-
oped later. The second type was quite different and is unique in this group of cyan-
ophytes. The akinetes divided asymmetrically into a smaller part to be transformed
into a heterocyte, and a bigger one which divided again into two equal parts as veg-
etative cells. No substantial morphological differences were found among cultures
of the species isolated from India, Slovakia or Tunisia, although the places of origin
are ecologically different and geographically quite distant.

Key words: Chlorogloeopsis fritschii, cultured strains, Cyanophyta/Cyanobacte-

ria, life cycles, Nostocales/Stigonematales, salt lakes in Tunisia, ther-
mal springs in Slovakia

Introduction
The nostocalean/stigonematalean cyanophyte Chlorogloeopsis fritschii
(MITRA) MITRA et PANDEY 1966 is well known thanks to the cultures iso-
lated by MITRA (1950) from soils of wheat fields and gardens in Allahabad,
India. However, the species has not been recorded in the field again (see
ANAGNOSTIDIS & KOMÁREK 1990, KOMÁREK & ANAGNOSTIDIS 1998). The
cultures were deposited at main algal collections, e.g. ATCC, PCC (both
see RIPPKA et al. 1979), SAG (SCHLÖSSER 1994), CCALA (LUKAVSKÝ et
al. 1992) etc. and served as a suitable experimental object for many studies
(e. g. RIPPKA et al. 1979).

DOI: 10.1127/1864-1318/2008/0126-0047 1864-1318/08/0126-047 $ 4.50

© 2008 E. Schweizerbart’scheVerlagsbuchhandlung, D-70176 Stuttgart

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 48 F. HINDÁK

The species was recorded at thermal springs at Sklené Teplice in Cen-

tral Slovakia, but under the synonymous name Siphononema thermophila
HINDÁK (HINDÁK 1978). At the time of studying this new species, we were
not lucky enough to find heterocytes even in laboratory cultures, there-
fore we classified the species into the chroococcalean genus Siphononema
GEITLER 1925 (HINDÁK 1978, cf. KOMÁREK & ANAGNOSTIDIS 1998). Simi-
larly, no heterocytes were mentioned in the original description of Chlo-
rogloea fritschii by MITRA in 1950, therefore the author of the species clas-
sified it into the chroococcalean family Entophysalidaceae GEITLER 1932
(see also DESIKACHARY 1959).
In Slovakia, the species was repeatedly found in thermal springs at
Sklené Teplice, and recently also in a cultivated sample collected from the
dry salt lake Chott-el-Djerib, Tunisia, the largest saline lake in the Sahara.
UHER et al. (2005) published Chlorogloeopsis cf. fritschii from wet stones in
a gorge in the National Park Slovak Paradise, Central Slovakia. However,
their pictures resemble developmental stages of some other nostocacean
cyanophytes rather than C. fritschii, especially because of the presence of
baeocytes and the absence of asymmetrically formed heterocytes in aki-
netes.
In this contribution, the life cycle of Chlorogloeopsis fritschii from cul-
tures isolated from ecologically different and geographically distant locali-
ties will be described under light microscopy and documented by drawings
and micrographs. For comparison, the life cycle of the type strain PCC 6912
isolated by MITRA from India will be illustrated.

Material and methods

Thermal springs in Sklené Teplice

The town of Sklené Teplice is situated in Central Slovakia, in the Štiavnické

pohorie Mts, between the towns Banská Štiavnica and Žiar nad Hronom.
The thermal waters are sulphuric gypsum and the original temperature of
the Josef well (Fig. 1) is 52.3 °C (some more data see HINDÁK 1978). The
investigated material was collected in 1975–2006. Strains HINDÁK 1975/134
(see HINDÁK 1978) and HINDÁK 1983/3 were isolated from the thermal
spring Joseph and HINDÁK 2005/10 from the thermal spring Vilma.
Cultures grew very well under standard laboratory conditions, i.e. at
temperature of about 25–30 °C and illumination by fluorescent tubes (see
HINDÁK 1978, HINDÁK & ŠMARDA 2006).

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 Chlorogloeopsis fritschii from thermal springs 49

Fig. 1. Thermal springs at the Sklené Teplice, Central Slovakia;

photo A. HINDÁKOVÁ, May 10, 2006.

The dry salt lake Chott–el–Djerib

Strains were isolated from a sample from the sand bank of an excava-
tion pool in the dry saline lake Chott–el–Djerib, Central Tunisia (Fig. 2; cf.
also HINDÁK & ŠMARDA 2006), collected on January 3, 2006 by Prof. Jozef
HALGOŠ, Dept. of Zoology, Comenius University, Bratislava. In the fresh
sample no cyanophytes and algae were found probably because of heavy
rains in December 2005.
In the laboratory the sample was transferred to Petri dishes with a salt
medium (Red Sea Salt used for aquaria) and put in a cultivation box. Within
a couple of weeks of cultivation the first pioneering algae and cyanophytes
appeared, mostly representing the genera Dunaliella TEODORESCU, Aph-
anothece NÄGELI, Asterocapsa CHU, Pseudanabaena LAUTERBORN, Nostoc
VAUCHER ex BORNET et FLAHAUT and Chlorogloeopsis MITRA et PANDEY.
Some of these were successfully isolated into unialgal cultures. Strains of
Chlorogloeopsis fritschii (MITRA) MITRA et PANDEY grew vigorously under
standard laboratory conditions in liquid or on agarized marine as well as in
freshwater media (water or biphasic BBM, see HINDÁK & ŠMARDA 2006).
The colour of mature colonies in the salt medium was brownish, while in
freshwater media it was bright to dark green.

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 50 F. HINDÁK

Fig. 2. An excavation pool in the dry saline lake Chott-el-Djerib, Central Tunisia;
photo J. HALGOŠ, Jan. 4, 2003.

Cultures

The type strain PCC 6912 isolated by MITRA was received from the Pasteur
Culture Collection, Paris (PCC No. 6912, see RIPPKA et al. 1979). The stud-
ied cultures HINDÁK 2005/10 and HINDÁK 2006/4 are deposited in the Cul-
ture Collection of Autotrophic Organisms (CCALA) at Třeboň, the Czech
Republic.
A Leitz Diaplan microscope equipped with a Wild Photoautomat MPS
45 was used for LM investigations.

Results and discussion

Our observations on the life cycle of all investigated cultures are in good
agreement with previous data on Chlorogloeopsis fritschii from India
(MITRA 1950, MITRA & PANDEY 1966, DESIKACHARY 1959) and studies of
RIPPKA et al. (1979). No substantial morphological differences were found
among cultures isolated from India (Figs 3–6), Slovakia (Figs 7–16) and Tu-
nisia (Figs 17–22).
In natural material the species is not easily recognised because colo-
nies are not well developed or have disintegrated into small aggregates or

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 Chlorogloeopsis fritschii from thermal springs 51

Fig. 3. Chlorogloeopsis fritschii, type strain (PCC 6912) isolated by MITRA from
India; filaments; Scale bar 10 µm.

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 52 F. HINDÁK

Fig. 4. Chlorogloeopsis fritschii, type strain (PCC 6912) isolated by MITRA from
India; subcolonies; Scale bar 10 µm.

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 Chlorogloeopsis fritschii from thermal springs 53

Fig. 5. Chlorogloeopsis fritschii, type strain (PCC 6912) isolated by MITRA from
India; filaments and germination of akinetes. [Scale bar = 10 µm].

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 54 F. HINDÁK

Fig. 6. Chlorogloeopsis fritschii, type strain (PCC 6912) isolated by MITRA from
India; different stages of life cycle: germination of akinetes, hormogonia and fila-
ments. [Scale bar = 10 µm].

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 Chlorogloeopsis fritschii from thermal springs 55

cells, reminiscent of the genera Chroococcidiopsis GEITLER, Chroococcus

NÄGELI or Nostoc BORNET et FLAHAUT. However, under laboratory condi-
tions the cells divide intensively and form large colonies (Figs 3, 4, 8, 11, 12,
17, 18). The colonies are conspicuous by their firm and compact margins
but their size and form are ill defined, mostly elongate and lobed, sausage–
like or irregularly globose, with a narrow gelatinous sheath that is usually
poorly visible (Figs 3–6, 9, cf. also HINDÁK 1978, Fig. 11). The colonies are
composed of densely clustered and irregular cell packets or uniseriate to
multiseriate filaments arranged in vertical or horizontal rows, diffluent on
the margin particularly in old colonies.
Young hormogonia (Figs 6, 7, 10, 12, 13, 19) form uniseriate, short, chain–
like filaments, slightly motile, with shortly cylindrical to spherical or sub-
spherical cells, initially without heterocytes or during later development
with one apical or intercalary positioned heterocyte. From the beginning,
the cells divide in one direction but commonly after reaching 5–7 cells
growth is in two or more directions, resulting in packet-like colonies or
bunchy and maize spike-resembling configurations (cf. also HINDÁK 1978).
Adult cells are more or less spherical, after division subspherical or
asymmetrically Chroococcus-like when in packets. Depending on age, they
are bright to dark yellow green, blue green or olive green in colour, (2)–3–
6–(8) µm in diameter, sometimes with fine granules particularly in the cen-
tral part of the cell. The mucilage envelope around the cells is rarely con-
centrically layered (Fig. 10), usually it is confluent and indistinct except in
old cultures (Figs 5, 9), 5–7 µm wide, hyaline, only rare yellowish to brown.
Aerotopes were not observed.
As in representatives of some nostocacean genera [e. g. Nostoc, Trichor-
mus (RALFS ex BORNET et FLAHAUT) ANAGNOSTIDIS et KOMÁREK, Nodu-
laria MERT. ex BORNET et FLAHAUT – see STARMACH 1966, HINDÁK 2001,
KOMÁREK et al. 2003], all vegetative cells in C. fritschii are able to transform
into akinetes (Figs 8, 21, 22). Germination of akinetes does not require a
special period of dormancy, the akinetes are able to germinate both in ma-
ture cultures, and after transfer of an old culture to fresh nutrient media.
Akinetes are more or less spherical, (6)–8–12 µm in diameter, with hyaline
or rare brown cell wall.
Two types of germination of akinetes can be distinguished in Chlorogloe-
opsis fritschii (cf. Fig. 6). One type represents the akinetes, in which the cell
division results in two equal parts and hormogonia are formed without het-
erocytes that are developed later (i.e. after liberation of the hormogonium
from the akinete sheath). The second type is quite different and perhaps
unique in this group of cyanophytes. The akinetes divide asymmetrically
into two parts: one smaller part is transformed into a heterocyte and the
bigger one divides again into two equal parts as vegetative cells (Figs 5,
6, 15, 21). Thus hormogonia or filaments arising from such akinetes have
a distinct terminal heterocyte (Figs 3, 8,10, 21, 22) as in the genus Nostoc

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 56 F. HINDÁK

Fig. 7. Chlorogloeopsis fritschii, strain HINDÁK 1983/3 isolated from the thermal
spring Joseph at Sklené Teplice, Slovakia; hormogonia, filaments and germination
of akinetes. [Scale bar = 10 µm].

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 Chlorogloeopsis fritschii from thermal springs 57

Fig. 8. Chlorogloeopsis fritschii, strain HINDÁK 1983/3 isolated from the thermal
spring Joseph at Sklené Teplice, Slovakia; germination of akinetes, filaments and
the shape of a colony at small magnification (upper right). [Scale bar = 10 µm].

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 58 F. HINDÁK

Fig. 9. Chlorogloeopsis fritschii, strain HINDÁK 1983/3 isolated from the thermal
spring Joseph at Sklené Teplice, Slovakia; uniseriate hormogonia, multiseriate fila-
ments and germination of akinetes. [Scale bar = 10 µm].

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 Chlorogloeopsis fritschii from thermal springs 59

Fig. 10. Chlorogloeopsis fritschii, strain HINDÁK 1979/22 isolated from the thermal
spring Joseph at Sklené Teplice, Slovakia; germination of akinetes, uniseriate hor-
mogonia and multiseriate filaments. [Scale bar = 10 µm].

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 60 F. HINDÁK

Figs 11, 12. Chlorogloeopsis fritschii, strain HINDÁK 2005/10 isolated from the
thermal spring Vilma at Sklené Teplice, Slovakia: 11 – shape of a colony; 12 – part
of a colony with hormogonia.

Figs 13, 14. Chlorogloeopsis fritschii, strain HINDÁK 2005/10 isolated from the
thermal spring Vilma at Sklené Teplice, Slovakia; uniseriate to multiseriate hormo-
gonia.

Figs 15, 16. Chlorogloeopsis fritschii, strain HINDÁK 2005/10, isolated from the
thermal spring Vilma at Sklené Teplice, Slovakia; germination of akinetes, h – het-
erocytes.

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 Chlorogloeopsis fritschii from thermal springs 61

Figs 17, 18. Chlorogloeopsis fritschii strain HINDÁK 2006/4 isolated from a saline
lake Chott-el-Djerib, Tunisia; shape of a colony.

Figs 19, 20. Chlorogloeopsis fritschii strain HINDÁK 2006/4 isolated from a saline
lake Chott-el-Djerib, Tunisia; uniseriate hormogonia and multiseriate filaments.

Figs 21, 22. Chlorogloeopsis fritschii, strain HINDÁK 2006/4 isolated from a saline
lake Chott-el-Djerib, Tunisia; germination of akinetes, h – heterocytes.

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 62 F. HINDÁK

Fig. 23. Scheme of the life cycle in genera Chlorogloeopsis and Fischerella; after
RIPPKA et al. (1979).

(GEITLER 1932, 1960, HOLLERBACH et al. 1953, DESIKACHARY 1959, KON-

DRATEVA 1968, ANAGNOSTIDIS & KOMÁREK 1990, KOMÁREK & ANAGNOS-
TIDIS 1989, 1998). Heterocytes are mostly smaller than vegetative cells, with
thick cell walls. The shape of terminal heterocytes is broadly conical, sub-
spherical to spherical, intercalary heterocytes are subspherical to spherical.
As seen from the data of its occurrence (India, Slovakia, Tunisia), Chlo-
rogloeopsis fritschii is probably a widespread species that is difficult to
identify in field material, but not in laboratory cultivated material or sub-
cultures. It can be considered a soil organism that seems to prefer higher
temperatures and also higher amounts of minerals. This is evident from
the occurrence of this species in thermal springs in Central Slovakia with
a temperature of about 50 °C and high concentrations of minerals (min-
eral waters), and in a hypersaline lake in northern Sahara in Tunisia. Be-
cause the cultures of C. fritschii isolated from a saline lake grow very well
in freshwater media, we can assume that this cyanophyte is not obligatory
halophytic.

Taxonomic conclusions
According to literary data and our studies of cultures isolated from India,
Slovakia and Tunisia, we concur with ANAGNOSTIDIS & KOMÁREK (1990)
that the genus Chlorogloeopsis with the single species C. fritschii is a well-
defined cyanophyte that deserves to be classified in a special family Chlo-
rogloeocapsaceae. On one hand, the ability to transform all vegetative cells
into akinetes is similar to some genera in the family Nostocaceae, e.g. Tri-
chormus, Nodularia or Nostoc. On the other hand, its life cycle resembles

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 Chlorogloeopsis fritschii from thermal springs 63

the genus Fischerella GOMONT in the family Stigonemataceae as is shown

by the scheme of RIPPKA et al. (1979).
Asymmetrical division of cells resulting in a small heterocyte and a big
vegetative cell is a typical feature of C. fritschii which has not been found in
the nostocacean/stigonematalean cyanophytes. It resembles the formation
of heterocytes in the planktonic genus Anabaenopsis (WOŁOSZ.) V. MILL.,
in which, however, the pattern is quite different (cf. KOMÁREK & ANAGNOS-
TIDIS 1989, HINDÁK 2001).
The proposed taxonomic classification of C. fritschii is as follows:
Order: Stigonematales GEITLER 1925
Family: Chlorogloeocapsaceae (MITRA) MITRA et PANDEY 1966
Genus: Chlorogloeopsis MITRA et PANDEY 1966
Species: Chlorogloeopsis fritschii (MITRA) MITRA et PANDEY 1966
Basionym: Chlorogloea fritschii MITRA 1950
Synonym: Siphononema thermophila HINDÁK 1978; incl.
O c c u r r e n c e : soils in India (1950), thermal springs in Slovakia (1978 and
this report) and a saline lake in Tunisia (this report).

Acknowledgements

The author expresses his sincere thanks to Prof. Jozef HALGOŠ, Bratislava, for sam-
pling of material from Tunisia in 2006 and to Dr. Hans J. SLUIMAN, Edinburgh, for
correcting the English. This study is a part of the VEGA projects Nos 2/4033/04
and 2/7069/27 supported by the Slovak Academy of Sciences.

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Manuscript received December 29, 2006, accepted September 23, 2007.

The author’s address:

Prof. RNDr. FRANTIŠEK HINDÁK, DSC.

Slovak Academy of Sciences
Institute of Botany
Dúbravská cesta 14
SK–84523 Bratislava, Slovakia
e-mail: frantisek.hindak@savba.sk

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