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Cyanobacteria in ambient springs

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DOI: 10.1007/s10531-015-0884-x

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Cyanobacteria in ambient springs

Marco Cantonati, Jiří Komárek &

Gustavo Montejano

Biodiversity and Conservation

ISSN 0960-3115
Volume 24
Number 4

Biodivers Conserv (2015) 24:865-888

DOI 10.1007/s10531-015-0884-x

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 Author's personal copy
Biodivers Conserv (2015) 24:865–888
DOI 10.1007/s10531-015-0884-x

REVIEW PAPER

Cyanobacteria in ambient springs

Marco Cantonati • Jiřı́ Komárek • Gustavo Montejano

Received: 24 August 2014 / Revised: 30 January 2015 / Accepted: 12 February 2015 /

Published online: 21 February 2015
Ó Springer Science+Business Media Dordrecht 2015

Abstract Although neglected for a long time by freshwater-ecology research, springs are
very important habitats for biodiversity conservation. They are multiple ecotones, and are
characterized by a remarkable variety of environmental conditions (e.g., from highly-
shaded to UV exposed, from permanent discharge to intermittent flow, from still water to
strong currents, from extremely-soft to carbonate-precipitating water, etc.). Moreover,
springs are often amongst the last high-integrity, oligotrophic freshwater habitats in
densely populated areas. Because of the high quality of their waters, the main impact
affecting springs is capturing and water diversion. Climate-change driven reduction in
precipitations in many areas is likely to determine an aggravation of this impact. It is thus
important to document the rich and peculiar biodiversity of springs, also to establish
reference conditions for bioassessment methods. Especially in non-acidic springs with
running water, and coarse lithic substrata, cyanobacteria are often one of the most taxa-rich
and abundant groups of photoautotrophs. The relatively-scarce information available in the
literature is mostly referred to similar habitats, and not to spring habitats in the narrower
sense. Papers dealing with the cyanobacteria of ambient springheads (=eucrenal) world-
wide are still very rare. These were reviewed separating ambient springs in temperate and

Communicated by Anurag Chaurasia.

M. Cantonati (&)
Museo delle Scienze - MUSE, Limnology and Phycology Research Unit, Corso del Lavoro e della
Scienza 3, 38123 Trento, Italy
e-mail: marco.cantonati@muse.it

J. Komárek
Faculty of Biological Sciences, University of South Bohemia, České Budějovice, Czech Republic

J. Komárek
Institute of Botany AS CR, Dukelská 135, 379 18 Třeboň, Czech Republic

G. Montejano
Facultad de Ciencias, Laboratorio de Ficologı́a, UNAM-Universidad Nacional Autónoma de México,
Avenida Universidad 3000, Ciudad Universitaria, 04510 Coyoacán, Federal District of México City,
México

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866 Biodivers Conserv (2015) 24:865–888

warm climate, and with special attention to key species, to cyanobacterial strategies al-
lowing survival in oligotrophic headwaters (e.g., nitrogen fixation, phosphatases, anti-UV
compounds, etc.), and to distribution patterns. The review also hopes to bolster new
interest and research on this topic, and suggests some promising research directions.

Keywords Springs
Cyanoprokaryotes
Oligotrophy
Radiation
Nitrogen
Discharge

fluctuations

Introduction

Cyanobacteria are well adapted to oligotrophic environments, and are generally among the
dominant primary producers in spring habitats. Cyanobacteria or cyanoprokaryotes (Komárek
and Golubić 1990) are the most ancient oxygenic photoautotrophs, colonize a wide variety of
habitats including extreme ones, are important primary producers at the global scale (e.g.,
Sciuto and Moro 2015). Despite the many problems related to the application of species
concepts to these asexual prokaryotes, cyanobacteria include a remarkable diversity of mor-
photypes and genotypes (including cryptic species and genera) (Dvořák et al. 2014, 2015).
Springs are extremely diverse ecotones, which can be classified according to different
criteria derived from hydrology, geology, hydrochemistry, water temperature, ecology, and
human use (Glazier 2009). Various cyanobacterial taxa, which are very diverse in de-
pendence on environmental conditions, are important in the ecological system of springs,
but are also very variable in springs of various types. Springs are extremely endangered in
populated countries, and the study of the dominating cyanobacterial microflora is important
in relation to the control and conservation of water sources.
When several organism groups are used for spring classification (e.g., Spitale et al. 2012a),
cross-taxon congruency is weak (as shown by Nascimbene et al. 2011 for crenic photoau-
totrophs including cyanobacteria and algae). Thus spring conservation plans should not use
one group as a surrogate but consider different groups of organisms. Springs characterized by
a temperature approaching the mean annual air temperature (MAAT) in the drainage basin,
have traditionally been called cold springs, but they should be renamed ambient springs,
because cold springs should be those that have temperatures below the MAAT. By contrast,
thermal springs have water temperatures clearly higher than MAAT, while hot springs have
temperatures exceeding human-body temperature (Glazier 2009). Using a set of available
estimates, it was concluded that there should be worldwide on land (not considering
Antarctica) probably more than four ambient springs per km2 (global total [57 9 106).
Thermal springs would be much rarer: global total [105 (Glazier 2009). Most springs show
hydrochemical characteristics consistent with the general lithological classification of the
aquifer. However, several cases of springs with ‘‘special’’ hydrochemistry, characterized by
high concentrations of one or more ions and/or elements (e.g., sulphate, sulphide, iron,
arsenicum, copper etc.) also exist: these are called mineral springs. Most hot springs are also
mineral springs (e.g., Castenholz 1969).
Spring habitats are multiple ecotones (subaerial-submersed, springhead-spring stream,
groundwater-surface waters). They are of special relevance for biodiversity conservation
because of the high diversity they host. However, studies dealing with different organism
groups showed that high species numbers are generally found not in single springs but in
regional pools including sufficient numbers of sources representative of the different spring
types and geological substrata (Nascimbene et al. 2011; Cantonati et al. 2012a). Spring

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Biodivers Conserv (2015) 24:865–888 867

assemblages moreover include high proportions of rare and red list taxa, and species that
are too sensitive to colonise other freshwater habitats that are more impacted in densely
populated and highly exploited geographic areas [named least-impaired habitat relicts
(LIHRe) by Cantonati et al. 2012a]. The main cause of this high diversity is likely to be the
remarkable variety of environmental conditions that the individual springs can represent
(e.g., from highly-shaded to UV exposed, from highly-stable, permanent discharge to
markedly-fluctuating or even intermittent flow, from still water to strong currents, from
extremely-soft to petrifying water etc.).
Starting from the three basic spring types (rheocrenes = flowing springs, helocre-
nes = seepages, limnocrenes = pool springs), several others were added, the main ones
being hygropetric springs or rock-face seepages, and the so called tufa springs, that should
more appropriately be named spring-associated-limestone (SAL) sources (Sanders et al.
2010). Spring types can be identified and characterized also by means of benthic algae
(Cantonati et al. 2012b).
Springs are very heterogeneous not only at the scale of habitat but also at the scale of
microhabitat, i.e. the different substrata within a spring (the most common ones are lithic
material, bryophytes, surface sediment, and organic debris). Springs are thus often said to
be microhabitat mosaics (Hájek et al. 2011; Hájková et al. 2011; Cantonati et al. 2012a).
While cyanobacteria in hot springs have been more in depth studied and even recent
review papers are available (e.g., Ward et al. 2012), cyanoprokaryotes in ambient springs
are a neglected topic, in spite of ambient springs being numerous and widespread and of
the high ecological and evolutionary interest of cyanobacteria. When they are considered,
they are often studied in the frame of all benthic algae (this includes all prokaryotic and
eukaryotic algae that are macroscopic or form thalli or generate colorations visible by the
naked eye; Cantonati et al. 2012b).
The relatively-scarce information available in the literature is mostly referred to similar
habitats and not to the springs sensu stricto. There is, for instance, typically confusion
between springs and spring-fed streams, and not rarely it is virtually impossible to dis-
tinguish the data relative to springs from those referred to the running-water systems
studied (e.g., Hu and Xie 2013). Moreover, spring studies including cyanoprokaryotes are
rare (e.g., Dell’Uomo 1975), and were mainly carried out in the framework of researches
focused on other topics (e.g., hygropetric habitat, Golubić 1967; mountain streams, Kann
1978). Some of the early studies on periphyton algae in springs indicated that these
environments host no specific spring species (e.g., Whitford 1956; Whitford and Schu-
macher 1963), while a more recent investigation described a new red-alga species sup-
posed to be exclusive of spring habitats (Vis and Sheath 1996).
A literature search of the Institute for Scientific Information (ISI) Web of Knowledge
(publication period: all years, updated 9 August 2014, databases = SCI-EXPANDED,
SSCI, A and HCI) for the keyword ‘‘spring*’’, refining the search to the web-of-science
category ‘‘marine freshwater biology’’, and then searching within the results for ‘‘cyano*’’
yielded 515 articles. However, direct inspection of titles and abstracts revealed that the
majority of these papers (490) were wrong matches (e.g., ‘‘spring’’ was the season etc.).
Thirteen were devoted to cyanobacteria in hot springs, while only twelve were really
dealing with cyanoprokaryotes in ambient springs.
The aim of this paper is to review the sparse and limited literature available on
cyanoprokaryotes in ambient springs to stimulate further research, and suggesting
promising research directions. The focus is on the biodiversity of cyanobacteria in ambient
springs, on their distribution in the different spring types, on cyanobacterial adaptations to

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868 Biodivers Conserv (2015) 24:865–888

these special environments, and on the conservation status and impacts menacing this
component of ambient-spring biodiversity.

Cyanobacteria in ambient springs: diversity; main characteristics, rare taxa

(temperate climate) (Fig. 1a–l)

The number of cyanoprokaryote taxa found in ambient springs is about 15-20 taxa every 10
springs investigated, and the number of cyanoprokaryote taxa per spring varies between 0 and
10 with an average of 4 (Table 1). A prevalence of coccoid types, with proportions typically
between 50 and 60 %, is evident. The most frequent and abundant genera are Chamaesiphon
(Fig. 1a, b, j), Tapinothrix (Fig. 1c), Phormidium (Figs. 1e, f), Leptolyngbya, Heteroleibleinia
(Fig. 1d), Pleurocapsa. Rare species include: Chamaesiphon amethystinus (Fig. 1j), Xeno-
tholos kerneri (Fig. 1k), Cyanodermatium sp., Myochrotes myochrous (Fig. 1i), Petalonema
spp., Tapinothrix bornetii (type species of the genus Tapinothrix), Gloeothece rupestris. The
most commonly studied spring type are rheocrenes, and consistently investigations focused
mostly on the epilithon (the most common substratum in this spring type; Table 1), more rarely
on the epibryon (Hašler and Poulı́čková 2005; Cantonati 2008).
Some species are recorded frequently from springs (e.g., Chamaesiphon amethystinus,
Starmach 1929; C. confervicola-type species of the genus Chamaesiphon Golubić 1967; Ho-
moeothrix juliana, Komárek and Kann 1973), but none of them seems to thrive in spring
habitats only. In a longitudinal zonation study, Spitale et al. (2012b) found several species in the
eucrenon (spring head sensu stricto) (Chamaesiphon confervicola, C. investiens var. roseus,
Cyanodermatium sp.), whereas Chamaesiphon fuscus, C. polonicus, and Leptolyngbya frigida
were more abundant in epirhithral downstream sections (spring-fed stream).

Cyanobacteria in the different spring types

Helocrenic and limnocrenic springs

Early studies focusing on epilithon (Cantonati et al. 1996) noted that cyanoprokaryotes
were scarce or absent in helocrenic and limnocrenic springs in which muddy substrata
prevail, and rocky material is often unavailable. Cantonati et al. (1996) proposed the
following potential causes for the lack of cyanobacteria in these spring types: absence of
appropriate substratum for epilithic taxa, acidic conditions and high values of free carbon
dioxide often found in these spring types. Later studies confirmed this observation (Can-
tonati 2008; Cantonati et al. 2012b).
However, a cyanobacterial microflora of richness comparable to that usually found in
epilithon studies could be illustrated by Hašler and Poulı́čková (2005), who studied epi-
bryon cyanoprokaryotes in Western Carpathian helocrenic springs. Cyanobacteria, how-
ever, were absent from acidic springs in this study as well, and were documented from
circumneutral and alkaline springs only.

Rheocrenic springs on siliceous substratum

There is a clear difference between the cyanoprokaryote assemblages in flowing springs on

siliceous and carbonate substratum. Even taxa vicariant in springs on the two substrata

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Biodivers Conserv (2015) 24:865–888 869

Fig. 1 Some representative, characteristic, and rare cyanoprokaryotes from temperate-climate ambient-
temperature springs. a Chamaesiphon confervicola, b Chamaesiphon starmachii, c Tapinothrix janthina
(scale bar = 3 lm), d Heteroleibleinia purpurascens, e Phormidium favosum, f Phormidium retzii (? violet
apex of P. insigne filament); g Tolypothrix penicillata (scale bar = 150 lm), h Rivularia biasolettiana
(SAL spring) (scale bar = 50 lm), i Myochrotes myochrous (scale bar = 20 lm), j Chamaesiphon
amethystinus, k Xenotholos kerneri (scale bar = 15 lm), l Schizothrix tinctoria (scale bar = 2 lm). If not
otherwise stated, the scale bars equal 5 lm

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 Table 1 Cyanobacteria in ambient springs: synthesis of literature data
870

Geographic No. of Types of No. %C. %F. %H. %Cy. No. of Main species Notes and taxa of special References
location springs springs of taxa per interest

123
studied considered; taxa spring
elevation found average
range (m (min–
a.s.l.); max)
microhabitat
considered

South- 55 Rh-si, Rh-ca, 83 51 36 13 73 4(0–11) T. varians (ca), Chamaesiphon Seven spring types identified Cantonati et al.
eastern He-si, Hy, polonicus, Phormidium by means of fuzzy (2012a,
Alps: SAL, Ir, favosum, Chamaesiphon clustering of benthic algal 2012b,
Trentino RG; subglobosus, Tapinothrix assemblages: Half of the 2012c)—
250-2739; janthina (si), Pleurocapsa indicator taxa identified by CRENODAT
El(Ss) minor, Heteroleibleinia IndVal analysis were only
fontana, H. purpurascens, cyanoprokaryotes.
Chamaesiphon starmachii, Chamaesiphon
Leptolyngbia spp., amethystinus, Xenotholos
Tapinothrix crustacea, kerneri, Cyanodermatium
Phormidium incrustatum sp., Ammatoidea normannii,
Petalonema densum
South- 16 Rh-ca, Li, Hy, 36 50 36 14 73 3(0–8) Tapinothrix varians, Prevalence of cyanobacteria Cantonati
Author's personal copy

eastern Ir; Chamaesiphon polonicus, in Rh, of green algae in Li. (2008)—

Alps: 511-1979; Chlorogloea sp. small cells, Epilithon: higher numbers springs only
Dolomiti El and Phormidium favosum, of taxa and diversity than
Bellunesi Eb(Ss) Xenotholos kerneri, epibryon. Very-reduced
National Chamaesiphon subglobosus diversity in Ir. Ammatoidea
Park normannii, Geitleribactron
periphyticum, Xenotholos
kerneri, Tapinothrix
bornetii, Tolypothrix
pulvinata
Biodivers Conserv (2015) 24:865–888
 Table 1 continued
Geographic location No. of Types of No. %C. %F. %H. %Cy. No. of Main species Notes and taxa of special References
springs springs of taxa per interest
studied considered; taxa spring
elevation found average
range (m (min–
a.s.l.); max)
microhabitat
considered

South-eastern Alps: 19 Rh-si, Rh-ca, 40 52 38 10 100 6 (0–10) Tapinothrix janthina (si), No cyanoprokaryotes in He. Cantonati
Adamello-Brenta He, Hy; T. varians (ca), Maximum N. of taxa in Hy. et al.
Biodivers Conserv (2015) 24:865–888

Nature Park, River 1073-2130; Phormidium favosum, Chamaesiphon (1996)

Sarca catchment El,Eb(Ss) Chamaesiphon amethystinus, C. investiens
polonicus, C. var. roseus, Xenotholos
confervicola, C. kerneri, Phormidium
minutus, Leptolyngbya tinctorium,
perforans (ca) Cyanodermatium sp.
Central Alps: 23 Rh-si, Rh-ca, 62 61 28 11 77 – Tapinothrix varians (ca), No soft-algae covers in Gesierich
Vorarlberg, Austria Hy, SAL, Chamaesiphon sulphur spring. and
Ir; polonicus, Phormidium Chamaesiphon Kofler
572-1840; favosum, Leptolyngbya amethystinus, Gloeothece (2010)C
El perforans (ca) linearis, Rivularia
Author's personal copy

periodica, Schizothrix
tinctoria, Myochrotes
myochrous
Central Alps: Ötztal, 5 Rh, Li; El 12 42 33 25 63 7(3–12) Tapinothrix varians, Ammatoidea normannii Gesierich
Tyrol, Austria. Tolypothrix penicillata, and Rott
UNESCO Biosphere Chamaesiphon (2004)—
Reserve ‘‘Gurgler polonicus, Phormidium springs
Kamm’’, Catchment favosum only
of the Rotmoos
glacial stream
871

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 Table 1 continued
872

Geographic No. of Types of No. %C. %F. %H. %Cy. No. of Main species Notes and taxa of special References
location springs springs of taxa per interest

123
studied considered; taxa spring
elevation found average
range (m (min–
a.s.l.); max)
microhabitat
considered

Surroundings of 4 F; El 15 93 7 0 100 4(2–8) Pleurocapsa cf. aurantiaca, P. Gloeothece rupestris, Uher et al.
the town of minor, Chamaesiphon Rhabdoderma irregulare (2001)
Brno, polonicus
Moravia,
Czech
Republic
South-eastern 1 Rh(Li)-si; 12 42 42 16 48 12 Chamaesiphon polonicus, Aphanocapsa pulchra Dell’Uomo
Alps, 1100; Phormidium favosum, (1975)
Anterselva El ? Ep Tapinothrix janthina
Valley,
Autonomous
Province of
Bozen
Author's personal copy

Western 14 He; 370-905; 48 42 35 23 – 5(0–16) Aphanocapsa parietina, No cyanoprokaryotes Hašler and

Carpathians: Eb Aphanothece saxicola, obtained from acidic Poulı́čková
Bı́lé Karpaty Chamaesiphon polonicus, sites. Chroococcus (2005)
Mts & Chroococcus minor, minutus, Microchaete
Moravsko- Gloeothece palea, tenera, Nostoc commune,
Slezské Leptolyngbya boryana, N. microscopicum,
Beskydy Mts Nostoc commune, N. Trichormus variabilis
(Czech muscorum, Phormidium
Republic) retzii, P. tenuissimum
Biodivers Conserv (2015) 24:865–888
 Table 1 continued
Geographic No. of Types of springs No. %C. %F. %H. %Cy. No. of Main species Notes and taxa References
location springs considered; of taxa per of special
studied elevation range taxa spring interest
(m a.s.l.); found average
microhabitat (min–
considered max)

Pego-Oliva 6 Rh in arid 4 – 75 25 100 2–3 Calothrix parietina, IMPORTANT Cantoral-Uriza and Aboal-
marsh (East climate Homoeothrix juliana, NOTE: Only Sanjurjo (2010); Garcı́a and
Iberian Leptolyngbya macroalgae Aboal (2014); Garcı́a-
Peninsula angustissima, considered. Fernández (2014); Garcı́a-
Biodivers Conserv (2015) 24:865–888

Schizothrix Fernández and Aboal

fasciculata, (personal com.)
Tapinothrix crustacea
Pego-Oliva 2 ISP in arid 5 20 60 20 90 1–3 Calothrix pulvinata, Important Cantoral-Uriza and Aboal-
marsh (East climate Phormidium note: only Sanjurjo (2010); Garcı́a and
Iberian tergestinum, macroalgae Aboal (2014); Garcı́a-
Peninsula) Tapinothrix violacea considered Fernández (2014); Garcı́a-
Fernández and Aboal
(personal com.)
Volusia Blue 1 Rh-ca; El 15 27 73 0 58 15 Heteroleibleinia Stichosiphon Wehr (personal comm.)
Spring, Blue kuetzingii, willei,
Author's personal copy

Spring State Leptolyngbya spp., Xenotholos

Park, Orange Oscillatoria spp., kerneri
City, Florida, Phormidium cf. retzii
USA
873

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 Table 1 continued
874

Geographic No. of Types of No. %C. %F. %H. %Cy. No. of Main species Notes and taxa of special References
location springs springs of taxa per interest

123
studied considered; taxa spring
elevation found average
range (m (min–
a.s.l.); max)
microhabitat
considered

Springs, 30 Rh-ca, Su 105 31 60 9 33 Amphithrix sp., Calothrix sp., Whitford

Florida Lyngbya epiphytica, Lyngbya (1956)
State, kutzingii, Lyngbya
USA nordgardhii, Oscillatoria
splendida, Plectonema wollei,
Xenococcus sp. (hard-water
springs)
Panuco 14 Rh-ca, Li-ca; 234 54 36 10 23 Blennothrix ganeshii, Chamaecalyx calyculatus, C. Montejano
River 60-500 Chamaesiphon confervicola, swirenkoi, Chlorogloea et al.
basin, Cyanocystis mexicana, epiphytica, Hyella (2000);
central Heteroleibleinia spp., kalligrammos, Romeria sp., Carmona-
Mexico Homoeothrix juliana, Stichosiphon exiguus, S. Jiménez
Hydrococcus rivularis, Hyella sansibaricus, Xenotholos and
Author's personal copy

cf. fontana, Merismopedia huastecanus, Xenococcus Montejano

glauca, Pleurocapsa minor, bicudoi (1993)
Synplocastrum
muelleri,Stichosiphon
filamentosus, Xenococcus
willei, X. kerneri
Biodivers Conserv (2015) 24:865–888
 Table 1 continued
Geographic No. of Types of springs No. %C. %F. %H. %Cy. No. of Main species Notes and taxa of special References
location springs considered; of taxa per interest
studied elevation range taxa spring
(m a.s.l.); found average
microhabitat (min–
considered max)

Amacuzac 3 Rh-ca; 800 100 21 62 17 47 – Aphanothece microscopica, Chamaecalyx swirenkoi, Valadez-

River Blennothrix ganeshii, Calothrix Hapalosiphon Cruz
Basin braunii, Chamaesiphon welwitschii, et al.
confervicola, Microcoleus Stichosiphon (1996)
Biodivers Conserv (2015) 24:865–888

lacustris, Phormidium retzii, sansibaricus,

Pleurocapsa minor, Scytonema Xenococcus bicudoi,
coactile Xenococcus lamellosus

C. coccoid; F. filamentous non-heterocytous; H. heterocytous; %Cy. % of cyanobacteria out of all benthic algae; He helocrenes; Li limnocrenes; Rh rheocrenes; Hy
hygropetric springs; si siliceous substratum; ca carbonate substratum; SAL spring-associated-limestone sources; ISP inland saline springs; Ir iron springs; Su sulfur springs;
RG rock–glacier springs; F fountains. Microhabitat considered: El rocky material (epilithon), Eb bryophytes (epibryon), Ss surface sediment, Ep bryophytes ? vascular plants
(epiphyton). Nomenclature of older papers was updated
Author's personal copy
875

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 Table 2 Cyanobacteria in ambient springs: environmental determinants and adaptations
Environmental Adaptations of
determinant spring cyanobacteria
123
High irradiance and Photoprotection. Selective
high UV adsorption of UV radiation
Shading Chromatic adaptation
‘‘Purple’’ species
Discharge fluctuations,
desiccation
Resting stages
Nutrients: nitrogen Diazotrophism
Nutrients: phosphorus Utilization of organic
phosphorus compounds
Chemotaxis
Strong current Highly-effective adhesion
mechanisms, tough and
flexible filaments
Structure or Taxa
pigment involved
Scytonemin, MAAs – Cantonati et al. (2012b, 2014a);
Phycoerithrin, phycocyanin, Chamaesiphon starmachii, C.
chlorophyll. Variable ratios confervicola, Pleurocapsa
aurantiaca
As above but stable ratios. Heteroleibleinia purpurascens,
Phormidium tinctorium
Thick sheaths Calothrix parietina, Chamaesiphon
polonicus, C. starmachii,
Pleurocapsa aurantiaca,
Gloeocapsa spp.
Heterocytes Tolypothrix penicillata
Heterocytes in SAL springs to fix Myochrotes myochrous, Petalonema
atmospheric N during alatum
desiccation periods
Hairs – Cantonati et al. (2012b); Muñoz-
Calyptra – Muñoz-Martı́n et al. (2014)
Adhesion pads Chamaesiphon geitleri, C.
starmachii, C. fuscus, Clastidium
rivulare, C. setigerum,
Pleurocopsa aurantiaca,
Stichosiphon pseudopolymorphus,
Tapinothrix varians,
Heteroleibleinia purpurascens
876
Reference
Castenholz and Garcia-Pichel
(2012)
Cantonati et al. (2012b, 2014b)
Cantonati et al. (2012b, 2014b)
Cantonati (2008); Cantonati et al.
(1996, 2012b); Maier (1995); Rott
et al. (1999); Gesierich and Kofler
(2010); Gesierich and Rott (2004)
Cantonati (2008); Cantonati et al.
(1996, 2012a, b, c); Gesierich and
Rott (2004)
Gesierich and Kofler (2010)
Author's personal copy
Martı́n et al. (2014)
Cantonati (2008); Cantonati et al.
(1996, 2012b); Gesierich and
Kofler (2010); Backhaus (2006);
Gesierich and Rott (2004)
Biodivers Conserv (2015) 24:865–888
 Table 2 continued

Environmental Adaptations of Structure or Taxa Reference

determinant spring cyanobacteria pigment involved

?Grazing, Toxin (microcystin) – Rivularia spp., Tolypothrix distorta, Aboal et al. (2005)
?competition with production Schizothrix fasciculata
other
photoautotrophs
‘‘?’’ indicates that the exact adptive meaning of ofcyanotoxin production in springs still has to be elucidated
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could be pointed out (siliceous/carbonate): Chamaesiphon fuscus/C. geitleri, Tapinothrix

janthina (Fig. 1c)/T. varians (Cantonati et al. 1996, 2012b).
Flowing springs on siliceous substratum often host rich cyanobacterial assemblages
including taxa adapted to low conductivity and alkalinity (Chamaesiphon starmachii,
Fig. 1b; Tapinothrix janthina, Fig. 1c), high discharge and strong current (Chamaesiphon
fuscus; Heteroleibleinia purpurascens, Fig. 1d) (Table 2) (Cantonati et al. 2012b).

Rheocrenic springs on carbonate substratum

The few attempts to characterize spring types by means of benthic algae including
cyanobacteria usually point out different kinds of flowing springs on carbonate substratum
e.g.: mid-altitude, oligotrophic rheocrenes; and low-altitude, mostly shaded, slightly
N-enriched rheocrenes. The former are characterized by desiccation-tolerant (Pleurocapsa
aurantiaca), oligotraphentic (Xenotholos kerneri, Fig. 1k), and rheophilic (Tapinothrix
varians) cyanoprokaryotes, whilst the latter host eutraphentic (Pleurocapsa minor; Phor-
midium retzii, Fig. 1f), and large-discharge-preferring (Chamaesiphon geitleri) species
(Cantonati et al. 2012b).
Phormidium retzii was found by Dell’Uomo (1990) to be the dominant cyanoprokaryote
species in a large, low-elevation (100 m a.s.l.), slow-flowing rheocrene (Su Gologone karst
spring) draining limestones on the Isle of Sardinia.

Hygropetric springs

Rock-face seepages are more common on carbonate substratum. Cantonati et al. (1996)
found the highest number of cyanobacterial taxa in a carbonate hygropetric spring, and

Fig. 2 Some representative, characteristic, and rare cyanoprokaryotes from warm-climate ambient-temperature
springs. a Symplocastrum muelleri, b Blennothrix ganneshi, c Hapalosiphon welwitschii, d Chamaecalyx
swirenkoi, e Homoeothrix juliana, f Stichosiphon exiguus, g Xenotholos huastecanus (scale bar = 50 lm),
h Chamaesiphon confervicola (scale bar = 10 lm), i Homoeothrix juliana, j Stichosiphon sansibaricus,
k Hyella kalligrammos, l Merismopedia cf. glauca. If not otherwise stated, the scale bars equal 5 lm

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attributed this finding to the fact that hygropetric springs include transition microhabitats to
other biotopes (dripping and wet rock walls). In agreement with this result, Gesierich and
Kofler (2010) noted a large proportion of pseudaerial taxa (e.g., from the genus Gloeo-
capsa) in a carbonate rock-face seepage (Table 2). Rivularia spp. (Figure 1h), Plectonema
tomasinianum, Ammatoidea normannii, Chamaesiphon minutus (mainly as an epiphyte of
other cyanobacteria), and Calothrix parietina were the cyanobacterial indicator taxa of this
spring type identified by Cantonati et al. (2012b).

Spring-associated-limestone (SAL) sources

SAL springs possess very distinctive benthic-algae assemblages in which cyanoprokaryotes

are dominant: Phormidium incrustatum, Tapinothrix crustacea, Myochrotes myochrous
(Fig. 1i), Dichothrix gypsophila, Gloeothece confluens, and Gloeocapsopsis spp. (Cantonati
et al. 2012b). Gesierich and Kofler (2010) found a large SAL spring to be dominated by large
black cushions of an association of pseudaerial cyanobacteria: Myochrotes myochrous and
Petalonema alatum. The relevance of Phormidium incrustatum in SAL springs was con-
firmed by Pentecost (2003), who stated that the species has a worldwide distribution (span-
ning latitudes 54°N–30°S), and that it is common on shaded spring-deposited travertines.

Inland saline springs

According to Garcı́a and Aboal (2014), Aboal and Garcı́a (personal com.), Cantoral-Uriza
and Aboal-Sanjurjo (2010), the characteristic cyanobacterial taxa of saline springs within a
marsh close to the sea and in an arid climate setting were: Calothrix pulvinata, Phormi-
dium tergestinum, and Tapinothrix violacea (Table 1). MC (unpublished data) found an
inland saline spring draining evaporite formations to include a species of Rivularia (R. sp.
aff. bullata) among the main macroalgae.

Iron springs

Iron springs are often acidic and dominated by filamentous green algae and iron bacteria
(Cantonati et al. 2012b). Accordingly, no cyanoprokaryotes were found by Gesierich and
Kofler (2010) in an iron spring dominated by green algae. Cantonati (2008) found only a
larger thallus of Phormidium corium in an iron spring but this colony thrived on leaf litter
only and did not develop on the stones, heavily encrusted by iron and manganese pre-
cipitates. Guasch et al. (2012) found no cyanobacterial denaturing gradient gel elec-
trophoresis (DGGE) operational taxonomic units (OTUs) in the spring mouth and in the
uppermost part of a canal fed by an iron spring.

Cyanobacteria in ambient springs in warm climate settings (tropical, subtropical,

arid) (Fig. 2a–l)

Garcı́a and Aboal (2014), Aboal and Garcı́a (personal com.), Cantoral-Uriza and Aboal-San-
jurjo (2010) found the following cyanobacterial taxa to be indicators of freshwater carbonate
spring conditions in a coastal marsh: Calothrix parietina, Homoeothrix juliana (Fig. 2e, i),
Leptolyngbya angustissima, Schizothrix fasciculata, Tapinothrix crustacea (Table 1).

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In 14 wells (bir) and four ambient-temperature springs (ain) in the El-Farafra Oasis in
the Western Desert of Egypt, mostly affected by direct human and cattle impact (organic
pollution), the main cyanoprokaryote taxa were the following: Oscillatoria jasorvensis var.
thermalis, Jaaginema thermale, Homeothrix juliana, Oscillatoria animalis, O. princeps, O.
limosa (Saber 2015; personal communication).
Ambient springs in warm climates host a rich and diverse cyanobacterial com-
munity, still poorly-known so far (Table 1). More in-depth, calcareous warm springs
(23–30 °C) are characterized by a complex community that includes epilithic, epi-
phytic, and metaphytic forms. In North America, Whitford (1956) studied 30 springs
in Florida State (USA), with temperatures from 21 to 30 °C. The springs studied
included, hard-water freshwater, oligohaline, mesohaline, and sulphide springs. He
reported 35 species of cyanobacteria. In hard-water freshwater springs the cyanobac-
teria found were: Amphithrix sp., Calothrix sp., Lyngbya epiphytica, Lyngbya kutzingii,
Lyngbya nordgardhii, Oscillatoria splendida, Lyngbya wollei (as Plectonema wollei),
and Xenococcus sp.
Recently Wehr (personal com.) found in Volusia Blue Spring, a karstic spring in Florida
State, Chroococcus sp., Heteroleibleinia kuetzingii, several unidentified species of Lep-
tolyngbya and Oscillatoria, including O. cf. limosa, Phormidium cf. retzii, Pleurocapsa cf.
minor; and the rare Stichosiphon willei, and Xenotholos kerneri.
Several calcareous springs have been studied in the central region of Mexico. Carmona-
Jiménez and Montejano (1993) and Montejano et al. (2000) studied several karstic springs,
with temperatures ranging from 22 to 37 °C (Table 1). Blennothrix ganneshi (Fig. 2b),
Homoeothrix juliana, and Symplocastrum cf. muelleri were widely distributed in all
springs. Particularly noteworthy is the epiphytic cyanobacterial community that includes
common species, such as Chamaesiphon confervicola (Fig. 2h), Chlorogloea epiphytica,
Chamaechalyx swirenkoi (Fig. 2d), Cyanocystis mexicana, Stichosiphon filamentosus,
Stichosiphon sansibaricus (Fig. 2j), Xenococcus bicudoi, Xenococcus willei, and Xeno-
tholos huastecanus (Fig. 2g). Rare species include Stichosiphon exiguus (Fig. 2f) and
Chamaecalyx calyculatus. In habitats with low light intensity, as springs emerging in
caves, it is common to find a special community, dominated by endolithic species of Hyella
(Fig. 2k), that grows together with the red alga Hildenbrandia angolensis.
Valadez-Cruz et al. (1996) studied a calcareous warm spring in the Hidalgo State, in
central México, with temperatures ranging from 29 to 30 °C. Blennothrix ganeshii was the
most conspicuous cyanoprokaryote. Also common were Hapalosiphon welwitschii
(Fig. 2c), Microcoleus lacustris, Chamaesiphon confervicola, Phormidium retzii, and
Scytonema coactile. The epiphytic community includes Chamaecalyx swirenkoi, Sti-
chosiphon sansibaricus, Xenococcus bicudoi, and Xenococcus lamellosus.
A special situation was described by Garcia-Pichel et al. (2002) in a bottom-fed artesian
spring in the Mexican Chihuahuan Desert (Cuatro Cienegas karstic region). Cyanobacterial
centimetre-sized waterwarts, formed by an Aphanothece-like cyanobacterium, where
suspended within a central well by upwelling waters, and supported a community of
epiphytic filamentous cyanobacteria. Waterwarts contained calcite crystals, likely needed
as ballast to prevent washing out of the well.
In a small spring in São Paulo State (southeastern Brazil) with contrasting (very-low)
conductivity, two of the four occurring macroalgae were found to be cyanobacteria:
Lyngbya putealis, and Scytonema arcangeli (Necchi 1992).

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Cyanobacterial adaptations to the spring habitat

Ambient-temperature springs are characterized by a large variety of environmental con-

ditions, requiring a diverse set of adaptations to the cyanobacteria that colonize them
(Table 2).
As regards light exposures, adaptation to radiation range from UV exposure to deep
shade, and are similar to those observed for benthic cyanoprokaryotes in lakes with fluc-
tuating water level (Cantonati et al. 2014a, b). Cyanobacteria in high-elevation, hy-
gropetric, SAL springs thus often have yellow-brownish sheaths because of the presence of
the UV absorbing compound scytonemin (Castenholz and Garcia-Pichel 2012). By con-
trast, cyanoprokaryotes in forest springs are often purple-violet, because of permanently or
temporarily (chromatic adaptations) increased phycoerithrin-Chl a and/or phycocyanin-Chl
a ratios.
As regards water-level fluctuations, it might be recalled that many springs, especially on
carbonate substratum, are fed by small and surficial aquifers, and are thus highly unstable.
Cyanoprokaryotes thriving in them thus need to possess adaptations to withstand exposure
(see above) and desiccation. The latter consist usually in mucilages and thick sheaths: a
typical example is Chamaesiphon polonicus, a widespread taxon very common also in
springs both on carbonate and siliceous substratum.
As regards nutrient availability, nitrogen limitation is very rare, even in remote
mountain locations due to diffuse airborne pollution (Waldner et al. 2014). This might
explain the low proportion of heterocytous taxa often found in springs (Table 1). An
exception is represented by SAL springs, in which heterocytous cyanobacteria are well
represented. Gesierich and Kofler (2010) hypothesized that the high proportion of hete-
rocytous taxa in a SAL spring with largely developed hygropetric microhabitat might be an
adaptive strategy to warrant nitrogen supply in periods of desiccation. Diazotrophism in
springs is however likely to be much more widespread than can be suggested by the
occurrence and abundance of heterocytes. On the contrary, phosphorus is commonly
limiting in mountain and oligotrophic springs (e.g., Cantonati et al. 2012c). Adaptations by
cyanoprokaryotes colonizing springs include the production of phosphatases to gain access
to P in organic compounds. This is especially well documented for species provided with
so-called ‘‘hairs’’ (filamentous forms with long tapering ends). Another adaptation of lothic
cyanobacteria to P scarcity is the presence of calyptras, that are supposed to be involved in
the chemotactic location of P-rich microzones (e.g., Muñoz-Martı́n et al. 2014). Phormi-
dium favosum (‘‘autumnale’’ group), one of the most widespread Phormidia in spring
habitats, commonly possesses calyptras on many apices in springs (Fig. 1e).
As regards water motion, large rheocrenes on siliceous substratum and karstic springs
can have important discharge and seasonally strong currents. Cyanobacteria thus need to
firmly anchor themselves to the substratum with adhesion pads etc. The most common
spring cyanoprokaryote species include a large proportion of rheophilic taxa (Table 2).
Aboal et al. (2005) demonstrated that cyanobacterial toxin production is widespread
also in high-ecological-integrity calcareous Mediterranean streams. Since several of the
species for which microcystins production was shown (Rivularia spp., Tolypothrix dis-
torta, Schizothrix fasciculata) occur also in springs, it is very likely that toxins might be
produced by cyanobacteria in ambient springs as well (Table 2). The exact adaptive
meaning of cyanotoxin production in high-integrity running waters still has to be elu-
cidated. One of the most plausible interpretations is that cyanoprokaryotes may use these
toxins for successful competition for space and nutrients with other primary producers, in
particular other algae, and as an anti-grazing defence. This might be particularly

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important in resource-poor habitats and mostly of small to very-small dimensions, such

as springs.

Impacts and threats for ambient-spring cyanobacteria conservation

In spite of the many impacts affecting spring habitats, springs and spring organisms,
including cyanobacteria, are not at all or little considered by conservation legislation in the
different countries (Cantonati et al. 2012a). The main threat for spring-habitats remains
their systematic use for drinking water supply. This implies the destruction of the original
morphology, a severe impoverishment of the biota (e.g., Zollhöfer 1997), and, in particular,
the disappearance of photoautotrophs. Spring capturing and tapping to obtain drinking
water or to generate hydro-electric power will increase even more in the future as a result
of climate change, with a reduction of precipitation being predicted for many areas.
Springs are further exposed to a series of direct (e.g., forest management) and indirect (e.g.,
nutrient or contaminant deposition in the drainage basin) impacts. Because of the small
dimensions and of the importance of the fringing semi-aquatic habitats, springs are ex-
tremely sensitive to disturbance factors, such as trampling damage by cattle, coverage by
sediment, stripping off of the surrounding vegetation, nutrient inputs etc. (e.g., Gesierich
and Kofler 2010, Niedermayr and Schagerl 2010).
Springs in near-natural conditions are continuously reduced in numbers even in pro-
tected areas. Strong and organised initiatives for spring conservation have been undertaken
only in a few countries (cf. Hotzy 2007). Springs are not especially mentioned within the
WFD (European Water Framework Directive, EU-WFD 2000), although the use of phy-
tobenthos (in the restricted sense of diatoms plus macrophytes, rarely all algae including
cyanobacteria and diatoms) is recommended as one key element, beside macroinverte-
brates, for the assessment of ecological water quality in rivers. There is only a general
comment given that attention should be deserved to the interfaces between groundwater
and surface waters.
Only a very-limited number of spring types is mentioned in the Annex I (‘‘Natural
habitat types of community interest whose conservation requires the designation of special
areas of conservation’’) of the 1992 European Union Directive on the conservation of
natural habitats and of wild fauna and flora:—SAL springs, ‘‘7220 Petrifying springs with
tufa formation (Cratoneurion)’’, priority habitat type;—some salt springs, ‘‘1340 Inland
salt meadows (Puccinellietalia distantis)’’;—some mineral-rich springs, ‘‘7160
Fennoscandian mineral-rich springs and spring-fens’’ (EU-HD 1992; Evans 2006; Jokic
and Galz 2007).
Springs and groundwater use is in general regulated by specific legislation, mostly
oriented toward drinking water use related prevention of microbiological contamination.
These regulations require the excavation of the spring mouth down to the bedrock (or
aquifer), and the construction of closed, mostly concrete, housing including several de-
position basins (i.e. a complete spring capturing). This kind of protection is exclusively
oriented toward the use of spring water for drinking water supply but ignores completely
the value of springs as natural habitats. In most disagreements over multiple uses of water,
at the international or local scale, assignment of water to maintain aquatic biodiversity is
usually disregarded (Poff et al. 2003). It is well known that springs are a fundamental
source of good quality water and they frequently occupy a place of distinction in cultures
and mythologies. On the contrary there is still very limited public awareness on the fact
that they are also special habitats of great importance for nature conservation. Biodiversity

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conservation is a critical test of whether water exploitation or freshwater-habitat alterations

are sustainable. This assumption is a foundation for the use of freshwater organisms as
biomonitors of habitat quality and integrity (Dudgeon et al. 2006).
Freshwater habitats are biodiversity rich but unluckily also human-activities’ hotspots.
This has led to widespread habitat degradation (Strayer and Dudgeon 2010). The threats to
global freshwater biodiversity can be grouped under five interacting categories (Dudgeon
et al. 2006): (1) flow alteration, (2) habitat destruction or degradation, (3) over-exploita-
tion, (4) pollution, and (5) invasion by non-native species (e.g., Allan and Flecker 1993).
Global-scale environmental changes, such as nitrogen deposition (e.g., Waldner et al.
2014), warming, and modifications in patterns of precipitation are superimposed upon all
these threat categories. An emblematic example relative to ambient springs, cyanobacteria,
and allochthonous and invasive species is provided by Florida springs. Stevenson et al.
(2007) showed that almost all large Florida springs had macroscopic algae growing in
them, an average of 50 % of the spring bottoms were covered by macroalgae, and
thickness of macroalgal mats was commonly half a meter. A cyanobacterium, Lyngbya
wollei, was one of the two most common invasive taxa (together with the xanthophyte
Vaucheria spp.).
For freshwater springs in the area of the Pego-Oliva marsh (East Iberian Peninsula) for
which long-term data were available, a change in the distribution and abundance of
Schizothrix fasciculata was observed. While 30 years ago it was more abundant in the
freshwater springs studied, it reduced its occurrence in these habitats but increased its
abundance in the source of a river that was also under observation. This species was found
to prefer low nutrient concentrations, and the changes in its abundance might be a response
to the increasing anthropogenic influences on the marsh (Cantoral-Uriza and Aboal-San-
jurjo 2010; Garcı́a-Fernández 2014; Garcı́a and Aboal 2014).
Garcı́a-Fernández (2014), and Garcı́a and Aboal (2014), while studying the macroalgae
of the Pego-Oliva marsh (see above) found that the springs, the less disturbed habitats of
the marsh, were rich in taxa linked to oligotrophic conditions and that have restricted
distributions. These species can be considered a LIHRe sensu Cantonati et al. (2012a).
Among drainage basins and water bodies there is relevant species diversification at
smaller geographic scales, and many freshwater species have restricted distribution (e.g.,
Strayer et al. 2004). These features combine with endemism to generate a ‘‘lack of sub-
stitutability’’ among freshwater habitats. It follows that protection of one or a few fresh-
water habitats cannot preserve all biodiversity in a region, or even a substantial proportion
of it (Dudgeon et al. 2006). Investigating springs in the Dolomiti Bellunesi National Park,
about 50 % of the cyanoprokaryotes and algae other than diatoms found could reasonably
be considered rare on the basis of the literature and experience (Cantonati 2008). The high
total number of taxa and the low diversity of the individual sites (Table 1) highlighted
again the marked heterogeneity of spring habitats, and the importance of protecting large
number of springs to preserve aquatic biodiversity (Nascimbene et al. 2011).
In addition, all the main microhabitats should be considered in biodiversity inventories,
as relevant differences among substrata have been demonstrated (Table 1; Cantonati et al.
2012a).
Best management practices (BMP) that are meanwhile established for streams, such as
restoration of old structures no longer in use or more sustainable ways of spring capturing,
are only very seldom applied to springs (and only to very specific spring types, in particular
SAL springs; Jokic and Galz 2007).
Cantonati et al. (2009) had to face the tapping for water abstraction by the local
municipality of one of their long-term monitoring springs (Gerecke et al. 2011). This

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allowed them to show the potential value of residual habitats for spring biodiversity
conservation. They also noted the paramount importance of substrata, and of their exact
location, in determining which cyanobacteria taxa will colonize the residual habitat.

Conclusions and perspectives

Due to both under-appreciation of spring studies, and increasingly complex (and expen-
sive) approaches required for the characterization of cyanobacteria assemblages in near-
natural habitats, the literature on cyanoprokaryotes in ambient-temperature springs is very
limited.
However, some generalizations are possible. The marked heterogeneity of the spring
habitat requires a diverse set of adaptations (Table 2). Different types of ambient springs
are thus colonized by rheophilic, UV and radiation resistant, shade-tolerant, non-
diazotrophic, P-specialist (hairs and calyptras) cyanoprokaryotes. Cyanobacteria can suc-
cessfully be used to statistically characterize the diverse types of ambient springs (Can-
tonati et al. 2012b). Some morphotypes, e.g., Phormidium retzii in large karstic springs at
low elevation, Homeothrix juliana in ambient springs in arid regions, are regularly reported
from these habitats even from distant geographic locations. Coccoid types appear to
dominate in ambient springs in temperate climate while filamentous types appear to prevail
in ambient springs in warm and arid climates (Table 1).
The in-depth characterization of ambient-spring cyanoprokaryote assemblages is im-
portant because springs are biodiversity-rich but highly menaced ecotones. Moreover, if
adequately known, spring cyanoprokaryotes might provide a valid contribution to the
assessment of the ecological integrity, quality, and value for nature protection of spring
habitats.
If traditional morphotaxonomy is limited by its inability to unveil cryptic diversity, that
appears to characterize many cyanobacterial taxa (Dvořák et al. 2014, 2015), purely
metagenomic approaches to natural communities (e.g., DGGE, and cloning of the 16 S
rRNA gene) are often scarcely informative on the nature and ecological attributes of the
OTUs found (e.g., Junier et al. 2013), and even next generation sequencing approaches are
limited by the availability and correctness of data in molecular databases (GenBank etc.).
Thus, morphological, microscopic analysis still remains the quickest comprehensive
approach to the assessment of cyanobacterial diversity in ambient springs (compare
Manoylov 2014). However, given that these special habitats are still largely understudied,
the likelihood to find also problematic, rare, and new types is high. Molecular and phy-
logenetic studies are a valid support in these studies. These types are however often
difficult and/or time-consuming to cultivate. Therefore, the improvement of isolation and
single-cell and filament sequencing methods and techniques (Mareš et al. 2015) will be
very important for research on cyanoprokaryotes in spring habitats. This will be relevant
also in the light of the fact that some of the most widespread taxa in ambient springs (e.g.,
Phormidium retzii), are known to hide cryptic species with different genotypes (Casamatta
et al. 2003). Improved molecular tools could thus allow to recognize spring-specific taxa
out of these apparently ‘‘cosmopolitan’’ species. In general, more widespread use of
molecular markers in the study of cyanobacteria in crenic habitats might help overcome
problems related to the fact that the descriptive literature on cyanobacterial taxonomy is
sometimes controversial, and that morphospecies identification can be doubtful when
morphological variability is high.

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Most studies of ambient-springs cyanobacteria focused on the epilithon. Other substrata

of the spring microhabitat mosaic need to be more in-depth explored. In particular the
application of recent methods developed for the sampling of lake epipelon (Yang and
Flower 2009; Yang et al. 2010; Poulı́čková et al. 2014) might greatly enhance our
knowledge on cyanobacteria in helocrenic springs.
Quantitative studies (e.g., Necchi 1992; Spitale et al. 2012b) occur still relatively rarely.
The same is true for functional aspects. Exceptions are provided by in-depth studies on the
ecophysiology of Lyngbya wollei from Florida springs (e.g., Sickman et al. 2009), and by
the investigation by Camacho et al. (2005), who showed how different ecophysiological
strategies, such as resistance and/or use of oxygen and sulfide, light adaptation, or resis-
tance to high current, allow each of the different microorganisms, including
cyanoprokaryotes, to efficiently colonize several areas within the environmental gradient a
cold sulfur spring. Functional aspects of ambient-spring cyanobacteria deserve more at-
tention in the future.
Some topics that urgently need to be developed in spring research in general (Cantonati
et al. 2012a) should be studied also with a focus on cyanoprokaryotes: continued devel-
opment of habitat-type and geographic-area specific indices to assess the quality of spring
habitats, development and testing of new strategies for non-destructive spring capturing
and for restoration of impacted habitats, improvement of long-term ecological research.

Acknowledgments MC was partially funded by the Autonomous Province of Trento while contributing to
this study. We are grateful to Dr. Nicola Angeli (Museo delle Scienze—MUSE, Limnology and Phycology
Research Unit, Trento, Italy) for the layout of Fig. 1.

References

Aboal M, Puig MÁ, Asencio AD (2005) Production of microcystins in calcareous Mediterranean streams:
the Alharabe River, Segura River basin in south-east Spain. J Appl Phycol 17:231–243
Allan JD, Flecker AS (1993) Biodiversity conservation in running waters. Bioscience 43:32–43
Backhaus D (2006) Litorale Aufwuchsalgen im Hoch- und Oberrhein. Carolinea 64:5–68
Camacho A, Rochera C, Silvestre JJ, Vicente E, Hahn M (2005) Spatial dominance and inorganic carbon
assimilation by conspicuous autotrophic biofilms in a chemical gradient of a cold sulfurous spring: the
role of differential ecological strategies. Microb Ecol 50:172–184
Cantonati M (2008) Cyanoprokaryotes and algae other than diatoms in springs and streams of the Dolomiti
Bellunesi National Park (Northern Italy). Algol Stud 126:113–136
Cantonati M, Rott E, Pipp E (1996) Ecology of cyanophythes in mountain springs of the River Sarca
catchment (Adamello-Brenta Regional Park, Trentino, Northern Italy). Algol Stud 83:145–162
Cantonati M, Bertuzzi E, Scalfi A, Campana V (2009) The potential importance for spring conservation of
residual habitats after flow capturing: a case study. Verh Int Ver Limnol 30:1267–1270
Cantonati M, Angeli N, Bertuzzi E, Spitale D, Lange-Bertalot H (2012a) Diatoms in springs of the Alps:
spring types, environmental determinants, and substratum. Freshw Sci 31:499–524
Cantonati M, Füreder L, Gerecke R, Jüttner I, Cox EJ (2012b) Crenic habitats, hotspots for freshwater
biodiversity conservation: toward an understanding of their ecology. Freshw Sci 31:463–480
Cantonati M, Rott E, Spitale D, Angeli N, Komárek J (2012c) Are benthic algae related to spring types?
Freshw Sci 31:481–498
Cantonati M, Guella G, Spitale D, Angeli N, Borsato A, Lencioni V, Filippi ML (2014a) The contribution of
lake benthic algae to the sediment record in a carbonate mountain lake influenced by marked natural
water-level fluctuations. Freshw Sci 33:499–512
Cantonati M, Komárek J, Hernández-Mariné M, Angeli N (2014b) New and poorly-known coccoid species
(Cyanoprokaryota) from the mid-depth and deep epilithon of a carbonate mountain lake. Freshw Sci
33:548–556
Cantoral-Uriza EA, Aboal-Sanjurjo M (2010) Benthic algal communities of some sources of the Oliva-Pego
marsh, Valencia, Spain. Limnetica 29:287–300 (in Spanish with English summary)

123
 Author's personal copy
886 Biodivers Conserv (2015) 24:865–888

Carmona-Jiménez J, Montejano G (1993) Caracterización ficológica de manantiales de la cuenca baja del

sistema hidrológico del Pánuco, México. Bol Soc Bot México 53:21–41
Casamatta DA, Vis ML, Sheath RG (2003) Cryptic species in cyanobacterial systematics: a case study of
Phormidium retzii (Oscillatoriales) using RAP molecular markers and 16S rDNA sequence data. Aquat
Bot 77:295–309
Castenholz RW (1969) Thermophilic blue–green algae and the thermal environment. Bacteriol Rev
33:476–504
Castenholz RW, Garcia-Pichel F (2012) Cyanobacterial responses to UV radiation. In: Whitton BA (ed) The
ecology of cyanobacteria II: their diversity in space and time. Springer, Dordrecht, pp 481–499
Dell’Uomo A (1975) La flora e le associazioni algali della Sorgente di Salomone (Anterselva, Bolzano).
Facies estiva. Giorn Bot Ital 109:257–271
Dell’Uomo A (1990) Chorological analysis of the algal flora of ‘‘Su Gologone’’ spring in Sardinia. Inter-
national Symposium on: biogeographical aspects of insularity (Rome, 18–22 May 1987). Accad. Naz.
Lince, Rome. Atti Convegni Lincei 85:417–427
Dudgeon D, Arthington AH, Gessner MO, Kawabata Z, Knowler D et al (2006) Freshwater biodiversity:
importance, threats, status, and conservation challenges. Biol Rev 81:163–182
Dvořák P, Casamatta DA, Poulı́čková A, Hašler P, Ondřej V, Sanges R (2014) Synechococcus: 3 billion
years of global dominance. Mol Ecol 23:5538–5551
Dvořák P, Poulı́čková A, Hašler P, Belli M, Casamatta DA, Papini A (2015) Species concepts and speciation
factors in cyanobacteria, with connection to the problems of diversity and classification. Biodivers
Conserv (in press)
EU-HD (European Union Habitat Directive) (1992) Council directive 92/43/EEC of 21 May 1992 on the
conservation of natural habitats and of wild fauna and flora (EC habitats directive). Off J Eur Com L
206:7–50
EU-WFD (European Union Water Framework Directive) (2000) Directive 2000/60/EC of the European
Parliament and of the Council of 23. October 2000 establishing a framework for Community action in
the field of water policy. Off J Eur Com L 327:1–72
Evans D (2006) The habitats of the European Union Habitats Directive. Biol Env Proc R Irish Acad
106:167–173
Garcı́a ME, Aboal M (2014) Environmental gradients and macroalgae in Mediterranean marshes: the case of
Pego-Oliva marsh (East Iberian Peninsula). Sci Tot Environ 475:216–224
Garcı́a-Fernández ME (2014) Biodiversity of continental macroalgae and ecological quality assessment in
transitional waters. The case of Pego-Oliva marsh (E Iberian Peninsula). PhD thesis. Universidad de
Murcia, Facultad De Biologı́a, Murcia, Spain
Garcia-Pichel F, Wade BD, Farmer JD (2002) Jet-suspended, calcite-ballasted cyanobacterial waterwarts in
a desert spring. J Phycol 38:420–428
Gerecke R, Cantonati M, Spitale D, Stur E, Wiedenbrug S (2011) The challenges of long-term ecological
research in springs in the northern and southern Alps: indicator groups, habitat diversity, and medium
term change. J Limnol 70(Suppl. 1):168–187
Gesierich D, Rott E (2004) Benthic algae and mosses from aquatic habitats in the catchment of a glacial
stream (Rotmoos, Ötztal, Austria). Ber nat-med Ver Innsbruck 91:7–42
Gesierich D, Kofler W (2010) Are algal communities from near-natural rheocrene springs in the Eastern
Alps (Vorarlberg, Austria) useful ecological indicators? Algol Stud 133:1–28
Glazier DS (2009) Springs. In: Likens GE (ed) Encyclopedia of inland waters, vol 1. Academic, San Diego,
pp 734–755
Golubić S (1967) Algenvegetation der Felsen. In: Thienemann A (ed) Die Binnengewässer 23. Sch-
weizerbart’s, Stuttgart
Guasch H, Garcia-Acosta X, Urrea G, Bañeras L (2012) Changes in the microbial communities along the
environmental gradient created by a small Fe spring. Freshw Sci 31:599–609
Hájek M, Roleček J, Cottenie K, Kintrová K, Horsák M, Pouličková A, Hájková P, Franková M, Dı́tě D
(2011) Environmental and spatial controls of biotic assemblages in a discrete semi-terrestrial habitat:
comparison of organisms with different dispersal abilities sampled in the same plots. J Biogeogr
38:1683–1693
Hájková P, Bojková J, Franková M, Opravilova V, Hájek M, Kintrová K, Horsák M (2011) Disentangling
the effects of water chemistry and substratum structure on moss dwelling unicellular and multicellular
micro-organisms in spring-fens. J Limnol 70:54–64
Hašler P, Poulı́čková A (2005) Cyanobacteria of the spring fens of a part of West Carpathians. Biologia
60:1–7

123
 Author's personal copy
Biodivers Conserv (2015) 24:865–888 887

Hotzy R (2007) Spring protection in Bavaria. In: Cantonati M, Bertuzzi E, Spitale D (eds) The spring
habitat: biota and sampling methods, vol 4., Monografie del Museo Tridentino di Scienze Natural-
iMuseo Tridentino di Scienze Naturali, Trento, pp 317–326
Hu B-F, Xie S-L (2013) Determination of effects of environmental factors on stream macroalgae in Xin’ an
spring by grey correlation analysis. Bull Bot Res 33:98–103
Jokic Z, Galz S (2007) Life-Natur-Projekt. Kalktuffquellen in der Frankenalb. Tagungsband (Proceedings).
Landesbund für Vogelschutz in Bayern e.V., Verband für Arten- und Biotopschutz (LBV). Hilpolt-
stein, Germany
Junier P, Kim O-S, Imhoff JF, Witzel K-P, Hadas O (2013) Effect of salinity on cyanobacterial community
composition along a transect from Fuliya spring into the water of Lake Kinneret, Israel. Fund Appl
Limnol 182:99–107
Kann E (1978) Systematik und Ökologie der Algen österreichischer Bergbäche. Arch Hydrobiol 53:405–643
(suppl)
Komárek J, Kann E (1973) Zur Taxonomie und Ökologie der Gattung Homoeothrix. Arch Protistenk
115:173–233
Komárek J, Golubić S (1990) Guide to the nomenclature and formal taxonomic treatment of oxytroph
prokaryotes (Cyanoprokaryotes). Proposal, p 114. Třeboň and Boston
Maier M (1995) Der Algenaufwuchs in zwei geologisch unterschiedlichen Fliessgewässern (Litz und Meng)
in Vorarlberg unter besonderer Berücksichtigung jahreszeitlicher Veränderungen. Algol Stud
77:95–109
Manoylov KM (2014) Taxonomic identification of algae (morphological and molecular): species concepts,
methodologies, and their implications for ecological bioassessment. J Phycol 50:409–424
Mareš J, Lara Y, Dadáková I, Hauer T, Uher B, Wilmotte A, Kaštovský J (2015) Phylogenetic analysis of
the cultivation-resistent genera Stigonema and Petalonema (Nostocales, Cyanobacteria) using single-
cell and filament sequencing of environmental samples. J Phycol. doi:10.1111/jpy.12273
Montejano G, Carmona Jiménez J, Cantoral-Uriza E (2000) Algal communities from calcareous springs and
streams in La Huasteca, central Mexico: a synthesis. In: Munawar M, Lawrence SG, Munawar IF,
Malley DF (eds) Aquatic ecosystems of Mexico: status and scope. Backhuys, Leiden, The Netherlands,
pp 135–149
Muñoz-Martı́n MÁ, Martı́nez-Rosell A, Perona E, Fernández-Piñas F, Mateo P (2014) Monitoring
bioavailable phosphorus in lotic systems: a polyphasic approach based on cyanobacteria. Sci Tot
Environ 475:158–168
Nascimbene J, Spitale D, Thüs H, Cantonati M (2011) Congruencies between photoautotrophic groups in
springs of the Italian Alps: implications for conservation strategies. In: Cantonati M, Gerecke R,
Jüttner I, Cox EJ (eds) Springs: neglected key habitats for biodiversity conservation. J Limnol 70(suppl
1):3–8
Necchi O (1992) Macroalgae dynamics in a spring in São Paulo State, southeastern Brazil. Arch Hydrobiol
124:489–499
Niedermayr R, Schagerl M (2010) Structuring factors of the phytobenthos community along a mountain
headwater (National Park Kalkalpen, Austria). Fund Appl Limnol 177:93–104
Pentecost A (2003) Taxonomic identity, ecology and distribution of the calcite-depositing cyanobacterium
Phormidium incrustatum (Oscillatoriaceae). Cryptog Algol 24:307–321
Poff NL, Allan JD, Palmer MA, Hart DD, Richter BD et al (2003) River flows and water wars: emerging
science for environmental decision-making. Front Ecol Environ 1:298–306
Poulı́čková A, Dvořák P, Mazalová P, Hašler P (2014) Epipelic microphototrophs: an overlooked assem-
blage of lake ecosystems. Freshw Sci 33:513–523
Rott E, Pfister P, Pipp E, Pall K, Binder N, Ortler K (1999) Indikationslisten für Aufwuchsalgen in
Fließgewässern Österreichs. Teil 2: Trophieindikation und autökologischen Anmerkungen. WWK,
Bundesministerium Land- und Forstwirtschaft, Wien (ISBN 3–85 174–25–4), p 248
Saber AA (2015) Algae of El-Farafra oasis. PhD thesis. Faculty of Science, Ain Shams University, Egypt
Sanders D, Wertl W, Rott E (2010) Spring-associated limestones of the Eastern Alps: overview of facies,
deposystems, minerals, and biota. Facies (online first). doi:10.1007/s10347-010-0252-y
Sciuto K, Moro I (2015) Cyanobacteria: the bright and dark side of a charming group. Biodivers Conserv (in
press)
Sickman JO, Albertin A, Anderson MW, Pinowska A, Stevenson RJ (2009) A comparison of internal and
external supply of nutrients to algal mats in two first magnitude springs in Florida. J Aquat Plant
Manag 47:135–144
Spitale D, Leira M, Angeli N, Cantonati M (2012a) Environmental classification of springs of the Italian
Alps and its consistency across multiple taxonomic groups. Freshw Sci 31:563–574

123
 Author's personal copy
888 Biodivers Conserv (2015) 24:865–888

Spitale D, Lencioni V, Cantonati M (2012b) Relative importance of space and time in determining the biotic
structure in the upper part of spring-fed streams. Freshw Sci 31:586–598
Starmach K (1929) Über polnische Chamaesiphon Arten. Acta Soc Bot Pol 6:30–45
Stevenson RJ, Pinowska A, Albertin A, Sickman JO (2007) Ecological condition of algae and nutrients in
Florida springs: the synthesis report. Florida Department of Environmental Protection, Tallahassee
Strayer DL, Dudgeon D (2010) Freshwater biodiversity conservation: recent progress and future challenges.
J North Am Benth Soc 29:344–358
Strayer D, Downing JA, Haag WR, King TL, Layer JB et al (2004) Changing perspectives on pearly
mussels, North America’s most imperiled animals. Bioscience 54:429–439
Uher B, Skácelová O, Kováčik L (2001) Sinice několika studánek v okolı́ Brna (Cyanobacteria of several
wells in Brno surroundings) (in Czech). Czech Phycol 1:21–30
Valadez-Cruz F, Carmona-Jiménez J, Cantoral-Uriza E (1996) Algas de ambientes lóticos del Estado de
Morelos, México. Anales Ins Biol Univ Nac Autón México Ser Bot 67:227–282
Vis ML, Sheath RG (1996) Distribution and systematics of Batrachospermum (Batrachospermales,
Rhodophyta) in North America. 9. Section Batrachospermum: descriptions of five new species. Phy-
cologia 35:124–134
Waldner P, Marchetto A, Thimonier A, Schmitt M, Rogora M et al (2014) Detection of temporal trends in
atmospheric deposition of inorganic nitrogen and sulphate to forests in Europe. Atmos Environ
95:363–374
Ward DM, Castenholz RW, Miller SR (2012) Cyanobacteria in geothermal habitats. In: Whitton BA (ed)
Ecology of cyanobacteria II: their diversity in space and time. Springer, Dordrecht, pp 39–63
Whitford LA (1956) The communities of algae in the springs and spring streams of Florida. Ecology
37:433–442
Whitford LA, Schumacher GJ (1963) Communities of algae in North Carolina streams and their seasonal
relations. Hydrobiologia 22:133–196
Yang H, Flower RJ (2009) A portable hand-operated sampler for shallow water surface sediments with
special reference to epipelic communities. J Paleolimnol 42:317–324
Yang H, Flower RJ, Battarbee RW (2010) An improved coverslip method for analysing epipelic diatom. Eur
J Phycol 45:191–199
Zollhöfer JM (1997) Quellen die unbekannten Biotope: erfassen, bewerten, schützen. Bristol-Schriftenreihe
6:1–153, Bristol-Stiftung, Zürich

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