Extremophiles (2016) 20:603–620
DOI 10.1007/s00792-016-0849-3
ORIGINAL PAPER
Diversity of extremophilic bacteria in the sediment
of high‑altitude lakes located in the mountain
desert of Ojos del Salado volcano, Dry‑Andes
Júlia Margit Aszalós1 · Gergely Krett1 · Dóra Anda1 · Károly Márialigeti1 ·
Balázs Nagy2 · Andrea K. Borsodi1 
Received: 15 March 2016 / Accepted: 31 May 2016 / Published online: 17 June 2016
© Springer Japan 2016
Abstract  Ojos del Salado, the highest volcano on Earth is
surrounded by a special mountain desert with extreme arid-
ity, great daily temperature range, intense solar radiation,
and permafrost from 5000 meters above sea level. Several
saline lakes and permafrost derived high-altitude lakes can
be found in this area, often surrounded by fumaroles and
hot springs. The aim of this study was to gain informa-
tion about the bacterial communities inhabiting the sedi-
ment of high-altitude lakes of the Ojos del Salado region
located between 3770 and 6500 m. Altogether 11 sediment
samples from 4 different altitudes were examined with 16S
rRNA gene based denaturing gradient gel electrophoresis
and clone libraries. Members of 17 phyla or candidate divi-
sions were detected with the dominance of Proteobacteria,
Acidobacteria, Actinobacteria and Bacteroidetes. The bac-
terial community composition was determined mainly by
the altitude of the sampling sites; nevertheless, the extreme
aridity and the active volcanism had a strong influence on
it. Most of the sequences showed the highest relation to
bacterial species or uncultured clones from similar extreme
environments.
Keywords  High-altitude lakes · Dry-Andes · Bacterial
diversity · 16S rRNA gene · DGGE · Clone library
Communicated by A. Oren.
* Andrea K. Borsodi
borsodi.andrea@ttk.elte.hu
1
Department of Microbiology, Eötvös Loránd University,
Pázmány Péter sétány 1/C, 1117 Budapest, Hungary
2
Department of Physical Geography, Eötvös Loránd
University, Pázmány P. sétány 1/C, 1117 Budapest, Hungary
Introduction
Ojos del Salado (6893 m, S 27°06′34.6″, W 68°32′32.1″),
the highest volcano on Earth is a dormant stratovolcano
hosted by the deserted plateau of Puna de Atacama (with
an average altitude of 4500 meters above sea level; m.a.s.l)
in the Dry-Andes. As an effect of the South American Dry
Diagonal, the plateau is characterized by extreme arid-
ity (lack of glaciers and increased elevation of potential
snowline altitude), forming a remote mountain desert up to
6000 m (Vuille and Ammann 1997; Ammann et al. 2001;
Houston and Hartley 2003; Azócar and Brenning 2010;
Nagy et al. 2014a).
Mountain deserts are harsh environments character-
ized by extreme aridity, intense solar UV radiation and
extreme shifts in daily temperature (even 50 °C on the
surface). In the region of Ojos del Salado precipitation
occurs sporadically, and the snow rapidly sublimates
instead of melting (Nagy et al. 2014a). In this hyper-
arid environment, altitudinal limit of vascular plants and
continuous vegetation is 4600 m.a.s.l., much lower than
in the case of the more humid Himalaya (6350 m.a.s.l.)
(Halloy 1991).
In mountain deserts water is present mainly in the form
of ice within permafrost. Permafrost by definition of the
Permafrost Subcommittee (1988) is “ground (i.e., soil and
rock) that remains at or below 0 °C for at least 2 years”.
In the region of Ojos del Salado from 5000 m.a.s.l discon-
tinuous, while from 5600 m.a.s.l continuous permafrost can
be found. The active layer of permafrost thaws periodically
as a result of temperature changes. Although, the melting
affects only the upper 5–60 cm of the permafrost, the active
layer slowly increases as an effect of global climate change
(Nagy et al. 2014b). Meltwater from thawing permafrost
frequently accumulates in endorheic basins forming small
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and shallow high-altitude lakes. These permafrost origi-
nated lakes are among the highest altitude lakes on Earth.
Nevertheless, in a longer period of time they might disap-
pear due to the degradation of permafrost and desiccation
of the regolith (Nagy et al. 2014a, b).
Beside lakes derived from the degrading permafrost,
several salt-flats and high-altitude saline lakes (e.g. Laguna
Santa Rosa, Laguna Verde) can also be found in the Puna
de Atacama. The volumes of these high-altitude Dry-
Andean lakes are decreasing since the last humid period
because evaporation outweighs the amount of precipitation
(Demergasso et al. 2010).
The high-altitude lakes of Ojos del Salado are extreme
environments resulting from the properties of mountain
deserts, including intense solar radiation, radical changes in
daily temperature and low water activity. Volcanic activities
also have an influence on the physical and chemical char-
acteristics of the environment; several hot springs are pre-
sent in this area which can alter the pH, and—along with
the solar radiation—affect the water temperature in a wide
range. Fumaroles in high-altitude sites have been reported
to host diverse communities with mosses, lichens, fungi
and microbial mats in otherwise blank places (Halloy 1991;
Costello et al. 2009).
High-altitude lakes are sensitive ecosystems which
respond rapidly to environmental changes; therefore, they
are natural laboratories and indicators for climate changes
(Adrian et al. 2009). They also give us insights into early
life on Earth when the lack of ozone layer caused intense
UV radiation and extremities in temperature (Cabrol et al.
2007). These lakes might be inhabited by several hitherto
unknown microorganisms with special adaptive strategies
to the extreme environmental conditions (Ordoñez et al.
2009). Former studies focused mainly on other lakes of the
Dry-Andes (Demergasso et al. 2008; Dorador et al. 2008,
2013; Cabrol et al. 2009; Farías et al. 2009, 2014; Lynch
Fig. 1  Geographic location of Ojos del Salado (red star) in the map of South-America (a) and the sampling sites (yellow crosses) (b)
13
Extremophiles (2016) 20:603–620
et al. 2012; Scott et al. 2015), and research was also per-
formed on similar environments, for example Antarctic
lakes and permafrost (Goordial et al. 2016; Aislabie et al.
2013; Antibus et al. 2012; Cowan et al. 2002) and high-alti-
tude lakes or permafrost of the Tibetan Plateau (Wu et al.
2006; Xing et al. 2009; Zhang et al. 2007). Understanding
the microbial processes in degrading permafrost is also an
important issue, since thawing is proven to cause shifts in
microbial communities, functional gene abundances which
might be responsible for changes in global carbon cycle
(Mackelprang et al. 2011; Tas et al. 2014; Hultman et al.
2015).
The aim of the present work was to reveal the hitherto
unknown bacterial communities inhabiting the sediments
of five different high-altitude lakes located in the Ojos del
Salado volcano, Dry-Andes. To gain information about the
bacterial diversity and compare the community structures,
16S rRNA gene based denaturing gradient gel electropho-
resis and molecular cloning methods were applied.
Materials and methods
Description of sampling sites
The studied five high-altitude lakes are located near the
border between Chile and Argentina in the area of Ojos del
Salado volcano. Geographical location and GPS coordinates
of the lakes are indicated in Fig. 1 and Table 1. Laguna Santa
Rosa and Laguna Verde are permanent saline lakes located at
3770 and 4350 m.a.s.l., respectively. Because of the intense
evaporation (1000 mm year−1) and very small amount of
precipitation (170 mm year−1), the surface area of Laguna
Santa Rosa and Laguna Verde has been constantly shrinking
(Hiner 2009). This phenomenon caused an increase in the
salt concentration of the lake water and accumulation of salt
Extremophiles (2016) 20:603–620 605

Table 1  Geographic parameters of the lakes in the area of Ojos del Salado volcano, location and physical characteristics of the sampling sites,
numerical data of DGGE and 16S rRNA gene clone library analysis
Laguna
Laguna Santa Rosa Lake I Lake II Lake III
Verde
Elevation (m.a.s.l.) 3770 4350 5900 5900 6500
Lake area (km2, estim.) 2 15 0.0025 0.0025 0.006
Maximum depth (m, estim.) 1.2 4 1 1 1

Sample code AS-12 AS-13 AS-14 AS-5 AS-6 AS-9 AS-10 AS-1 AS-2 AS-3 AS-4
27.081085 S 27.081085 S 26.890288 S 27.088495 S 27.088495 S 27.088360 S 27.088396 S 27.109925 S 27.110914 S 27.110900 S 27.110206 S
GPS coordinates
69.175022 69.175022 68.486858 68.534330 68.534330 68.531410 68.531532 68.550184 68.550108 68.550121 68.550885
Water temperature (°C) 7 7 36 7 7 7 7 0 36 36 2
Air temperature (°C) 5 5 7 0 0 0 0 -5 -5 -5 -5
pH 9.5 9.5 8.8 6.6 6.6 6.8 6.8 4.8 2.1 2.1 4.8
Conductivity (μS cm-1) 6540 6540 6260 1230 1230 1081 1542 n.e. 1360 n.e. n.e.
Sediment sampling depth (cm) 0-5 10-15 0-5 0-5 10-15 20 0-5 0-5 0-5 0-5 0-5
Number of DGGE bands 14 30 22 27 32 26 26 13 5 9 15
Number of ARDRA groups n.e. 43 42 27 n.e. n.e. 42 28 25 n.e. n.e.
Number of identified molecular clones n.e. 38 34 21 n.e. n.e. 33 27 24 n.e. n.e.
Clones with >97% similarity to a described species n.e. 4 1 7 n.e. n.e. 19 1 0 n.e. n.e.
Clones with >97% similarity to a molecular clone n.e. 13 2 7 n.e. n.e. 4 9 7 n.e. n.e.

ne not examined, estim. estimated

crystals on the lakeside. Laguna Verde is fed by several cold
and warm (34–36 °C) springs. Two seasonal nameless lakes
(Lake I and II) found at 5900 m.a.s.l., are derived from the
melting permafrost, and both can be characterized by shal-
low clear water bodies. These lakes were partially ice cov-
ered at the time of sampling. The third seasonal lake (Lake
III) is located at 6500 m.a.s.l. This lake is surrounded by sev-
eral fumaroles and fed by meltwater of perennial snowfields
(1 °C) and a small warm (36 °C) creek. At the time of sam-
pling this lake was completely frozen down to the bottom.
Presence of warm springs near Laguna Verde and the warm
spring and fumaroles at the 6500 m elevation site are signs of
volcanic activity.
Sampling was accomplished during the Földgömb Ata-
cama Climate Monitoring Expedition in February 2014.
Samples were collected into 50 ml sterile Falcon tubes
from different sediment layers of all five lakes as indicated
in Table 1, and were stored between −5 and 10 °C until
being processed in the laboratory.
DNA extraction and PCR amplification
The community DNA was isolated using the Ultra Clean
Soil Kit (MO Bio Inc., CA, USA) according to the manu-
facturer’s instructions. The 16S rRNA gene was amplified
by PCR using Bacteria-specific 27 forward (5′-AGAGTTT
GATCMTGGCTCAG-3′) (Lane 1991) and 1401 reverse
(5′-CGGTGTGTACAAGACCC-3′) (Nübel et al. 1996)
primers. The following temperature protocol was used
for bacterial PCR: initial denaturation at 95 °C for 5 min,
followed by 32 cycles of denaturation at 94 °C for 30 s,
annealing at 52 °C for 45 s and elongation at 72 °C for
60 s, and a final extension at 72 °C for 10 min. The PCR
reaction mixture contained 200 µM of each deoxynu-
cleoside triphosphate, 1 U of LC Taq DNA Polymerase
(recombinant) (Fermentas, Lithuania), 1X Taq buffer with
(NH4)2SO4 (Fermentas, Lithuania), 2 mM MgCl2, 0.65 mM
of each primer, and approximately 20 ng of genomic DNA
template in a total volume of 25 µL. Both community DNA
isolates and PCR products were visualized in 1 % agarose
gel stained with ECO Safe Nucleic Acid Staining Solution
(Avegene, Taiwan) using UV excitation.
Denaturing gradient gel electrophoresis (DGGE)
A semi-nested PCR was carried out on the 16S rRNA gene
amplicons using a GC-clamp (5′-GC clamp-CAGAGTTT-
GATCCTGGCTCAG-3′) containing 27 forward and 519
reverse (5′-GTNTTACNGCGGCKGCTG-3′) primers
(Muyzer et al. 1993). The reaction mixture and temperature
protocol were the same as mentioned above. Electrophore-
sis of PCR products was run for 14 h using an INGENY
phorU-2 electrophoresis system in a 7 % polyacrylamide
gel containing 40–60 % gradient of denaturants (40 %
formamide and 7 M urea is defined as 100 %) in 1X TAE
buffer at 100 V and 60 °C. Following the run, the gel was
stained with ethidium-bromide, and detected under UV
light.
Discrete bands were excised from the gel with a sterile
scalpel, and after an overnight incubation in 30 μl of dis-
tilled water, the extracted DNA was amplified with the 27
forward and 519 reverse primers using the same reaction
mixture and temperature profile as described previously.
The DGGE profiles of the samples were compared, and
an UPGMA (unweighted pair group method using arithme-
tic mean algorithm) dendrogram (Smalla et al. 2007) was
constructed based on the position and presence of bands in
the different lanes (representing the samples) using Total
Lab (TL 120) version 2006 (Nonlinear Dynamics Inc.,
Newcastle upon Tyne, UK) software.

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606
Construction of molecular clone libraries
The purified (EZ-10 Spin Column PCR Purification Kit,
Bio Basic, Canada) 27 forward—1401 reverse PCR prod-
ucts were ligated into a TA-cloning vector (pGEM-T Vec-
tor System, Promega, WI, USA) according to the manu-
facturer’s instructions, and transformed into competent E.
coli JM109 cells. For blue/white selection, the transformed
cells were spread on LB agar plates containing 100 µg ml−1
ampicillin, 80 µg ml−1 X-Gal and 0.5 mM IPTG, and incu-
bated overnight at 37 °C. White colonies were picked for
the clone libraries, and DNA was extracted from the E. coli
cells by incubating the cultures at 98 °C for 5 min, and pel-
leting the cell fragments by centrifugation with 4500 rcf for
5 min. Insert sequences of each clone were amplified with
standard M13 forward (5′-GTAAAACGACGGCCAGT-3′)
and M13 reverse (5′-CAGGAAACAGCTATG-3′) primers
(Messing 1983) followed by a nested PCR with the original
27 forward and 1401 reverse primers. The thermal profiles
of PCRs were the same as described previously.
ARDRA grouping and rarefaction analysis
PCR amplicons were grouped on the basis of their ampli-
fied ribosomal DNA restriction analysis (ARDRA) patterns
produced with restriction enzymes as described by Massol-
Deya et al. (1995). The applied restriction enzymes (Hin6I
and BsuRI, Fermentas, Lithuania) were selected based on
their different specificity (3′-G^CGC-5′ and 3′-GG^CC-5′,
respectively) that resulted in different restriction pattern of
PCR products allowing us to make ARDRA groups (Krett
et al. 2013; Borsodi et al. 2012). Each reaction mixture
contained 2 µl R/Tango Buffer (Fermentas), 7.76 µl ster-
ile distilled water, 0.24 µl of enzyme and 10 µl of PCR
product. Digestions were made at 37 °C, for 3 h. Diges-
tion products were separated in 2 % agarose gel on 80 V
for 80 min, stained with ECO Safe Nucleic Acid Staining
Solution (Avegene, Taiwan), and visualized by UV exci-
tation. Clones were grouped according to their different
ARDRA patterns. At least one representative from each
ARDRA group was sequenced.
Rarefaction analysis was carried out to check, whether
the number of clones analyzed was satisfactory to estimate
diversity within the clone libraries and to compare the diver-
sity of clone libraries. Rarefaction curves were produced
with the software Past3 (Hammer et al. 2001) by plotting
the number of 16S rRNA gene clones in each clone libraries
against the number of clones in different ARDRA groups.
Sequencing, identification and phylogenetic analysis
PCR products from the excised DGGE bands and representa-
tive clones of each ARDRA group were sequenced with 27
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Extremophiles (2016) 20:603–620
forward primers using the automated Sanger-method by LGC
Ltd. (Berlin, Germany). The quality of each chromatogram
was checked with the help of the Chromas software, and low
quality ends were trimmed (Technelysium Pty Ltd., Aus-
tralia). Taxonomic relationships of the partial 16S rRNA gene
sequences were determined by EzTaxon-e database (Kim et al.
2012). The 16S rRNA gene sequences were deposited into
GenBank under accession numbers LN929548-LN929732.
In the case of molecular clones, phylogenetic relations
of sequences at least 97 % similarity to an uncultured envi-
ronmental clone or described species (Stackebrandt and
Goebel 1994; Vandamme et al. 1996; Tindall et al. 2010)
found in GenBank were analyzed by construction of neigh-
bor joining trees with MEGA6 software (Tamura et al.
2013). Sequences (along with the nearest cultivated rela-
tive or molecular clone sequences derived from GenBank)
were aligned by ClustalW (Larkin et al. 2007), the number
of bootstrap replications was 1000 and Kimura 2-parameter
(Kimura 1980) model was applied.
Results and discussion
Denaturing gradient gel electrophoresis
To compare the bacterial community structures of the sedi-
ment samples from the lakes located in the Ojos del Salado
region, similarity dendrogram was constructed on the basis
of the DGGE patterns (Fig. 2). Three groups were formed
according to the altitude of the studied lakes. Samples from
the highest altitude were clearly separated from the others;
furthermore, they formed pairs (AS-2 and AS-3; AS-1 and
AS-4) in relation to the different temperatures of the sampling
sites. Samples from Lake I (AS-5, AS-6) and Lake II (AS-
9, AS-10) located at 5900 m.a.s.l. formed the second group.
The two sediment samples (AS-12 and AS-13) from Laguna
Santa Rosa (3770 m.a.s.l.) formed the third group together
with the sample (AS-14) from Laguna Verde (4350 m.a.s.l.).
It is interesting to note that although sample AS-14 derived
from a warm environment, it did not contain typical common
bands with the other two warm samples (AS-2 and AS-3).
Altogether 85 different bands were detected from the gel.
Number of DGGE bands for each sample is indicated in
Table 1. The smallest amount of bands (thus the least diverse
community) was revealed in samples derived from 6500 m.a.s.l.
(AS-1, AS-2, AS-3, AS-4), especially in the sediments of the
warm shallow creek (AS-2, AS-3). Number of bands from sam-
ples at 3770 and 5900 m.a.s.l. consisted of 14–32 bands without
a clear relationship to the altitude of the lakes or the depth of
sediment. At the same time, samples from lakes I and II found at
5900 m.a.s.l. contained surprisingly large amount of bands (26–
32) suggesting high genetic diversity. In addition, the difference
between the band numbers of samples AS-12 (30) and AS-13
Extremophiles (2016) 20:603–620 607

Fig. 2  Similarity UPGMA
dendrogram of the DGGE
molecular fingerprints of the
sediment samples from lakes
of Ojos del Salado (elevation is
indicated after the code of the
sample. The excised bands are
marked with arrows)

Table 2  Phylogenetic affiliation of 16S rRNA gene sequences of the excised DGGE bands according to EzTaxon
DGGE band Sample code Accession number Similarity (%) E value Coverage (%) Closest taxon in EzTaxon (accession number)

Firmicutes
 An_9 AS-12 LN929727 97 (179/184) 7.00E−75 100 Halanaerobium sehlinense (JN381500)
Gemmatimonadetes
 An_3 AS-6 LN929726 85 (384/450) 8.00E−113 100 Gemmatimonas aurantiaca (AP009153 )
Proteobacteria
 An_29 AS-1 LN929732 98 (461/469) 0 100 Rhodanobacter umsongensis (FJ821731)
 An_1 AS-4 LN929724 91 (418/461) 2.00E−169 100 Thiobacillus thiophilus (EU685841)
 An_14 AS-13 LN929731 98 (461/471) 0 100 Thiobacillus thiophilus (EU685841)
 An_2 AS-6 LN929725 98 (429/436) 0 100 Noviherbaspirillum psychrotolerans (JN390675)
 An_13 AS-9 LN929730 99 (463/470) 0 94 Noviherbaspirillum psychrotolerans (JN390675)
 An_10 AS-10 LN929728 97 (398/412) 4.00E−175 100 Brevundimonas faecalis (FR775448)
 An_11 AS-10 LN929729 98 (460/471) 0 100 Thermomonas brevis (AJ519989)

(14) was more than twice, despite the fact that both originated
from Laguna Santa Rosa located at 3770 m.a.s.l.
From the DGGE gel a total of 14 discrete bands were
excised and sequenced out of which 9 were successfully
identified. They belonged to three different phyla (Firmi-
cutes, Gemmatimonadetes and Proteobacteria) as it is indi-
cated in Table 2. At species level, DGGE sequences were
closely related to e.g., the extreme halophilic Halanaerobium
sehlinense, firstly isolated from the Tunisian hypersaline lake
Sehline Sebkha (Abdeljabbar et al. 2013) and the psychro-
tolerant Noviherbaspirillum psychrotolerans, described from
Larsemann Hills, Antarctica (Bajerski et al. 2013).
16S rRNA gene based molecular clone libraries
Based on the DGGE similarity dendrogram, 6 sediment
samples (at least one from each altitude) were chosen for
a detailed phylogenetic analysis by molecular clone librar-
ies. Both samples AS-1 and AS-2 were derived from the
upper 0–5 cm sediment layer at the highest altitude sam-
pling site (Lake III), from a cold (0 °C) and a warm (36 °C)
environment, respectively. From the lakes at 5900 m.a.s.l.
two samples were chosen. Both samples AS-5 (Lake I) and
AS-10 (Lake II) were taken from 0 to 5 cm sediment depth
but on the DGGE similarity dendrogram they clearly sepa-
rated from each other. Clone libraries from Laguna Verde
(sample AS-14) and Laguna Santa Rosa (sample AS-13)
were also examined in detail.
Following the ARDRA grouping, 177 representatives
were sequenced and identified from the altogether 431
molecular clones. The rarefaction curves (Fig. 3) did not
reach asymptotes, indicating that further diversity would
have been revealed by the analysis of a higher number of
clones. The presence of the most diverse communities was
found in the saline lakes at lower altitudes (samples AS-13,

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608 Extremophiles (2016) 20:603–620

AS-14) while the highest elevation sites (samples AS-1,

AS-2) hosted the least diverse bacterial assemblages on the
basis of rarefaction curves.
In the six samples, members of 17 different phyla or
candidate divisions (Aquificae, Deinococcus-Thermus,
Chloroflexi, Nitrospirae, Cyanobacteria, Chlorobi, Proteo-
bacteria, Firmicutes, Actinobacteria, Acidobacteria, Bacte-
roidetes, Verrucomicrobia, Caldithrix, Gemmatimonadetes,
TM7, OD1, WS5) were detected. However, major differ-
ences were observed in the composition of the bacterial
assemblages from different sampling sites. The relative
abundance of molecular clones among the bacterial phyla
per sample is shown in Fig. 4.
Members of Proteobacteria were present in each clone
library, and in 4 cases (AS-1, AS-2, AS-5, AS-13) Betapro-
teobacteria was the most abundant. The AS-10 clone library
Fig. 3  Rarefaction curves for the different ARDRA patterns of 16S
was predominated with Gammaproteobacteria. In AS-14, rRNA gene clones from lake sediment samples of the lakes located in
unlike other clone libraries, the class Betaproteobacteria the area Ojos del Salado volcano
was not detected but the members of Epsilonproteobacteria
were present. Representatives of Acidobacteria, Bacteroi-
detes, Actinobacteria and Cyanobacteria were also detected
at least in three clone libraries.
The clone library constructed from the sediment sam-
ple of the extreme saline Laguna Santa Rosa (AS-13) con-
tained 10 phyla, and showed the most diverse community
structure amongst the examined lakes. Proteobacteria was
the most abundant phylum with four classes (Alpha-, Beta-,
Gamma-, and Deltaproteobacteria), but members of Aci-
dobacteria, Bacteroidetes, Actinobacteria, Chloroflexi and
Verrucomicrobia were also abundant (Fig. 4). In this clone
library altogether 37 ARDRA groups were separated. How-
ever, most of the molecular clones showed less than 97 %
similarity to any described species. Only a few molecular
clones (Fig. 5) could be identified at species level (with
higher than 97 % of 16S rRNA gene sequence similarity
to described species) (Stackebrandt and Goebel 1994; Van-
damme et al. 1996; Tindall et al. 2010). They were closely
related to the species Rhodoferax ferrireducens (Betapro-
teobacteria), the first described non-phototrophic member
of the genus (Finneran et al. 2003) which grows via dis-
similatory Fe(III) reduction, and R.  antarcticus (Madigan Fig. 4  Distribution of members of different phyla in the 16S rRNA
et al. 2000), a moderately psychrophilic (growth optimum gene clone libraries constructed from sediment samples of the lakes
of Ojos del Salado, Dry-Andes
15–18 °C) purple non-sulfur bacterium first isolated from
Antarctic microbial mats. Representatives of the species
Gillisia hiemivivida (Bowman and Nichols 2005) were pre- that the largest ARDRA group of this sample showed the
sent in this clone library, too. It is a psychrophilic and halo- closest similarity to an uncultured environmental clone
philic bacterium. Sequences closely related to the chemo- (Par-s-7, EF632909) from the high-altitude Chilean Alti-
lithotrophic nitrite-oxidizing species of Nitrospira marina plano (Dorador et al. unpublished data).
(Watson et al. 1986) was also detected. Sequences showing
96 % similarity to species Herminiimonas glaciei (Love- Fig. 5  Neighbor-joining phylogenetic tree based on the 16S rRNA ▸
gene sequence data of bacterial clones from sediment samples of
land-Curtze et al. 2009) detected in this clone library rep-
the lakes of Ojos del Salado showing >97 % similarity to described
resented one of the dominant DGGE bands in the sample species. The number of members of the ARDRA groups is indicated
from Lake II found at 5900 m.a.s.l. It is worth mentioning after the representative clone

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Extremophiles (2016) 20:603–620 609

AS-10-79 (LN929646) / 2 clones

96
79 AS-5-51 (LN929610) / 2 clones
AS-10-47 (LN929635) / 1 clone
55 71 Caenimonas terrae strain SGM1-15 (GU181268) / Soil (Korea)

99 AS-10-55 (LN929638) / 7 clones

AS-5-3 (LN929598) / 2 clones
45 98
Acidovorax delafieldii (AF078764) / n.d.
94 Acidovorax facilis strain THWCSN37 (GQ284428) / Natural spring sediment (India)
63 A S-13-23 (LN929661) / 2 clones
70
AS-13-3 (LN929653) / 3 clones Betaproteobacteria
100
Rhodoferax ferrireducens (AF435948) / Subsurface sediment, Oyster Bay (USA)
100
AS-5-79 (LN929615) / 1 clone
100 Polaromonas cryoconiti strain Cr4-35 (HM583567) / Pitztaler Jöchl glacier cryoconite (Austria)

AS-10-59 (LN929640) / 1 clone

100 99 Hydrogenophaga palleronii (AF019073) / n.d.
86 AS-10-76 (LN929645) / 1 clone
99
AS-5-20 (LN929602) / 3 clones
63 AS-10-30 (LN929628) / 1 clone
100
Janthinobacterium svalbardensis strain JA-1 (DQ355146) / Austre Broggerbreen glacier (Spitzbergen)
100 AS-1-71 (LN929568) / 1 clone
Rhodanobacter umsongensis strain GR24-2 (FJ821731) / Soil of a ginseng field (China)
100 AS-10-83 (LN929647) / 3 clones
99
93 Pseudoxanthomonas wuyuanensis (JN247803) / Saline-alkali soil (China) Gammaproteobacteria
99 AS-10-16 (LN929625) / 7 clones
93
Thermomonas brevis GZ436 (KF923805) / Soil of a coal mine (China)
71 AS-10-34 (LN929631) / 2 clones
95 Lysobacter koreensis (AB166878) / Soil of a ginseng field (Korea)
61
AS-10-95 (LN929651) / 2 clones Gammaproteobacteria
100 Cellvibrio fibrivorans R4079 (AJ289164) / Soil (Belgium)
100 AS-10-6 (LN929620) / 4 clones
Brevundimonas variabilis strain ATCC 15255 (AJ227783) / n.d.
100 AS-10-4 (LN929619) / 1 clone
68 100
Devosia glacialis strain Cr4-44 (HM474794) / Pitztaler Jöchl glacier cryoconite (Austria) Alphaproteobacteria
100 AS-10-42 (LN929633) / 4 clones
62
Catellibacterium aquatile strain A1-9 (EU313813) / Daqing reservoir freshwater (China)
100 AS-14-7 (LN929695) / 1 clone
41 100 Roseinatronobacter monicus strain ROS 35 (DQ659236) / Mono Lake (USA)
AS-13-82 (LN929687) / 1 clone Nitrospira
100 Nitrospira marina (X82559) / Heating system (Russia)
100 AS-10-31 (LN929629) / 10 clones
Microcoleus vaginatus PBP-D-KK1 (EF654072) / n.d. Cyanobacteria
100 AS-10-60 (LN929641) / 5 clones
100 Crinalium epipsammum SAG 22.89 (AB115964) / n.d.
100 AS-10-48 (LN929636) / 1 clone
Hymenobacter roseosalivarius strain AA718 (Y18833) / Soil (Antarctica)
99 AS-5-56 (LN929612) / 1 clone

100
100
AS-10-9 (LN929622) / 1 clone
Pedobacter daechungensis strain Dae 13 (AB267722) / Soil of a ginseng field (China)
100 AS-13-18 (LN929658) / 2 clones Bacteroidetes
94
Gillisia hiemivivida strain IC154 (AY694006) / Maritime habitat (Antarctica)
99 AS-5-29 (LN929604) / 10 clones
100
Flavobacterium glaciei strain 0499 (DQ515962) / Glacier (China)
100 AS-5-80 (LN929616) / 7 clones
92 Flavobacterium psychrophilum strain IFO 15942 (AB078060) / n.d.
100 AS-10-10 (LN929623) / 1 clone
Actinobacteria
Nocardioides alpinus strain Cr7-14 (GU784866) / Alpine glacier cryoconite (Austria)
78 AS-10-11 (LN929624) / 1 clone Deinococcus-Thermus
100 Deinococcus marmoris strain AA-63 (JNIV01000230) / Marble rock (Antarctica)

0.05

AS-1: 6500 m (cold water)

AS-2: 6500 m (warm water)
AS-5: 5900 m (cold water)
AS-10: 5900 m (cold water)
AS-14: 4350 m (warm water)
AS-13: 3770 m (cold water)

13

610
The clone library constructed from the sediment sample
of the extreme saline Laguna Verde (AS-14) was unique
among the studied samples. Members of altogether 6 phyla
were present in this clone library (Fig. 4) including four
classes of Proteobacteria (Alpha, Gamma- Delta- and Epsi-
lonproteobacteria). Most of the identified molecular clones
showed less than 97 % similarity to any environmental
clones or formerly described species of bacteria (Figs. 5,
6). The most abundant groups showed only 90–91 % simi-
larity to environmental clones from the phyla Caldithrix
and Chloroflexi, respectively. Only one minor representa-
tive could be identified as member of species Roseina-
tronobacter monicus (Boldareva et al. 2007), an alkaliphi-
lic halotolerant Alphaproteobacteria described from the
hypersaline Mono Lake, California. Other ARDRA group
representatives showed the highest similarity to uncul-
tured clones from hot environment of methane-rich mud
volcanoes, anaerobic hydrothermal vents or hot-springs in
Greenland, Columbia or the Philippines. The relatively low
sequence similarity rates suggest high abundance of unde-
scribed species within this sample.
Bacterial communities hosted by two permafrost derived
lakes found at 5900 m.a.s.l. were very similar according to
the clone libraries AS-5 and AS-10 (Fig. 4). The two clone
libraries consisted of members of 7 phyla and shared many
similar sequences, and both of them included molecular
clones closely related to ones described from cryoconites,
glaciers, permafrost sediments or freshwater lakes (Fig. 6).
Among the molecular clone representatives, species
Janthinobacterium svalbardensis (Avguštin et al. 2013)
was detected in both samples. This psychrotrophic (growth
occurs at 2–25 °C) bacterium was first described from a
glacier in the Spitsbergen. Members of the genus Flavo-
bacterium were also present in both clone libraries, and
formed the most abundant ARDRA group in sample AS-5.
Representatives of cold-adapted species F. glaciei (Zhang
et al. 2006) and F. psychrophilum (Nakagawa et al. 2002)
were detected in the sediment of Lake I (AS-5). Simi-
larly, species F. psychrolimnae (Van Trappen et al. 2005)
was described from microbial mats in the Antarctic Lake
Fryxell, representatives of which were exposed from sedi-
ment sample AS-10 of Lake II. Betaproteobacteria was the
second most abundant group in AS-5, and the presence of
species Polaromonas cryoconitii was detected. This psy-
chrophilic bacterium was first isolated from a cryoconite of
Pasterze/Großglockner glacier in the Hohe Tauern, Austria
(Margesin et al. 2012).
Interestingly, in sample AS-10 members of phylum
Cyanobacteria were also detected, and according to the
13
Extremophiles (2016) 20:603–620
Fig. 6  Neighbor-joining phylogenetic tree based on the 16S rRNA ▸
gene sequence data of bacterial clones from sediment samples of the
lakes of Ojos del Salado showing >97 % similarity to an uncultured
bacterial clone. The number of members of the ARDRA groups is
indicated after the representative clone
number of molecular clones they were dominant, however,
this phylum was absent in sample AS-5 (Fig. 4).
A minor ARDRA group in sample AS-10 was closely
related to Hymenobacter roseosalivarius, a species of
phylum Bacteroidetes (Fig. 5). This strictly aerobic, oli-
gotrophic bacterium was isolated from soil samples of the
Antarctic polar desert, Dry Valleys (Hirsch et al. 1998).
In sample AS-10, only one molecular clone was related
to Actinobacteria. It showed 97 % similarity to species
Nocardioides alpinus, an actinomycete isolated from a
cryoconite of Pitztaler Jöchl glacier in the Ötztaler Alps in
Tyrol, Austria (Zhang et al. 2012).
AS-10 was the only sample where presence of Chlorobi
was observed (Fig. 4). A molecular clone showed high sim-
ilarity to an unknown environmental sequence from moss
pillars in a freshwater lake, Hotoke Ike, Antarctica (Nakai
et al. 2012).
The only member of the phylum Deinococcus-Thermus
in this study was detected in the sample AS-5 (Fig. 4).
Deinococcus marmoris is, a desiccation and UV radiation
tolerating bacterium was described from Antarctic marble
(Hirsch et al. 2004). Molecular clones related to the mem-
bers of Gemmatimonadetes were also present in the sample
AS-5 (Fig. 4). These clones showed the highest similarity
to other environmental sequences, for example an environ-
mental clone derived from Mendenhall glacier, USA (Sat-
tin et al. 2009) and another from Socompa volcano in the
Dry-Andes (Costello et al. 2009).
Two sediment samples (AS-1 and AS-2) from Lake
III found at 6500 m.a.s.l derived from relative proximity,
however, have different physical and chemical characteris-
tics (Table 1). AS-1 was a sediment sample from the shore
affected by melt water of the ice body of the lake. AS-2
was a sediment sample under the warm and extremely
acidic spring feeding the lake. Both DGGE and molecu-
lar clone library results showed characteristic differences
between the bacterial communities inhabiting these sedi-
ment samples.
In the case of sample AS-1, dominance of Acidobac-
teria was revealed (Fig. 4). The most abundant ARDRA
group was closely related to Granulicella arctica (Män-
nistö et al. 2012). Occurrence of members of genus Gran-
ulicella is frequent in Polar Regions, several species were
Extremophiles (2016) 20:603–620 611

99 AS-2-10 (LN929580) / 7 clones

99 Uncultured bacterium clone A1 e2 (HQ317061) / Acid rock drainage (Antarctica)
AS-1-86 (LN929574) / 2 clones
43
100 Uncultured bacterium clone V6 44 (JF267704) / Coal mine heap, Sokolov (Czech Republic)

AS-13-30 (LN929664) / 2 clones

86 100 Uncultured bacterium clone 4.4.9 (EU528206) / Kentucky lake sediment (USA) Betaproteobacteria
100 AS-5-55 (LN929611) / 5 clones
98
Uncultured bacterium clone GA091 (EU636041) / Collins glacier (Antarctica)
86 AS-10-7 (LN929621) / 3 clones
100 Uncultured bacterium clone 1250 (FM209333) / Sand soil, Negev desert (Israel)

AS-5-45 (LN929607) / 1 clone

100 Uncultured bacterium clone reservoir-108 (JF697489) / Stream of Dianchi lake (China)
98 AS-13-13 (LN929656) / 2 clones
84
100
AS-13-75 (LN929685) / 1 clone
54 Uncultured bacterium clone Alchichica AL52 2 1B 85 (JN825498) / Alchichica alkaline lake (Mexico)
AS-10-71 (LN929643) / 4 clones
100 Uncultured bacterium clone SR-O-03-32 (AM905315) / High-Arctic intertidal beach sediment (Norway) Gammaproteobacteria
45

96
AS-10-84 (LN929648) / 1 clone
58 100 Uncultured bacterium clone KD2-14 (AY218573) / Penguin dropping sediment, Ardley Island (Antarctica)
AS-13-36 (LN929667) / 1 clone
100 Uncultured bacterium clone FFCH8865 (EU134765) / Soil of mixed grass prairie (USA)
100 AS-13-46 (LN929674) / 1 clone
AS-13-19 (LN929659) / 1 clone
16
94 Uncultured bacterium clone TX1A 146 (FJ152698) / Alkaline saline soil, former Lake Texcoco (Mexico) Alphaproteobacteria
76
AS-1-14 (LN929555) / 1 clone
100 Uncultured bacterium clone JFJ-ICE-Bact-04 (AJ867748) / Melted-ice water (Switzerland)
100 AS-2-5 (LN929578) / 10 clones
AS-2-3 (LN929577) / 15 clones Firmicutes
91 Uncultured bacterium clone Central-Bottom-cDNA clone89 (HE604029) / Acidic lignite mine lake (Germany)
100 AS-14-91 (LN929721) / 5 clones

17 100 Uncultured bacterium clone LEGE 07319 (HM217045) / Estuary (Portugal) Cyanobacteria
AS-14-6 (LN929694) / 1 clone
100 Uncultured bacterium clone SINH475 (HM128037) / Xiaochaidan lake (Tibet)
36 100 AS-13-63 (LN929678) / 1 clone Chloroflexi
Uncultured bacterium clone FCPT687 (EF515885) / Grassland soil (CA USA)
54 AS-1-74 (LN929570) / 3 clones
97 AS-1-44 (LN929562) / 6 clones
100 TM7
AS-1-63 (LN929567) / 1 clone
82
Uncultured bacterium clone C129 (AF507687) / pinyon-juniper forest soil, Arizona (USA)
100 AS-5-31 (LN929605) / 1 clone
99 Uncultured bacterium clone P3 (DQ351728) / Soil of LaGorce mountains (Antarctica) Bacteroidetes
AS-13-60 (LN929676) / 1 clone
100 Uncultured bacterium clone 20m-41 (GU061294) / Intertidal beach seawater, Yellow Sea (Korea)
100 AS-5-9 (LN929599) / 2 clones
Uncultured bacterium clone 5B 04E (JX098559) / 6000 meters elevation mineral soils (Atacama desert)
100
99 AS-5-61 (LN929613) / 2 clones
Uncultured bacterium clone AK4AB2 04A (GQ396928) / Recently deglaciated soil, Mendenhall glacier (USA) Gemmatimonadetes
74 99 AS-13-72 (LN929682) / 1 clone
100 Uncultured bacterium clone KC-4 (EU421850) / Soil under a glacier, Lahaul-spiti Valley (Indian Himalaya)

AS-13-5 (LN929655) / 1 clone

24
100 Uncultured bacterium clone BN23 (HQ190281) / Zhongynan oil field (China)
100 AS-1-80 (LN929573) / 2 clones
Uncultured bacterium clone P23 (DQ351736) / Soil of LaGorce mountains (Antarctica)
100 AS-2-12 (LN929582) / 17 clones
89 Actinobacteria
Uncultured bacterium clone OY07-C082 (AB552368) / Volcanic ash deposit (Japan)
99 AS-1-32 (LN929560) / 1 clone
100 Uncultured bacterium clone OY07-C186 (AB552456) / Volcanic ash deposit (Japan)
22 71 AS-5-92 (LN929618) / 2 clones
100 Uncultured bacterium clone A01 SB2A (FJ592652) / High-altitude warm fumarole soil, Socompa volcano (Andes) Verrucomicrobia
AS-13-73 (LN929683) / 4 clones
92 Uncultured bacterium clone Par-s-7 (EF632909) / High-altitude freshwater sediment (Chilean Altiplano)
99 AS-13-41 (LN929671) / 1 clone
84 Uncultured bacterium clone RB41(Z95722) / n.d.
AS-13-77 (LN929686) / 2 clones
100
51 99 Uncultured bacterium clone GTM2314fO9 (JN615809) / Microbial mat from lava cave (Portugal)

AS-10-44 (LN929634) / 2 clones

100 AS-5-50 ( LN929609) / 1 clone
61 Uncultured bacterium clone EME017 (EF127595) / Ice of Dry Valleys (Antarctica)
87 AS-2-56 (LN929590) / 1 clone Acidobacteria
72 100 AS-2-85 (LN929596) / 1 clone
AS-2-81 (LN929594) / 1 clone
65
89 Uncultured bacterium clone RT8-ant02-d03-W (JF807641) / Rio Tinto water (Spain)

AS-1-77 (LN929571) / 3 clones

100 90 Uncultured bacterium clone B146 (FJ466009) / Kilauea volcanic deposit (Hawaii)
AS-1-29 (LN929558) / 1 clone
95 Uncultured bacterium clone ERF-1A1 (DQ906050) / Rhizosphere on the bank of Rio Tinto (Spain)

0.02

AS-1: 6500 m (cold water)

AS-2: 6500 m (warm water)
AS-5: 5900 m (cold water)
AS-10: 5900 m (cold water)
AS-14: 4350 m (warm water)
AS-13: 3770 m (cold water)

13

612
described from tundra soils in Finland. Species G. arc-
tica was reported to be dominant in acidic soils, showing
growth at only 4 °C, and being tolerant to several freeze–
thaw cycles (Männistö et al. 2012). The second most
abundant major ARDRA group was related to environ-
mental clones from forest soils belonging to the candi-
date phylum TM7 (Dunbar et al. 2002). In AS-1 members
of Actinobacteria were only far relatives of three species,
Glaciihabitans tibetensis, isolated from glacial meltwater
affected sediment in Tibet (Li et al. 2014), the psychro-
philic (optimum at 1–15 °C) Alpinimonas psychrophila,
described from cryoconites of Rettenbach glacier, Aus-
tria (Schumann et al. 2012), and the halotolerant (<4 %
NaCl) Calidifontibacter indicus, described from a hot
spring in India (Ruckmani et al. 2011). Other molecular
clones from this library belonging to the phylum Actino-
bacteria showed the highest sequence similarity to uncul-
tured bacteria detected in soil samples from La Gorce
mountains, Antarctica (Aislabie et al. 2006) and from
volcanic sediments in Japan (Fujimura et al. 2012).
In sample AS-1, representatives of three classes of Pro-
teobacteria (Alpha-, Beta-, and Gammaproteobacteria)
were detected (Fig. 4). Members of Alphaproteobacte-
ria were related to molecular clones from mineral soil at
6000 m.a.s.l, in the Dry-Andes (Lynch et al. 2012). A
molecular clone was closely related to the mesophilic spe-
cies Rhodanobacter umsongensis (Kim et al. 2013) which
was also detected by DGGE analysis. Some molecular
clones of the sample AS-1 showed low similarity to molec-
ular clones from soil of Princess Elisabeth Station, Antarc-
tica (Peeters et al. 2011) and to species Mucilaginibacter
frigoritolerans a freeze-thaw cycle tolerating bacterium
(Männistö et al. 2010) within phylum Bacteroidetes.
In sample AS-2 more than 33 % of the clones showed
the highest similarity to environmental sequences belong-
ing to the phylum Firmicutes (Fig. 4). This phylum was
also present in the other warm sample AS-14. Phylum Act-
inobacteria was the second most abundant group. Molecu-
lar clones showed the highest similarity to environmental
sequences, e.g., from the formerly mentioned volcanic sed-
iment in Japan (Fujimura et al. 2012), the Mendenhall gla-
cier in the United States, and clones from the acidic river
Rio Tinto, Spain (García-Moyano et al. 2012). Molecular
clones of sample AS-2 belonging to Acidobacteria were
also similar to sequences from these acidic environments.
Representatives of the phylum Cyanobacteria were also
detected similar to the other warm-water sample AS-14.
13
Extremophiles (2016) 20:603–620
Comparison of the communities inhabiting the lake
sediments of Ojos del Salado and similar volcanic,
high‑altitude lake or permafrost sediments
Communities detected in the lake sediments of Ojos del
Salado consisted of sequences related mainly to psychro-
philic, acidophilic or halophilic microorganisms. The
composition of the bacterial communities was similar to
those reported from volcanic, high-altitude or permafrost
sediments (Hultman et al. 2015; Taş et al. 2014). In these
areas low organic material in soils, freeze-thaw cycles and
aridity are probably the main components of the environ-
mental stress which resulted in similar bacterial commu-
nity composition at phylum level. Proteobacteria, Act-
inobacteria and Bacteroidetes were the most frequently
detected phyla in almost all studied environments includ-
ing the samples from the area of Ojos del Salado, as well
(Table 3).
Regarding the number of phyla, each clone library from
the sediment samples of the lakes in the area of Ojos del
Salado contained members of at least 5 phyla (Table 3). It
is more than those found in similar high-altitude or perma-
frost-affected non-volcanic environments, for example in
the glacier melt water on Mount Everest (Liu et al. 2006),
permafrost soil of the Antarctic La Gorce mountain (Aisla-
bie et al. 2006) and Princess Elisabeth station, Antarctica
(Peeters et al. 2011).
In the present study, the most diverse bacterial com-
munities were detected in the samples from the lower
altitudes. Similarly diverse communities were only
reported from other active volcanic environments in the
Dry-Andes, e.g., Socompa volcano (Costello et al. 2009)
and Llullaillaco volcano (Lynch et al. 2012). Bacte-
rial community composition of samples AS-5 and AS-6
showed the highest similarity with those inhabiting
fumaroles and soils on Socompa volcano (Costello et al.
2009), while sample AS-2 was similar to a cold fumarole
on Socompa volcano (Costello et al. 2009) and a young
volcanic sediment from lower altitude (775 m) in Japan
(Fujimura et al. 2012). This suggests that in the case of
sample AS-2 acidification due to the volcanic activity
could be one of the main environmental stress factor,
while in case of the other samples (especially AS-1, AS-5
and AS-10) the low temperature owing to the presence of
permafrost and the freeze-thaw cycles could be the main
selective factors determining the bacterial community
compositions.
 Table 3  Comparison of bacterial phyla detected by clone libraries from the lake sediments of Ojos del Salado and other volcanic, high-altitude or permafrost environments
Origin of sample Sample type Elevation Aquificae Deinococcus- Chloroflexi Nitrospirae Chalditrix Cyanobac- Chlorobi Alphapro- Betaproteo- Gammapro-
(m.a.s.l.) Thermus teria teobacteria bacteria teobacteria

AS-1, Ojos del Sediment 6500 + + +

Salado, Dry
Andes (this
study)
AS-2, Ojos del Sediment 6500 + + +
Salado, Dry
Extremophiles (2016) 20:603–620

Andes (this
study)
AS-5, Ojos del Sediment 5900 +
Salado, Dry
Andes (this
study)
AS-10, Ojos del Sediment 5900 + + + + + +
Salado, Dry
Andes (this
study)
AS-14, Ojos del Sediment 4350 + + + + +
Salado, Dry
Andes (this
study)
AS-13, Ojos del Sediment 3770 + + + + +
Salado, Dry
Andes (this
study)
Simba crater Sediment 5870 + + +
lake, Andes
(Demergasso
et al. 2010)
Lake Namucuo, Sediment 4718 + + +
Tibetan plateau
(Xing et al.
2009)
Socompa vol- Cold fuma- 5824 + + + + +
cano, Andes role
(Costello et al.
2009)
Socompa vol- Warm fuma- 5824 + + +
cano, Andes role
(Costello et al.
2009)

13
613

614

Table 3  continued
Origin of sample Sample type Elevation Aquificae Deinococcus- Chloroflexi Nitrospirae Chalditrix Cyanobac- Chlorobi Alphapro- Betaproteo- Gammapro-

13
(m.a.s.l.) Thermus teria teobacteria bacteria teobacteria
Socompa vol- Sediment 5824 + + + +
cano, Andes
(Costello et al.
2009)
Llullaillaco Sediment 6330 + +
volcano, Andes
(Lynch et al.
2012)
La Gorce moun- Sediment 1800 + +
tain, Antarctica
(Aislabie et al.
2006)
Princess Elisa- Sediment nd
beth Station,
Antarctica
(Peeters et al.
2011)
Volcanic sedi- Sediment 775 + +
ment, Japan
(Fujimura et al.
2012)
Rongbuk glacier, Water 5140 + + +
morain lake,
Mount Everest
(Liu et al.
2009)
Glacier melt- Water 6350 +
water, Mount
Everest (Liu
et al. 2009)
Salar de Aguas Water 4200 + +
Calíentes,
Andes (Demer-
gasso et al.
2010)
Laguna Lejía, Water 4325 + + +
Andes (Demer-
gasso et al.
2010)
Laguna Vilama, Water 4650 + +
Andes (Farías
et al. 2009)
Extremophiles (2016) 20:603–620
 Table 3  continued
Origin of sample Deltapro- Epsilone Firmicutes Actinobac- Planctomy- Acidobac- Bacteroi- Verrucomi- Gemmati- WS5 TM7 OD1
teobacteria proteobac- teria cetes teria detes crobia monadetes
teria

AS-1, Ojos del + + + +

Salado, Dry
Andes (this
study)
AS-2, Ojos del
Extremophiles (2016) 20:603–620

+ + +
Salado, Dry
Andes (this
study)
AS-5, Ojos del + + + + + +
Salado, Dry
Andes (this
study)
AS-10, Ojos del + + + +
Salado, Dry
Andes (this
study)
AS-14, Ojos del + + + +
Salado, Dry
Andes (this
study)
AS-13, Ojos del + + + + + + +
Salado, Dry
Andes (this
study)
Simba crater + +
lake, Andes
(Demergasso
et al. 2010)
Lake Namucuo, + +
Tibetan plateau
(Xing et al.
2009)
Socompa vol- + + + + + +
cano, Andes
(Costello et al.
2009)
Socompa vol- + + + + + +
cano, Andes
(Costello et al.
2009)

13
615


Table 3  continued
616

Origin of sample Deltapro- Epsilone Firmicutes Actinobac- Planctomy- Acidobac- Bacteroi- Verrucomi- Gemmati- WS5 TM7 OD1
teobacteria proteobac- teria cetes teria detes crobia monadetes

13
teria
Socompa vol- + + + + + +
cano, Andes
(Costello et al.
2009)
Llullaillaco + + + + + + +
volcano, Andes
(Lynch et al.
2012)
La Gorce moun- + +
tain, Antarctica
(Aislabie et al.
2006)
Princess Elisa- + + +
beth Station,
Antarctica
(Peeters et al.
2011)
Volcanic + + +
sediment, Japan
(Fujimura et al.
2012)
Rongbuk glacier, + + +
morain lake,
Mount Ever-
est (Liu et al.
2009)
Glacier melt- +
water, Mount
Everest (Liu
et al. 2009)
Salar de Aguas +
Calíentes,
Andes (Demer-
gasso et al.
2010)
Laguna Lejía, + + + +
Andes (Demer-
gasso et al.
2010)
Extremophiles (2016) 20:603–620
Extremophiles (2016) 20:603–620 617

Conclusions

The results of this study demonstrated that sediments of

OD1

high-altitude lakes located in the Ojos del Salado volcano,

Dry-Andes maintain bacterial communities with decreasing
phylogenetic diversity along with the increasing altitude.
However, the relatively high difference of bacterial diver-
TM7

sity among the samples from the same altitude indicated

that both the extreme aridity and active volcanism influence
the bacterial community composition at the area of Ojos
del Salado. At phylum level, the phylogenetic diversity was
WS5

similar to other volcanic, high-altitude lake or permafrost

sediments from the world but at lower taxonomic levels,
monadetes

representatives of several unique and potentially novel taxa

Gemmati-

were revealed from these remote and unexplored habitats.

Verrucomi-

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