Microbial diversity and trophic components of two high altitude wetlands of the Chilean Altiplano

Abstract

This study examines the limnology and ecology of two high altitude wetlands, Lirima (19°51’24 S; 68°55’02 W; 4000 m asl) and Caya (20°37’21 S; 68°58’28 W; 3700 m asl), located in the Chilean Altiplano. Both wetlands are formed by the evaporitic remnant basins of paleolakes which occupied an extensive area of what today is known as the Altiplano. These systems have a negative hydrological balance, receiving their water from groundwater, snow melt and limited seasonal rains. An ongoing negative water balance and the sediment characteristics in the region have accelerated the salinization process in these systems, as shown by their present physicochemical characteristics. Nutrient values were typical of mesotrophic to eutrophic systems. The ionic content classifi es Lirima as a sodium sulfated wetland and Caya as a calcium chloride one. Conductivity values ranged between 778 μS/cm at Lirima to 2100 μS/cm at Caya, and were refl ected in the differences in biodiversity found in these systems. The Lirima wetland supports a population of the endemic fi sh Orestias aff. agassii found in several Evolutionary Significant Units (ESU) across the region. Microbial diversity in the water column was characterized by the presence of 5 bacterial phyla and related genera (e.g. Psychrobacter, Bacillus, Eryhtobacter, Halomonas). We present information on several key ecosystem components including macrophytes, plankton, benthos, fish and birds. This descriptive paper highlights the unusual limnological and biological characteristics of high altitude wetlands and highlights the importance of describing their biological communities across levels of organisation (e.g. microbial through to higher vertebrates) as well as their functional role, interactions and sensitivity to changes in water availability.

Full text

45
Microbial diversity and trophic components of two high altitude
wetlands of the Chilean Altiplano
Diversidad microbiana y componentes trófi cos de dos humedales de altura del altiplano
chileno
SERGIO SCOTT1, CRISTINA DORADOR2,3*, JUAN PABLO OYANEDEL1, IGNACIO TOBAR1, MARTHA HENGST2,3,
GIANNINA MAYA2, CHRIS HARROD4,5 & IRMA VILA1
1Departamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile. E-Mail:
limnolog@uchile.cl, Phone: +56 2 29787320. Fax: +56 2 22727363.
2Departamento de Biotecnología, Facultad de Ciencias del Mar y Recursos Biológicos y Laboratorio de Complejidad Microbiana
y Ecología Funcional, Instituto Antofagasta, Centro de Bioinnovación, Universidad de Antofagasta. *E-mail: cristina.dorador@
uantof.cl, Phone: +56 55 2657701. Fax: +56 55 2637804.
3Centro de Bioingeniería y Biotecnología, Universidad de Antofagasta, Angamos 601, Antofagasta, Chile.
4Instituto Alexander von Humboldt, Facultad de Ciencias del Mar y Recursos Biológicos, Universidad de Antofagasta, Angamos
601, Antofagasta, Chile.
5Queen Mary University of London, School of Biological and Chemical Sciences, Mile End Road, London, E1 4NS, UK.
ABSTRACT
This study examines the limnology and ecology of two high altitude wetlands, Lirima (19°51’24 S; 68°55’02 W; 4000 m
asl) and Caya (20°37’21 S; 68°58’28 W; 3700 m asl), located in the Chilean Altiplano. Both wetlands are formed by the
evaporitic remnant basins of paleolakes which occupied an extensive area of what today is known as the Altiplano. These
systems have a negative hydrological balance, receiving their water from groundwater, snow melt and limited seasonal
rains. An ongoing negative water balance and the sediment characteristics in the region have accelerated the salinization
process in these systems, as shown by their present physicochemical characteristics. Nutrient values were typical of
mesotrophic to eutrophic systems. The ionic content classifi es Lirima as a sodium sulfated wetland and Caya as a calcium
chloride one. Conductivity values ranged between 778 μS/cm at Lirima to 2100 μS/cm at Caya, and were refl ected in the
differences in biodiversity found in these systems. The Lirima wetland supports a population of the endemic fi sh Orestias
aff. agassii found in several Evolutionary Signifi cant Units (ESU) across the region. Microbial diversity in the water column
was characterized by the presence of 5 bacterial phyla and related genera (e.g. Psychrobacter, Bacillus, Eryhtobacter,
Halomonas). We present information on several key ecosystem components including macrophytes, plankton, benthos, fi sh
and birds. This descriptive paper highlights the unusual limnological and biological characteristics of high altitude wetlands
and highlights the importance of describing their biological communities across levels of organisation (e.g. microbial
through to higher vertebrates) as well as their functional role, interactions and sensitivity to changes in water availability.
KEYWORDS: Altiplano, biodiversity, endemism, evolutionary signifi cant units, salinity.
RESUMEN
Este estudio examina la limnología y ecología de dos lagos de altura, los humedales de Lirima (19°51’24 S; 68°55’02
W; 4000 m asl) y Caya (20°37’21 S; 68°58’28 W; 3700 m asl) que están ubicados en el Altiplano Chileno, representando
cuencas evaporíticas remanentes de antiguos paleolagos los cuales ocuparon una amplia zona en lo que hoy conocemos
como Altiplano. Estos sistemas tienen un balance hídrico negativo, recibiendo su agua desde fuentes freáticas, derretimiento
de nueves y escasas lluvias estacionales. Las características del suelo junto con la demanda de agua en la región han
acelerado el proceso desalinización en estos sistemas siendo refl ejado en las características químicas actuales. Los valores
de nutrientes fueron típicos de sistemas mesotrófi cos a eutrófi cos. El contenido iónico clasifi ca Lirima como un humedal
sodio sulfatado y el humedal de Caya como un humedal de cloruro de calcio. Los valores de conductividad fl uctuaron entre
778 μS/cm en Lirima a 2100 μS/cm en Caya, refl ejándose en las diferencias de biodiversidad encontrada en estos sistemas.
El humedal de Lirima conserva el pez endémico Orestias que representa Unidades Evolutivas Signifi cativas (ESU) en
la región. La diversidad microbiana en muestras de agua estuvo caracterizada por la presencia de 5 fi lo bacterianos y
géneros relacionados (e.g. Psychrobacter, Bacillus, Eryhtobacter, Halomonas). Los componentes trófi cos estudiados
Gayana 79(1): 45-56, 2015. ISSN 0717-652X

Gayana 79(1), 2015
46
INTRODUCTION
The Chilean Altiplano, with an average altitude of over
3000 meters above sea level, includes a large and varied
number of freshwater ecosystems within its boundaries
from its northern (18°S, Lake Chungará) to southern limits
(27°S, Lake Negro Francisco). To date, 50 different closed
hydrographic basins have been identifi ed, which together
present signifi cant variation in physicochemical (e.g.
ionic concentrations) and geomorphologic characteristics
(Risacher et al. 2003a, 2003b; Moreno et al. 2009). These
systems are internationally important but are largely
undescribed, limiting our understanding of them and the
ability of managers and scientists to protect them and the
goods and services they provide from human impacts.
The evaporitic basins of the Chilean Altiplano originate from
paleolakes that occupied the extensive area of what today
is known as the Altiplano and the Atacama Desert (Chong
1988; Marquet . 1998). Today, these systems have a negative
hydrological balance, with water loss by evaporation
outweighing hydrological inputs via groundwater, snow
melt and limited seasonal rains (Aceituno 1997; Salazar
1997; Keller & Soto 1998).
Studies of the taxa inhabiting aquatic ecosystems in the
Altiplano have raised important questions with regard to
their evolutionary history, biogeographic relationships and
evolutionary adaptations to the signifi cant abiotic stresses
that vary at daily, seasonal and interannual scales (e.g.
temperature, UV radiation, water availability). In shallow
lentic systems in the Altiplano, known locally as bofedales,
this has resulted in the repeated presence of genetically
distinct and locally adapted Evolutionary Signifi cant Units
(ESU) in different taxa, for instance the fi sh Orestias aff.
agassii Valenciennes (Vila et al. 2010).
Water availability is key to the function and maintenance
of life in these high altitude freshwater ecosystems. This
extends from the varied primary producer communities
that include contributions from fringing macrophytes,
microalgae and microbial mats (Mitsh & Gosselink 2000;
Dorador et al. 2008; Riveros et al. 2012) that are often
exposed to desiccation, through to the higher taxa that
consume and transfer energy and nutrients as part of the
foodwebs such as fi sh (Vila et al. 2013).
A series of recent studies have highlighted that high altitude
wetlands have unusual microbial communities (Bacteria
and Archaea) that include the presence of previously
undescribed lineages of Bacteria (Dorador et al. 2013).
These taxa participate actively in different biogeochemical
cycles (Dorador et al. 2008a) and exhibit marked temporal
and spatial variation according to gradients of salinity,
temperature and solar radiation (Dorador et al. 2008b;
2010). To date, there has been little consideration of the
trophic role of microbial taxa in these ecosystems.
Across the Altiplano, the majority of aquatic ecosystems
remain undescribed, limiting our ability to understand
their ecological importance and sensitivity to human
impacts. Here, we provide a fi rst description of microbial
diversity and key trophic components of two undescribed
and contrasting wetlands (Lirima and Caya), located at
high altitude in the Tarapaca Region of northern Chile.
These ecosystems, like other wetlands located across the
Chilean Altiplano, are subject to elevated risk due to high
and increasing water demand and ongoing climate change
(Valdés-Pineda et al. 2014): this work therefore contributes
to the basic knowledge of threatened aquatic environments
in northern Chile.
MATERIALS AND METHODS
DESCRIPTION OF STUDY SITES
We examined two high-altitude wetlands: Lirima (19°51’24
S; 68°55’02 W; 4000 m asl) and Caya (20°37’21 S; 68°58’28
W; 3700 m). Both systems are shallow remnants of earlier
deeper lakes from the Tarapacá Ravine area. The Tarapacá
Ravine rises in the Andean highlands where annual inputs
of water are not greater than 150 mm/year (Aceituno 1997,
Risacher et al. 2003). The climatic regime of the Tarapacá
Ravine is pluvial, with higher freshets during summer due to
summer rains in the Altiplano. The Tarapacá Ravine system
receives waters from the Coscaya River and then forms the
Caya wetland. Further downstream, the system drains the
Lirima plains where it forms the Lirima wetland. This river
incluyeron macrófi tas, plancton, bentos, peces, anfi bios y aves. Este artículo descriptivo destaca las inusuales características
limnológicas y biológicas de los humedales de altura poniendo atención a la importancia de describir comunidades en
distintos niveles de organización biológica (desde tapetes microbianos hasta vertebrados superiores), pero también sus
funciones, interacciones y sensibilidad a cambios en la disponibilidad de agua.
PALABRAS CLAVE: Altiplano, biodiversidad, endemismo, unidades evolutivas signifi cativas, salinidad.

47
Limnology of Lirima and Caya wetlands: SERGIO SCOTT ET AL.
presents higher freshets during winter and summer due to
winter and summer rains (June–July and January-March)
(MOP-DGA, CADE-IDEPE 2004) (Figs. 1 and 2).
Aquatic ecosystems in the region are commonly used as
sources for water supply for industrial processes such as
mining and agriculture. The Caya wetland currently has 11
applications to extract water for the mining industry and 3
existing abstraction points which are likely affecting both
water levels of the wetland and its function. In contrast,
Lirima wetland has no existing applications for water
extraction request to date (Morales, 2011).
In order to characterise the different systems, we used a
Geographical Information System (GIS) approach using a
number of different GIS (ENVI 4.4, IDRISI ANDES 15.01,
ARC GIS 9.3 & ARC View 3.3). Satellite images were
obtained from the Earth Science Data Interface (ESDI) of
the Global Land Cover Facility, University of Maryland,
USA, with two views from 1999 and 2003 from the Landsat
7 satellite, sensor ETM+ with 30 meter spatial resolution.
During November 2009 we made a series of measurements
of physicochemical and biological characteristics in the
Caya and Lirima wetlands. Chemical and biological
variables where obtained from replicated water samples.
Due to the diffi culties involved in accessing wetlands in
this region, and the stresses involved with reduced water
availability, these observations were made during a period
with minimum water fl ow (Maximum water depth in Caya
= 0.3 m; Lirima = 0.8 m) with the aim of characterizing the
system under extreme conditions, allowing the identifi cation
of the key abiotic factors that drive ecosystem function
under such conditions.
PHYSICAL AND CHEMICAL VARIABLES
Water temperature was measured (±0.1°C) with digital
Hanna thermometer; conductivity and total dissolved solids
(TDS) were measured with a VWR portable conductivity
meter, and pH with a portable WTW pH meter (± 0.1 pH
unit). Salinity (NaCl) was measured (± 0.1) with a portable
VWR salinometer. Dissolved oxygen concentrations
were estimated using the Winkler method (APHA, 2001),
and percentage saturation was estimated using values
of dissolved oxygen, altitude and temperature. Total
phosphorous (TP) and total nitrogen (TN) were measured
according to Mühlhauser et al. (1986). Sulfates (SO4)
were measured with the turbidity method of Gölterman
et al. (1978). Bicarbonates (HCO3), carbonates (CO3) and
the cations Na, K, Ca and Mg were measured by atomic
absorption (APHA 2005).
BIOLOGICAL VARIABLES
Bacterial abundance was estimated from water samples
using DAPI staining (Porter & Feig, 1980). Samples
were fi xed in the fi eld with 2% formaldehyde, fi ltered in
polycarbonate fi lters with 0.22 μm pore size and posteriorly
analyzed by epifl uorescence microscopy (Olympus BX51).
Culturable bacterial diversity was determined using mineral
media using acetate and DMSO as energy sources (Atlas
1995). The samples used to determine the diversity of
culturable bacteria were obtained from water samples where
fi eld temperatures fl uctuated between 21-25 ºC collected
from the Lirima wetland. Isolates were identifi ed through
their 16S rRNA sequences, which were obtained by PCR
and direct sequencing (Dorador et al. 2009). Their closest
relatives were determined by BLAST search (http://www.
ncbi.nlm.nih.gov/blast) and the classifi er tool in RDP II
(http://www.cme.msu.edu/rdp). Chlorophyll a concentration
was measured according to Montecino & Cabrera (1982).
Phytoplankton samples were obtained with a net 40 μm
size and fi xed with Lugol. In the laboratory, phytoplankton
were identifi ed and counted following the Utermöhl method
using an inverted microscope (Olympus CK2) according to
Villafaña & Reid (1995), in 10 ml and 25 ml sedimentation
cells during 24 hrs and 48 hrs respectively. Rivera et al.
(1982), Parra & Bicudo (1995) and Diaz & Maidana (2005)
were used for taxonomic identifi cation of phytoplankton to
the genus level. Macrophytes were collected and identifi ed
according to Ramírez and San Martín (2006), with Point
Quadrat methodology, with a sampling each two meters at
ten points using an inter-point distance of 10 cm.
FIGURE 1. Map of the Altiplanic region, northern Chile. Study sites.
FIGURA 1. Mapa de la Región Altiplánica, norte de Chile. Sitios de
Estudio.

Gayana 79(1), 2015
48
FIGURE 2. Classifi cation of land cover in Lirima and Caya based on images of Landsat ETM+, from 1999.
FIGURA 2. Clasifi cación de la cobertura de suelo en Lirima y Caya basados en imágenes Landsat ETM+, de 1999.
Zooplankton samples were obtained by fi ltering 10 L of
water of the water column with a zooplankton net 120 μm
size and fi xed with 10% buffered formaldehyde. In the
laboratory, they were counted in a Bogorov cell (Wetzel &
Likens 1991). For taxonomic identifi cation the taxonomic
keys of Araya & Zuñiga (1985) were used. Benthic fauna
samples were obtained with a Surber sampler and identifi ed
according to Fernández & Domínguez (2001) and Figueroa
et al. (2003). Fish and amphibians were captured with
electrofi shing equipment and identifi ed in the laboratory
according to Vila et al. (2006). The avian communities using
the wetlands were assessed over one hour in each system at
midday, counted and identifi ed in the fi eld.
RESULTS
Landcover in the catchment of the study wetlands is
heterogeneous and characterized by seven distinct cover
types corresponding to: i) Xerophytic grassland, ii) Bofedal
(low vegetation forming “cushions”) iii) Stream (superfi cial
river bed), iv) Bush (50 to 150 cm shrub formation), v) Naked
soil, vi) Anthropological relicts and vii) Hydrothermal
waters, (Ahumada & Faundez, 2009) (Figs. 1 and 2).
TABLE 1 details the physicochemical characteristics recorded
from the Lirima and Caya wetlands. Water temperature was
within the typical range of Altiplano systems, with high
values during the day due to the diurnal heating of the shallow
high Andean systems (>20° C). Conductivity analysis
showed values of 778 μS/cm at Lirima, characteristic of
freshwater systems (Wetzel 2001). Conductivity was higher
in the Caya wetland, reaching 2100 μS/cm. Values of pH
were basic, fl uctuating between 8.3 and 8.7 due to the salt
concentration of these systems.
Oxygen values were high for high altitude systems,
especially in Caya, and varied between 10 and 14 mg/L at
Lirima and Caya respectively. In the Lirima wetland, the
oxygen saturation percentage reached 100% and at Caya it
increased to 130%. TP values were especially high in Lirima
(169.9 μg/ L) which corresponds to high productivity or
eutrophic systems. Caya presented lower values of TP
(67.8 μg/L-1), in the range of mesotrophic systems (Ryding

49
Limnology of Lirima and Caya wetlands: SERGIO SCOTT ET AL.
TABLE 1. Physical and chemical variables of the Caya and Lirima
systems (average of replicated samples).
TABLA 1. Variables físicas y químicas de los sistemas de Caya y
Lirima (promedio de réplicas de muestras).
SITE CAYA LIRIMA
Depth (m) l l
Total nitrogen (μg/L) 2421.6 1631.9
Total Phosphorus (μg/L) 67.8 169.9
Cl- (mg/L) 450 67
HCO3
-
(mg/L) 79.3 26.5
CO3
-2
(mg/L) 0.0 15.0
Na+ (mg/L) 188.3 91.1
K+ (mg/L) 18.6 16.3
Ca+2 (mg/L) 261.1 60.9
Mg+2 (mg/L) 40.1 9.2
Chl a (mg/L) 5.4 2.9
Dissolved oxygen (mg/L) 12.5 8.9
SO4
-2 (mg/L) 172.1 82.5
Temperature (°C) 24.1 22.1
Conductivity (μS/cm) 2100 778
pH 8.3 8.7
& Rast 1992). Values of TN were relatively high and
correspond to systems from meso- to eutrophic in Lirima
and Caya, reaching 1631.6 and 2421.6 μg/L, respectively
(Ryding & Rast 1992; Wetzel 2001). Chloride values
were high in Caya (450 mg/L) and lower in Lirima with
67 mg/L. Sulfate values were 82.5 mg/L and 172 mg/L at
Lirima and Caya, respectively. Bicarbonate values were
low and fl uctuated between 26.5 mg/L and 79.3 mg/L for
Lirima and Caya, respectively. CO3
-2 values were 15 mg/L
at Lirima and under the detection limit at Caya. According
to the cation concentrations, Lirima represents a system
with predominance of sodium (Na<Ca<K<Mg). Caya,
by contrast, has dominance of calcium (Ca<Na<Mg<K).
Figure 3 summarizes the anionic and cationic values for
both systems in a Maucha diagram (Wetzel 2001), which
highlights the proportionality of ion concentrations in both
wetlands.
Bacteria present in Lirima water samples were analyzed by
DAPI, estimating an average concentration of 3×104 cell/
ml. Microbial cells were mainly grouped in a fi lamentous
matrix. In the hydrothermal waters of Lirima wetland, the
presence of highly pigmented microbial mats was common.
In total we analyzed 30 16S rRNA gene sequences from
61 bacterial isolates. The isolates were members of
Gammaproteobacteria (43%), Alphaproteobacteria (13%),
Firmicutes (37%), Actinobacteria (3.3%) and Bacteroidetes
(3.3%). Most of the isolates exhibited high sequence
identity (>98%) with the following genera: Psychrobacter,
Halomonas, Aeromonas, Pseudomonas, Paracoccus,
Brevundimonas, Rhizobium, Exiguobacterium, Bacillus,
Planococcus, Dietzia and Chryseobacterium (Fig. 4). Most
of these taxa use different organic carbon sources and also
exhibit adaptations to thrive under extreme environmental
conditions such as low temperature (Psychrobacter,
Chryseobacterium), high salt concentration (Halomonas)
and desiccation (Bacillus) (Table 2).
Chlorophyll a values were in direct accordance with the
amount of dissolved nutrients in these wetlands, especially
TN in Caya. These values fl uctuated between 5.4 μg/L
in Lirima and 2.9 μg/L in Caya, which corresponds to
mesotrophic systems (Wetzel 2001). As in the majority of
the Altiplano wetlands, microalgae were mostly represented
by yellow silica algae or diatoms (Diaz & Maidana 2005).
At Caya there was a higher abundance of cyanobacteria
in correspondence with higher values of total nitrogen.
Their abundance reached a total of 135.6 and 108.9
units/L in Caya and Lirima, respectively (Table 3). The
macrophyte communities at both locations were limited:
at Lirima macrophytes were dominated by “water pine“,
Myriophyllum sp., followed by Lilaeopsis sp.; at Caya
only Lilaeopsis, Patosia and Ranunculus were recorded
(Table 3). Zooplankton abundance was low and dominated
by Cladocera in both wetlands. Ostracoda followed in
abundance, (Table 4). Benthic fauna in both wetlands were
mostly composed of the crustacean genus Hyalella, which is
generally very abundant in shallow waters associated with
macrophytes (Table 5).
The bird community of the two systems included 15
species, including the Andean seagull Larus serranus
(Tschudi) classifi ed as a rare species, and the vulnerable
species Andean goose Chloephaga melanoptera (Eyton),
(Table 6). Fish were only recorded Lirima where karachi
Orestias aff agassii Valenciennes were present; this genus
is endemic to the Altiplano area and all of its species are
classifi ed as endangered, due to their low population
number and restricted distribution according to the current
Environmental Ministry species classifi cation rules (http://
www.mma.gob.cl/clasifi cacionespecies/) (Table 6).

Gayana 79(1), 2015
50
TABLE 2. Phylogenetic affi liation of bacterial isolates obtained from water samples of Lirima wetland based on 16S rRNA gene sequence
identity with GenBank and Ribosomal Database Project (RDP).
TABLA 2. Afi liación fi logenética de aislados bacterianos obtenidos desde muestras de agua del humedal de Lirima basados en la identidad
de secuencia del gen ribosomal ARN 16S con las bases de datos GenBank y Ribosomal Database Project (RDP).
Bacterial
strain
First hit in BlastN (accesion number) (% coverage/ % sequence
identity)
Phylogenetic affi liation (RDP) (Phyla/
Subphyla, Genus)
9-11 Psychrobacter sp. BSw20963B (GU166134) (99/96) Gammaproteobacteria, Psychrobacter
L8-11 Psychrobacter marincola strain J82 (JX976309) (100/100) Gammaproteobacteria, Psychrobacter
L52-32 Halomonas sp. SB135_6 (EU308331) (100/99) Gammaproteobacteria, Halomonas
L5-11 Aeromonas hydrophila pc104A (CP007576) (95/99) Gammaproteobacteria, Aeromonas
L33-41 Clone 661206 (DQ404928) (100/99) Gammaproteobacteria, Pseudomonas
L66-21 Erythrobacter sp. JL660 (EF512713) (100/99) Alphaproteobacteria, Erythrobacter
L37-41 Paracoccus marcusii strain zzx35 (KJ009431) (100/99) Alphaproteobacteria, Paracoccus
L51-21 Clone CT1C2BB10 (JQ427853) (100/99) Alphaproteobacteria, Brevundimonas
L48-41 Rhizobium sp. AB3 (KC870058) (100/99) Alphaproteobacteria, Rhizobium
L11-11 Exiguobacterium sp. Pb-WC11087 (JX913841) (100/99) Firmicutes, Exiguobacterium
L2-11 Bacillus sp. 58B112Y11 (KC815825) (100/99) Firmicutes, Bacillus
L47-MH Clone QNSW58 (FJ384526) (100/99) Firmicutes, Bacillus
L39-42 Clone ncd2231c10c1 (JF193171) (100/100) Firmicutes, Sinobaca
L46-12 Clone QNSW58 (FJ384526) (100/99) Firmicutes, Bacillus
L6-11 Clone QNSW58 (FJ384526) (100/99) Firmicutes, Bacillus
L21-22 Dietzia psychralcaliphila strain DSM 44820 (FJ468331) (99/99) Actinobacteria, Dietzia
L62-21 Chryseobacterium sp. L2 (KF358274) (100/99) Bacteroidetes, Chryseobacterium
TABLA 3. Abundancia de microalgas en Lirima y Caya (unidades por litro) (promedio de muestras replicadas). Número de individuo/Litro
en los sistemas de Lirima y Caya (*ausente) (promedio de muestras replicadas).
MICROALGAE LIRIMA
units/L
CAYA
units/L ZOOPLANKTON LIRIMA
ind/L
CAYA
ind/L
Diatoms Copepoda
Cocconeis 14.7 25.6 Ciclopoidea
Navicula 65.4 71.6 Eucyclops serrulatus 0.1 *
Synedra 18.2 24.0 Harpacticoidea * 0.4
Cimatopleura 2.1 1.6 Cladocera
Nitzchia 2.0 2.0 Alonella excisa * 0.3
Fragilaria 2.4 3.6 Chydorus sphaericus 0.4 *
Surirella 1.2 1.6 Rotifera
Gomphonema 2.0 2.0 Keratella cochlearis 0.1 *
Cyanobacteria Rotifer spp 1 0.1 *
Anabaena 0.9 2.8 Ostracoda 0.2 0.8
Desmids * 0.8
Total 108.9 135.6
TABLE 3. Abundance of microalgae in Lirima and Caya (Units per L) (average of replicated samples). Number of individuals/L of
zooplankton in the Lirima and Caya systems (* absent) (average of replicated samples).

51
Limnology of Lirima and Caya wetlands: SERGIO SCOTT ET AL.
FIGURE 3. B: Maucha diagrams characterising the anionic and cationic content of the wetlands of Caya and Lirima.
FIGURA 3. B: Diagramas de Maucha con el contenido anionico y catiónico de los sistemas de Caya y Lirima.
FIGURE 4. Phylogenetic tree inferred from partial 16S rRNA gene sequences (~900 bp) from isolates bacterial phylotypes of Lirima wetland.
The tree was constructed using maximum likelihood. Phylotypes are highlighted in bold and the number of isolates per phylotypes are
shown in brackets. The evolutionary distance was inferred using general time reversible method. Bootstrap values over 50% are shown.
Scale bar indicate 0.5 site substitutions. Halorubrum vacuolatum was used as outgroup.
FIGURA 4. Árbol fi logenético inferido de secuencias parciales del gen 16S del rRNA (~900 pares de bases) de fi lotipos bacterianos aislados
del humedal de Lirima. El árbol fue construido usando Máxima Verosimilitud. Filotipos son resaltados en negrita y el número de aislados
por fi lotipo son mostrados en paréntesis. La distacia evolutiva fue inferida usando el método general de tiempo reversible. Los valores de
Bootstraps mostrados son sobre el 50%. La barra de escala indica 0.5 sustituciones por sitio. Halorubrum vacuolatum fue utilizado como
grupo externo.

Gayana 79(1), 2015
52
TABLA 6. Presencia de aves asociadas a los sistemas de Lirima y Caya (×: presencia)
COMMON NAME SCIENTIFIC NAME LIRIMA CAYA CONSERVATION STATE
Red-backed sierra fi nch Phrygilus dorsalis (Cabanis) ××
Blue-and-white swallow Pygochelidon cyanoleuca (Lafresnaye y
D’Orbigny) ××
Andean gull Larus serranus (Tschudi) ×R
Flamingo Not identifi ed ×
Chilean teal Anas fl avirostris oxyptera (Meyen) ×
Lesser rhea Pterocnemia pennata tarapacensis Chubb ××
Mountain caracara Phalcoboenus megalopterus (Meyen) ××
Andean Condor Vultur gryphus (Linnaeus) ×V
Puna tinamou Tinamotis pentlandii (Vigors) ×V
Andean goose
Chloephaga melanoptera (Eyton)
×
×
V
Puna ibis Plegadis ridgwayi (Allen) ×V
Crested duck Lophonetta specularioides alticola
(Ménégaux) ××
Greenish Bellow-fi nch Sicalis olivascens (Tschudi) ×
Rufous-naped ground-tyrant Muscisaxicola rufi vertex pallidiceps
(Hellmayr) ×
White-winged cinclodes Cinclodes atacamensis (Philippi) ×
Black siskin Carduelis atrata (Lafresnaye y D’Orbigny) ×
V= Vulnerable; R= Rare. V= Vulnerable; R= Rara.
TABLE 6. Presence of aquatic birds and others associated with the Lirima and Caya systems. (×: presence).
TABLE 5. Benthic macroinvertebrates recorded from the Lirima and Caya systems (individuals/m2).
TABLA 5. Fauna bentónica de los sistemas de Lirima y Caya (Individuos/m2).
CLASS ORDER FAMILY GENUS LIRIMA CAYA
Arachnoidea Acari Hydrozetidae Hydrozetes -391
Hydrobatidae Atractidella -19
- Undetermined species -2
Crustacea Amphipoda Hyalellidae Hyalella 83 158
Hirudinea -- --11
Insecta Coleoptera Elmidae Austrelmis -7
Diptera Chironomidae Chironomus -52
Hemiptera Corixidae --1
Odonata Coenagrionidae --4
Total individuals 83 645
TABLE 4. Percentage coverage of aquatic macrophytes.
TABLA 4. Vegetación de macrófi tas acuáticas.
LIRIMA CAYA
Plant genera Plant cover (%) Plant cover (%)
Myriophyllum 80
Lilaeopsis 30.7 46.0
Eleocharis 6.7 -
Oxychloe 5.3 -
Patosia - 49.3
Ranunculus - 39.3

53
Limnology of Lirima and Caya wetlands: SERGIO SCOTT ET AL.
DISCUSSION
The Lirima and Caya wetlands showed characteristic
features of shallow systems known in the region as
bofedales (Ramirez et al. 2002; Ahumada & Faundez
2009). Water levels were low in both systems refl ecting the
climatic characteristics of the region where rain is extremely
limited (mean = 54.4 mm/year), and is exacerbated by high
evaporation rates (Salazar 1997), which drive the chemical
and physical characteristics of these shallow water systems.
The high ionic concentrations recorded from the wetlands
likely refl ect the combined effects of climatic factors with
high availability of ions form the underlying sediments.
Sulfate values are relatively high for both systems, a
common feature in Altiplano environments (Risacher et al.
2003b; Marquez et al. 2009). These systems are particularly
sensitive to hydrological changes, which may affect nutrient
availability and diminish dissolved oxygen content due to
warming during the day, and increased salinity in both
water and sediments. Reductions in water volume following
extraction and evaporation results in increases in salinity
and nutrient concentrations, due to the concentration of
ions. Thus, in contrast to the wetlands typical of temperate
regions, these systems do not make a stepwise shift from
a clear water state to a turbid one (Scheffer & van Nes
2007); here, relatively minor hydrological changes may
lead to a shift in ecosystem state, leading to a loss of
species richness e.g. in higher taxa, refl ecting the limited
tolerance to salinity shown by the majority of freshwater
invertebrates and vertebrates. It has been reported that
spatial and temporal changes in the Huasco salar, located
40 km from these study sites, have resulted in a distinct
microbial community (Dorador et al. 2008b; 2010) with
likely consequences for higher trophic levels. Here, the
culturable bacterial diversity obtained was restricted to the
culture media used, nevertheless it was possible to describe
at least fi ve different bacterial phyla and 13 genera. The
isolates obtained are functionally important, due to their
role in recycling of organic matter using different sources
of energy (e.g. Psychrobacter, Pseudomonas), anoxygenic
photosynthesis (e.g. Brevundimonas) and nitrogen fi xation
(e.g. Rhizobium). Also the closest culturable relative to
the these taxa were previously isolated from different
extreme environments exhibiting adaptations to a range of
environmental stressors, as high salinity, cold temperatures
and high solar radiation (Table 2). The diverse phylum found
are in agreement with the main microbial groups found in
other inland environments (e.g Newton et al. 2011), but
with the clear infl uence of high salt concentrations as the
main factor modulating the aquatic culturable bacteria
from the two wetands. The prevalence of these taxa and
their metabolic role could have important consequences for
higher trophic levels, e.g. through provision of energy and
the maintenance of suitable abiotic conditions.
Sodium chloride concentrations in the Caya wetland appears
to be such to limit the presence of fi sh, as has been reported
by Keller & Soto (1998) for the Ascotán salar. The presence
and maintenance of suitable volumes of water as well as
fringing vegetation is of primary importance in Atiplanic
wetlands. Macrophytes are important due to their ability
to retain nutrients and sediments, especially during the
Altiplano rains, as well as the provision of substrate support
and habitat for macroinvertebrates and fi shes (Mitsch &
Gosselink 2000; Gulati et al. 2005). The formation of the
large salt pans in the Altiplano region was principally due to
the climatic changes that have occurred in the region over
geological time scales (Chong 1988). Nevertheless, during
the last century, the salinization process has accelerated,
largely due to increased water extraction. Here, water
demand typically from agriculture and mining activities
greatly outstrips water availability, resulting in decreased
levels in many aquifers (Valdés-Pineda et al. 2014). The
Altiplano is extremely sensitive to changes in the effective
humidity (rains minus evaporation). Also, small changes
in water reservoirs could produce important and amplifi ed
responses in the majority of the salt pans, by modifying
morphological variables, geomorphologic processes,
vegetation changes and other biogeochemical variations
(Grosjean & Veit 2005). Solar radiation in the study area
can be up to 20% greater than that recorded at sea level at
the same latitude (Aceituno 1997) reaching values up to
1500 W/m2 in the Altiplano (Piacentini et al. 2003). Solar
radiation varies both annually and across daily cycles,
producing abrupt changes in photosynthetic parameters
and community structure (Hernandez et al. 2012).
Phytoplanktonic photosynthesis inhibition has been studied
in Lake Titicaca, showing a high degree of adaptation of the
phytoplankton species to changes of the levels of radiation
PAR, UVA and UVB (Villafaña et al. 1999) and likely
operates in the wetlands studied here.
The two wetlands described here showed different
physicochemical characteristics: Lirima can be characterized
as a sodium sulfate wetland, and Caya as a calcium chloride
wetland. Although both systems have relatively elevated
nutrient concentrations, it appears that the difference in
salinities between the two is associated with a reduction
in biodiversity. This supports calls for conservation of
minimum water levels in such sensitive ecosystems to
maintain chemical quality (Risacher et al 2003b) suitable
for communities of microbes, plants and other taxa.
Cartography shows that Caya is a shallow stream and Lirima
is rather a pond with hydrothermal affl uent; both have
interesting relicts of former indigenous cultures (Villagrán
& Castro 2003). Lirima sustains a low-density population of
Orestias that has elevated biogeographical and evolutionary
importance, since these fi sh are the only remnants of
Altiplanic fi sh in this zone (Vila et al. 2010). The Lirima

Gayana 79(1), 2015
54
population is an Evolutionary Signifi cant Unit (ESU), due
to its reproductive isolation and putative adaptation to
local conditions and has conservational importance at both
international and national scales (Waples 1991; Morales et
al. 2011). The species of the genus Orestias are presently in
danger of extinction across the entire Altiplano region (Vila
et al. 2010). In this system we also observed the presence
of the amphibian Telmatobius, also an important Altiplano
genus (Veloso 2006) (Data not shown). Twelve species of
birds were observed at Lirima and ten in Caya, including the
lesser rhea (Pterocnemia pennata tarapacensis Chubb) and
fl amingos, as an example of the importance of these wetlands
as corridors for conservation of bird life in endangered and
vulnerable conservation states (Salaberry & Tabilo 1990).
This descriptive paper highlights the unusual limnological
and biological characteristics of high altitude wetlands in
the Chilean Altiplano. Due to their international and national
importance, and the ongoing and increasing demands for
water in the region, there is a strong and pressing need
for studies describing the physicochemical and biological
characteristics of high altitude aquatic ecosystems across
the Altiplano region. Such studies should look to not
only describe communities across levels of biological
organisation (e.g. microbial through to higher vertebrates)
but also their function, interactions and sensitivity to
changes in water availability.
ACKNOWLEDGEMENTS
CONAMA (Comisión Nacional de Medio Ambiente),
Chile. Fondecyt Regular Projects N°1080390, 11080228,
1110953 and 1140543, CONICYT (National Commission
for Scientifi c and Technological Research), Chile.
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