ARTÍCULOS CIENTÍFICOS - TÉCNICOS Rev. Bol. Ecol. y Cons. Amb.

25: 51-67, 2009

Banco de huevos de resistencia revela una alta riqueza

específica de cladóceros en charcos temporales altoandinos

Resting egg bank reveals high cladoceran species richness

in high-altitude temporary peat land pools

Jorge S Coronel1, Ximena Aguilera1, Steven Decleck2 & Luc Brendonck2

RESUMEN

El análisis del banco de efipias (huevos de dormancia) presente en el sedimento de los cuerpos de agua ha resultado ser una
herramienta útil para el estudio de la diversidad de cladóceros en sistemas permanentes de agua. Este método ha sido
inexplorado en sistemas temporales que experimentan un periodo seco durante una parte del año. En este estudio se evaluó
la riqueza de especies del ensamblaje de cladóceros (Branchiopoda, Crustacea) obtenido mediante la eclosión de efipias
provenientes del sedimento de 61 charcos temporales de la cordillera del Tunari en Bolivia. La eclosión de efipias reveló más
especies de cladóceros (total= 24; promedio= 6.7) que el número obtenido de los muestreos del ensamblaje activo (total= 21;
promedio= 4.6). En promedio, el análisis de las efipias contribuyó con 2.1 especies de cladóceros por charco. El número de
cladóceros (ensamblaje activo + análisis de efipia) alcanzó un total de 28 especies. Análisis de redundancia indicaron diferencias
significativas entre la composición del ensamblaje activo y el ensamblaje obtenido por eclosión de las efipias. La cobertura
vegetal fue la principal variable que explicó una variación en la estructura del ensamblaje activo de cladóceros, mientras que
para el ensamblaje obtenido de la eclosión de efipias estuvieron la clorofila-a, el grosor de la capa de sedimento, y la conectividad
entre charcos.

Palabras claves: Bolivia, Charcos temporales andinos, huevos de dormancia, zooplancton de los Andes.

ABSTRACT

The use of egg bank analysis proved a valuable approach to uncover hidden diversity in permanent lakes. The efficiency of
the method remains largely unexplored in temporary aquatic systems that remain dry for a variable part of the year. We assessed
species richness of the cladoceran assemblage (Branchiopoda, Crustacea) from the dormant egg bank of 61 temporary peat
land pools in the high Andes of Bolivia. The analysis of the dormant egg bank yielded more species (total: 24; mean per peat
land pool: 6.7) than snapshot samples from active communities taken previously (total: 21; mean per peat land pool: 4.6). On
average, the dormant egg bank resulted in the detection of 2.1 (45%) more cladoceran species per peat land pool than on the
basis of active cladoceran assemblages. The accumulated (active plus dormant) cladoceran species richness of the study peat
land pools mounted up to 28 species. RDA analyses indicated a significant difference in assemblage composition between
dormant and active samples. Different environmental variables explained variation in the structure of the dormant and active
cladoceran assemblages. Water plant cover significantly explained variation in active assemblages, while a model constructed
by chlorophyll-a, sediment thickness, and connectivity explained variation in dormant cladoceran assemblages. The analysis
of the dormant assemblages was essential, not only in revealing the potential species richness but also for better understanding
assemblage structure of aquatic organisms.

Key words: Bolivia, Andean temporary pools, Resting eggs, Bolivian zooplankton, Egg-morphotype.

Unidad de Limnología y Recursos Acuáticos (ULRA), Universidad Mayor de San Simón, Cochabamba, Bolivia. Tel: ++591 (4) 423 5622; E-mail: js.crnl@gmail.com
1

Laboratory of Aquatic Ecology and Evolutionary Biology, Katholieke Universiteit Leuven, Naamsestraat 59 – 3000 Leuven, Belgium.
2

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 REVISTA BOLIVIANA DE ECOLOGÍA Y CONSERVACIÓN AMBIENTAL
INTRODUCTION
The cyclical and variable nature of the temporary pool
environment creates a habitat that is quite distinct from
permanent and more predictable habitats such as ponds
and lakes. Their inhabitants require specific adaptations
to deal with the variable and often extreme local envi-
ronmental conditions, which often results in the presence
of unique species not found in any other habitat types
(Williams, 1997; Blaustein & Schwartz, 2001; De Meester
et al., 2005).
A main feature of aquatic species permanently inhabiting
variable aquatic environments is the production of dia-
pausing resting stages (dormant eggs) that allow survival
during droughts and recolonization after flooding (Wiggins
et al., 1980; Cáceres, 1997). Zooplanktonic organisms,
in particular, produce diapausing stages when environ-
mental conditions become adverse (Brendonck & De
Meester, 2003). After deposition, most resting stages
sink to the bottom building a dormant egg bank. The lar-
gest fractions of viable (responsive) eggs usually occur
in the top three centimeters, the so-called “active egg
bank” (Brendonck & De Meester, 2003). At each occa-
sion, usually only a variable portion of the egg bank hat-
ches (Maia-Barbosa et al., 2003; García-Roger et al.,
2006). Such partial hatching constitutes a bet-hedging
strategy to buffer against extinction in the variable habitat,
where sometimes there is even not enough time for ma-
turation and successful reproduction (Brendonck & De
Meester, 2003). The portion of resting eggs that does
not hatch at the first occasion may do so later under si-
milar conditions (Brendonck et al., 1998). This process
results in the gradual accumulation of dormant eggs
from different seasons and years in a persistent egg
bank, integrating spatial and temporal variation in the
abundance and distribution of freshwater zooplankton
(Brendonck & De Meester, 2003). This property renders
the dormant egg bank a potentially valuable tool for
detecting hidden diversity with only a limited presence
in the water column and an attractive complementary
tool for the analysis of active community samples (Vande-
kerkhove et al., 2005a; Vandekerkhove et al., 2005b).
The study of the dormant egg bank in permanent aquatic
systems revealed that the number of species hatching
from sediment samples in the laboratory was usually
higher than the number of species detected by snapshot
sampling from the active community (May, 1986; Vande-
kerkhove et al., 2005b). Some studies on temporary
pools, however, revealed a lower number of species
than detected in the active community samples (Boven
52
& Brendonck, 2008). Hatching of dormant eggs can be
biased because some populations seldom or never
produce dormant eggs (Jeppesen et al., 2003) or due
to variation among taxa in their response to specific
hatching stimuli (Cáceres, 1998).
The shape and external sculpturing (ornamentation) of
resting eggs were also suggested as a tool for identi-
fication at a higher taxonomic level and, in some cases,
even to species level (reviewed in Brendonck & De
Meester, 2003). Vandekerkhove et al., (2004a) showed
that morphological characters of resting eggs allowed
a rapid first estimation of cladoceran species richness
in 20 shallow European lakes.
In this study we tested the efficiency of using egg bank
samples for detecting potential species richness in
temporary peat land pools in the high Andes of Bolivia.
By individual incubation of isolated egg morphotypes
from 61 pools we specifically aimed to 1) assign ephippia
morphotypes to species, 2) obtain a more integrated
picture of peat land cladoceran species richness and
composition by comparing the active and dormant
cladoceran assemblages, and 3) explore whether the
same environmental variables explain variation in
structure of dormant compared to active communities.
MATERIAL AND METHODS
Study site
Our study area is located in the Cordillera del Tunari
(Cochabamba) in the Bolivian Andes between the coor-
dinates 66o08’- 66o22’ W and 17o10’-17o19’ S, at altitudes
ranging from 4000 to 4400 m.a.s.l. This area consists
of numerous small peat land systems (locally called bo-
fedales) scattered over valleys and mountain slopes.
Most of these peat lands contain small temporary peat
land pools, of which the number typically varies between
1 and 8, although pools can be more numerous in some
of the larger peat land systems (Fig. 1). Peat land pools
are mostly temporary and fishless, characterized by high
water transparency and low values of conductivity, sali-
nity, and total dissolved solids. The area is in general
cha-racterized by grassland vegetation with exception
of the peat land that is predominantly dominated by a
hard-tapestry vegetation of Distichia muscoides and
Plantago tubulosa (Navarro & Maldonado, 2002). This
area is subject to a dry (April – September) and rainy
(October – March) season.
 CORONEL, J.S, X., AGUILERA, S., DECLECK & L., BRENDONCK: Running head: Hidden Andean diversity
Figure 1. Bofedal showing high-altitude peatland pools in
the Cordillera del Tunari of the Bolivian Andes.
Sampling
Samples of both, the active zooplankton community and
the dormant egg bank, were collected in 61 peat land
pools from 31 peat land systems spread over four
mountain valleys in the Cordillera del Tunari: Taquiña,
Toro, Saito, and San Ignacio (Fig. 2).
The active community was collected in the middle of the
wet season between February and March 2004. In each
peat land pool, 3 to 15 L samples were taken with a tube
sampler (75 mm diameter and 1.5 m length) and filtered
through a 30-µm mesh. Samples were preserved in
sucrose-formaldehyde solution (5% final concentration).
In the dry season, for the dormant egg bank analysis,
the top three centimeters of sediment were collected
using a KC-sediment core sampler (0.7 meter long plexi-
glass tube of 5.2 cm diameter), until completing one
kilogram per pool. After collection, samples were wrapped
in aluminum foil and transported to the lab in a cooler
box.
For each peat land pool, the following environmental
variables were recorded: pH, conductivity (COND), chlo-
rophyll-a (CHLa), pool surface (AREA), and water column
depth (DEPTH). Besides, we also measured the thickness
of the bottom sediment (SEDTH, measured as the
thickness of the bottom organic matter layer), the per-
centage of water plants covering the pool (WPCOV),
connectivity (CONN; measured as the number of peat
land pools that directly drained into the sampled pool),
number of neighboring peat land pools in a radius of 50
m (NGP), and distance to the nearest rivulet (DNR).
53
Cordillera del Tunari
N 17o
Toro
Saito
San Ignacio
Taquiña
Cochabamba
20 Kilometers
18o
66o
65 o
Figure 2. Map of the study area. Stars indicate the study
mountain valleys in the Cordillera del Tunari in Cochabam-
ba, Bolivia.
Sample Analysis
Zooplankton density estimates were based on counts
of at least 300 specimens per sample. The density of
potential zooplankton predators (cyclopoid copepods,
mites, and larvae of the coleopteran genera Ranthus,
Colymbetinae and Hydroporus, Hydroporinae) was also
assessed by counting specimens in each sample. Zoo-
plankton and potential predators were counted using an
Olympus SZX12 stereo microscope.
In order to isolate dormant eggs of each individual peat
land pool we removed gross material (mostly vegetal
debris) from each sample using sieves of 1000 µm and
500 µm, while fine material and resting eggs were
retained on a 63 µm sieve.
The retained resting eggs were isolated by the sugar
flotation method (Onbé, 1978; Marcus, 1990). We omitted
the sonication step because none of the sediment
 REVISTA BOLIVIANA DE ECOLOGÍA Y CONSERVACIÓN AMBIENTAL
samples were compact. The following steps of the original
Onbé-Marcus method were retained: 1) filtration through
a 48 µm mesh, 2) centrifugation of the residue in a sugar
solution (1000 g table sugar in 1000 ml distilled water)
at 3000 rpm for three minutes, and 3) washing of the
supernatant over a 48 µm mesh using tap water.
The isolated resting eggs were sorted on the basis of
morphology and counted under a stereo microscope
(Olympus SZX12). Dormant egg density estimates were
based on counts of at least 300 specimens per sample.
To allow identification to species level, the unknown
resting egg types were incubated individually in 30-ml
multi-well plates containing the Aachener Daphnien
Medium (ADAM; Klüttgen et al., 1994) at a conductivity
of 30 µS cm-1. Multi-well plates were placed in an
incubator at 20oC with a photoperiod of 14 hours light
and 10 hours dark. Incubation medium was refreshed
every five days. For a period of two months, all multi-
well plates were checked every four days for emerging
hatchlings. Hatchlings were transferred to 50-ml vessels
and fed Scenedesmus obliquus (100.000 cells ml-1) until
maturation. All cladocerans were identified to species
level using Paggi (1995), Alonso (1996) and Smirnov
(1996).
Statistical analyses
Differences in composition between cladoceran assem-
blages obtained from both the dormant egg bank and
active cladoceran samples were tested with permutation
tests (999 permutations) on redundancy analysis models
(RDA; CANOCO 4.5). Nominal dummy variables cons-
tructed for the two cladoceran assemblages (dormant
and active) were used as explanatory variables and the
densities of individual species as response variables.
Cladoceran densities were square-root transformed to
minimize the effect of high densities (ter Braak & Šmilauer,
2002).
To test for differences in species richness between dor-
mant and active assemblages across all peat land pools
sampled (n = 61) we used a paired T-test for dependent
samples. In addition, the degree of association between
pool species richness derived from the dormant egg
bank analysis and species richness from active clado-
ceran samples was evaluated using product moment
correlations. Associations between environmental va-
riables and species richness of the dormant and active
assemblages were also evaluated with product moment
correlations using the statistical software STATISTICA
V8.0, Statsoft INC., Tulsa, O.K.
54
To explore for environmental variables that explain va-
riation in community structure of cladoceran assemblages
obtained from dormant versus active community samples,
we used a standardized redundancy analyses (RDA).
All recorded environmental variables (explanatory va-
riables) were log (X+1) transformed except for pH. The
RDA model was constructed using forward selection
(999 Monte Carlo permutations). Only significant variables
were retained. We evaluated the amount of variation
explained by each environmental variable included in
the model. Cladoceran densities (response variable)
were square-root transformed to minimize the effect of
high densities (ter Braak & Šmilauer, 2002). Only peat
land pools with samples allowing counts of at least 300
individuals were included in the analysis. Cladoceran
species with less than 5% of pool-occurrence were ex-
cluded from the analysis, since they can disproportionately
affect the results.
RESULTS
Ephippia morphotype analysis
We identified 24 different morphotypes of cladoceran
ephippia down to species level (Table 1; Appendix 1).
Most of the isolated ephippia contained one egg with
exception of those belonging to Daphnia pulex, Daphnia
peruviana, Macrothrix atahualpa, Ilyocryptus cf spinifer,
Paralona piagra and Streblocerrus serricaudatus that
presented two eggs. A similar morphotype was observed
for Ceriodaphnia cf dubia and C. cf laticaudata. On
average, most cladoceran ephippia hatched during the
first 8 days of incubation (Table 1).
Composition and species richness of dormant and
active assemblages
Composition of the cladoceran assemblages obtained
from the resting egg bank versus those from the active
cladoceran samples differed significantly (RDA analysis:
Trace= 0.123; F= 17.2; p= 0.001).
The study of the dormant egg bank yielded 24 cladoceran
species, in comparison with 21 species in the active
cladoceran samples (Fig. 3). The number of cladoceran
species retrieved from the resting egg bank was signi-
ficantly higher (mean per pool: 6.7 0.6) than the number
collected from the active assemblages (mean per pool:
4.6 0.4) (paired T-test = 6.3; df = 60; p = < 0.001; Fig.
4).
 CORONEL, J.S, X., AGUILERA, S., DECLECK & L., BRENDONCK: Running head: Hidden Andean diversity

Table 1. Cladoceran species detected in temporary peat land pools in the Cordillera del Tunari, Bolivia. Species are
ordered according to their frequency of occurrence (from high to low) in the 61 study peat land pools. ‘Average
densities’ refers to the mean number of eggs (eggs per Kg) for the dormant community and the mean number of
individuals (individuals per liter) for the active community, across all study pools. ‘DFH’ shows the number of days
for first hatch to occur. ‘Valley’ shows the mountain valleys where the respective species were found: a = Taquiña,
b = Toro, c = Saito, d = San Ignacio.

Species’ occurrence Average densities DFH Valley

Dormant Active Dormant Active

Chydorus brevilabris 52 59 26.30 71.63 6 abcd
Alona ossiani 55 51 48.67 6.47 8 abcd
Macrothrix atahualpa 53 46 61.75 10.40 6 abcd
Alona cambouei 42 44 26.20 3.04 6 abcd
Simocephalus mixtus 47 37 35.20 6.30 7 abcd
Alonella excisa 30 48 8.49 5.04 6 abcd
Camptocercus aloniceps 23 24 9.26 2.07 10 abcd
Alona davidi 17 12 9.85 0.68 6 abcd
Ceriodaphnia cf dubia 22 14 9.36 2.40 6 bc
Graptoleberis testudinaria 11 9 3.67 0.30 5 abc
Daphnia pulex 7 6 1.31 0.07 9 bcd
Alona glabra 4 6 0.74 0.05 10 abd
Daphnia peruviana 5 5 0.16 0.17 9 bc
Ephemerophorus hibridus 7 2 4.28 0.30 10 abc
Paralona piagra 6 3 0.67 1.54 15 a
Pleuroxus caca 0 9 0.00 0.17 10 bcd
Ceriodaphnia cf laticaudata 8 0 3.34 0.00 ab
Pleuroxus sp. 6 0 1.38 0.00 7 abcd
Streblocerus serricaudatus 5 0 2.92 0.00 10 a
Alona boliviana 3 1 0.75 1.02 7 ab
Scapholeberis spinifera 2 1 0.08 0.02 11 ac
Drepanothrix cf dentata 2 0 0.44 0.00 8 ac
Pleuroxus cf aduncus 2 0 0.13 0.00 ac
Alona.cf ossiani 1 0 0.30 0.00 b
Bosmina huaronensis 0 1 0.00 0.00 a
Ephemerophorus cf acanthodes 0 1 0.00 0.01 c
Ilyocryptus cf spinifer 1 0 0.11 0.00 16 b
Pleuroxus hardingi 0 1 0.00 0.10 b

Seventeen cladoceran species were detected in both
the active and dormant assemblages (Table 1), whereas
seven species (Ceriodaphnia cf laticaudata, Pleuroxus
cf aduncus, Pleuroxus sp., Streblocerus serricaudatus,
Drepanothrix cf dentata, Alona cf ossiani, and Ilyocryptus
cf spinifer) were exclusively observed in the dormant
assemblage (Table 1). Some cladoceran species were
uniquely present in the active cladoceran samples
(Pleuroxus caca, Bosmina huaronensis, Ephemerophorus
cf acanthodes, Pleuroxus hardingi) (Table 1). The accu-
mulated (active plus dormant) cladoceran species rich-
ness of the study pools mounted up to 28 species (Table
1).
Species richness of active cladoceran samples was
significantly associated with water plant coverage and
with species richness of the dormant community (Table
2).
Explanatory environmental variables of dormant and
active assemblages
For the active assemblage, the forward selection
procedure of the RDA analysis indicated water plants
as the main environmental variable explaining variation
in community structure (Trace = 0.063; F = 2.07; p =
0.49; Table 3).
For the dormant egg banks, a model including sediment
thickness, connectivity and chlorophyll-a explained a
significant portion of the variation in the assemblage
structure (Trace = 0.182; F = 2.15; p = 0.002; Table 3).

55
 REVISTA BOLIVIANA DE ECOLOGÍA Y CONSERVACIÓN AMBIENTAL

26
I. cf spinifer and S. spinifera) were easier to identify than
24 others because they still maintained some ornamental
22 characters of active individuals. Adults of Alona glabra,
20 for instance, usually present a tuberculated ornamentation
in their shield. This ornamentation was maintained in
Species richness

18

16
their ephippia (Appendix 1). Cladoceran identification
solely based on ephippia morphotypes may result in an
14
underestimation of true species richness as similar
12
morphotypes sometimes occur among different species
10
(Vandekerkhove et al., 2004a). In our study, morpholo-
8 gically similar ephippia were observed in species within
6 the genera Ceriodaphnia and Pleuroxus.
4
0 10 20 30 40 50 60 70
Although cladoceran assemblage composition derived
Number of pools from the resting egg bank resembled the assemblage
obtained from active samples, on average 45% more
Figure 3. Species accumulation curves for cladoceran cladoceran species per peat land pool were detected
assemblages derived from active (empty symbols) and
by egg bank analysis. This discrepancy may actually be
dormant (filled symbols) samples across 61 high-altitude
temporary peat land pools of the Cordillera del Tunari, even much higher as still some species may not have
Bolivia. hatched at all (bet-hedging effects) in the single incubation
event (no multiple inundations). To our knowledge, a
higher species yield from dormant egg banks versus
14 ac-tive assemblage samples has mostly been reported
for permanent lakes (May, 1986; Havel et al., 2000;
Cladocera species richness

12 Crispim & Watanabe, 2001; Vandekerkhove et al., 2004a;

2005a; 2005b). Vandekerkhove et al., (2004b) found on
10
average 35% more cladoceran species per lake in
8 dormant egg banks of 95 European permanent lakes
than in the corresponding active assemblage samples.
6 In rotifers from lake Loch, Great Britain, the assemblage
4
of resting eggs also contained higher species richness
than in the water column in any single year, but was
2 fully concordant with the assemblage observed over a
six year period (May, 1986).
0

-2 Although our results underline the efficiency of resting

ACS
Figure 4. Average of the total number of cladoceran species
recovered per pool from samples of the dormant egg bank
(DEB) and from snapshot active samples (ACS). Errors
bars represent 1 standard error of the mean.
DISCUSSION
The observed number of egg morphotypes in the dormant
egg bank yielded higher number of cladoceran species
than in active samples. Incubation of isolated unknown
egg morphotypes allowed identifying cladoceran ephippia
down to species level (Appendix 1). Some cladoceran
ephippia (e.g. A. glabra, C. aloniceps, G. testudinaria,
DEB egg bank analysis for species richness assessment of
zooplankton assemblages in temporary peat land pools,
other studies on temporary aquatic systems revealed
opposite results. In Kiskunság National Park (Hungary),
for instance, the resting egg banks of 12 temporary
pools yielded only 19 cladoceran species out of a total
of 32 species observed in the active assemblage samples
(Boven & Brendonck 2008). Incubation of the resting
egg banks from eight temporary pools in South Africa
did not yield higher species richness than active assem-
blage samples (De Roeck et al., in press). In these
examples, hatching success and low species yield may
have resulted from bet-hedging effects which are expec-
ted to be more important in the variable environment of
temporary pools. Bet-hedging occurs when only a fraction
of all viable resting eggs hatches under ideal conditions

56
 CORONEL, J.S, X., AGUILERA, S., DECLECK & L., BRENDONCK: Running head: Hidden Andean diversity

Table 2. Pearson correlations for environmental variables and species richness of dormant and active community
samples. Only peat land pools with samples allowing counts of at least 300 individuals were included in the analysis.
Codes: TP = total phosphates, TN = total nitrates, pH = pH, COND = conductivity, ALK = alkalinity, TRANSP = water
transparency (Snell measure), DEPTH = water column depth, AREA = pool surface area, WPCOV = water plant cover,
CHLa = chlorophyll a, DNR = distance to the nearest rivulet, NGP = neighbor pools, CONN = connectivity, SEDTH =
thickness of the bottom sediment, Mac. Pred. = macroinvertebrate predators, Act. Sp. Rich = species richness from
active communities, Dor. Sp. Rich = species richness from dormant communities. **: P<0.01; ***: P<0.001.

TP TN pH COND ALK TRANSP DEPTH AREA WPCOV CHLA DNR NGP CONN SEDTH Mac. Act.
Pred. Sp.
Rich

TP
TN 0.10
pH 0.16 0.17
COND 0.04 0.07 0.24
ALK 0.18 0.05 0.24 0.31
TRANSP 0.08 0.09 -0.21 -0.26 0.02
DEPTH 0.14 0.52** 0.03 0.00 0.12 0.52**
AREA 0.14 0.08 0.25 -0.08 -0.24 0.17 0.26
WPCOV -0.46** -0.03 -0.23 0.08 -0.30 -0.21 -0.10 0.12
CHLa -0.13 -0.25 0.26 0.09 -0.07 -0.58*** -0.31 0.01 0.13
DNR 0.14 -0.02 -0.07 -0.08 -0.05 0.01 0.03 0.30 0.26 0.01
NGP 0.26 0.23 0.10 -0.15 0.30 0.01 0.04 -0.03 -0.04 0.06 -0.02
CONN -0.31 0.15 0.18 -0.03 0.08 -0.14 -0.01 0.02 -0.08 0.00 -0.14 0.31
SEDTH -0.06 0.29 -0.28 -0.16 0.13 0.18 0.26 0.06 0.06 -0.33 -0.04 0.13 -0.09
Mac. Pred. 0.12 0.11 -0.07 -0.13 -0.23 -0.31 -0.06 0.28 0.05 0.07 0.21 -0.17 0.08 0.07
Act. Sp. 0.29 -0.11 0.18 -0.11 0.08 0.14 0.17 0.31 -0.46** 0.09 0.12 -0.02 -0.12 -0.20 0.08
Rich
Dor. Sp. 0.33 -0.11 0.20 0.16 0.15 0.14 0.24 0.14 -0.28 0.24 0.10 0.05 -0.28 -0.33 -0.17 0.73***
Rich

Table 3. Environmental variable models explaining variation in cladoceran assemblages in dormant and active samples.
The total variation explained by the model, the percentage contributions of its constituents, and significance values
are shown. Only peat land pools with samples allowing counts of at least 300 individuals were included in the analysis.
See materials and methods for an explanation of the variable codes.

Total %
Variables Co-variables F-ratio p-value
Variation Variation

Active community samples

Entire model

WPCOV 0.063 2.079 0.049

Dormant community samples

Entire model

SEDTH, CHLa, CONN 0.182 2.156 0.002

Individual contribution
SEDTH CHLa, CONN 0.089 48.9 3.147 0.002
CHLa SEDTH, CONN 0.069 37.9 2.445 0.020
CONN SEDTH, CHLa 0.054 29.6 1.928 0.046

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 REVISTA BOLIVIANA DE ECOLOGÍA Y CONSERVACIÓN AMBIENTAL
(Brendonck et al., 1998; Brendonck & De Meester, 2003).
In our study, the detection of cladoceran species living
in close association with the substrate (S. serricaudatus,
D. cf dentata, and I. cf spinifer) and that were therefore
absent from active assemblage samples shows the
effectiveness of the dormant egg bank analysis (Dole-
Olivier et al., 2000; Tremel et al., 2000; Fefilova et al.,
2006). Similar observations were reported by Vande-
kekhove et al., (2005b) who collected benthic taxa like
Ilyiocryptus sp, Alona sp., Leydigia sp., and Pleuroxus
sp., from the dormant egg bank. In our study, the absence
of the above mentioned species in the active samples
may be influenced by seasonal dynamics. Two months
of field sampling was too limited to collect cladoceran
species occurring at different times during the inundation
cycle. Seasonal dynamics in invertebrate communities
was shown as a typical phenomenon, even in temporary
pools (Lake et al., 1989; Lahr et al., 1999; Jocqué et al.,
2007; Boven & Brendonck, 2009).
Cladoceran species only observed in the active assem-
blage (P. caca, P. hardingi, B. huaronensis, and E. cf
acanthodes) were probably missed from dormant
samples due to their low densities in the egg banks (see
Table 1). Moreover, these species occurred in very low
densities in the active assemblage in a specific mountain
valley (Table 1).
Different environmental variables explained variation in
the cladoceran assemblages of the active and dormant
samples (Table 3). Water plants were more important
for the active assemblages (Table 3). Species richness
was inversely correlated with water plant density
(r =-0.46, p = 0.001; Fig. 5a). Although vegetation may
increase the number of habitats, niches and food
resources and reduce the susceptibility of aquatic inver-
tebrates to fish predation (Jeppesen et al., 1998; Diehl
& Kornijów, 1998), in fishless systems, water plants may
increase invertebrate predation pressure rather than
being a shelter (Meerhoff et al., 2006; 2007).
Dormant egg bank analysis revealed important environ-
mental variables that structure zooplankton assemblages.
In our study, chlorophyll-a was an important variable
structuring dormant assemblages. This result shows the
effectiveness of the dormant egg bank in detecting
environmental variables which cannot be detected by
the analysis of active assemblages. A data set with an
increased number of species probably offers a better
resolution in the analyses. Although a correlation does
not imply a cause – effect, there was a positive correlation
58
between species richness of the dormant egg bank and
chlorophyll-a (Fig. 5b).
The thickness of the sediment layer may influence
hatching requirements. Resting eggs displayed over thin
bottom sediments are certainly younger than those ones
buried in thick bottom sediments. Younger eggs rapidly
respond to environmental stimuli resulting in high rates
of hatchlings (Brendonck & De Meester, 2003). Sediment
layer thickness was negatively correlated with species
richness of the dormant assemblage (r =-0.33, p = 0.051;
Fig. 5c).
Connectivity significantly explained variation in dormant
assemblages but was not included in the explanatory
model for the active ones (Table 3). Species richness
decreased with connectivity (Fig. 5d). It is likely that co-
mmunity homogenization occurs in more connected peat
land pools through dispersal of dormant eggs. Patterns
of higher species richness in pools of intermediate iso-
lation in comparison with highly connected pools were
also revealed by Vanschoenwinkel et al., (2007) in
temporary rock pools in South Africa. It remains unclear
why this pattern was revealed in our study only in dormant
and not in active communities. Probably the pattern was
obscured in active communities as several species were
missed. Further studies should elucidate whether these
species mainly occurred in isolated systems.
Our results suggest that the selection of resting egg on
the basis of morphotypes and their subsequent hatching
is a reliable method to estimate cladoceran species
richness in temporary high-altitude peat land pools. This
method may replace labor intensive field sampling, par-
ticularly in areas of difficult access and harsh climatic
conditions. However, care must be taken since different
species can produce morphologically similar eggs.
Results of assemblage structure analyses revealed that
variables explaining variation in dormant assemblages
were not concordant with those explaining variation in
active ones. This may result from the fact that egg banks
integrate eggs produced during years of variable environ-
mental conditions, while the analysis was done on the
basis of single moment measurements. On the other
hand, higher species yields by incubation of the egg
bank may have contributed to a higher resolution to re-
veal assemblage structuring patterns. Analysis of the
dormant egg bank is therefore invaluable, not only in re-
vealing the potential species richness but also for better
understanding assemblage structure of aquatic orga-
nisms.
 CORONEL, J.S, X., AGUILERA, S., DECLECK & L., BRENDONCK: Running head: Hidden Andean diversity

a 1,20
b
1,15
1,15
1,10
1,10

Species richness DEB

1,05
Species richness ACS

1,00 1,05

0,95 1,00
0,90
0,95
0,85
0,90
0,50
0,85
0,75
0,80
0,70

0,65 0,75
1,55 1,60 1,65 1,70 1,75 1,80 1,85 1,90 1,95 2,00 2,05 0,2 0,4 0,6 0,8 1,0 1,2 1,4 1,6 1,8 2,0 2,2 2,4
Water plant coverage Chlorophylla
1,20 c
1,20 d
1,15
1,15
1,10
1,10
Species richness DEB

Species richness DEB

1,05
1,05
1,00
1,00

0,95
0,95

0,90 0,90

0,85 0,85

0,50 0,80

0,75 0,75
0,4 0,6 0,8 1,0 1,2 1,4 1,6 1,8 2,0 -0,2 0,0 0,2 0,4 0,6 0,8 1,0 1,2
Thickness of the bottom sediment Connectivity

Figure 5. Scatter plots showing the associations between ecological relevant variables for species richness recovered
from the dormant egg bank (DEB) and from active samples (ACS). Correlations were based on peat land pools with
samples allowing counts of at least 300 individuals.

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Appendix 1: Resting eggs morphotypes detected in the present study. Each picture shows the name of the species
hatched from a given morphotype. Characteristics: number of eggs per ephippium, size (length and width) and other
morphological characteristics used in the present study to identify morphotypes.

Characteristics:

1 egg
Length: 156µm, width: 90µm
Narrowing at posteroventral side, brown-dark color
Very alike to A. ossiani

Alona boliviana 100 µm

Characteristics:

1 egg
Length: 205µm, width: 153µm
Two detectable ridges on the egg

Alona cambouei 100 µm

Characteristics:

1 egg
Length: 113µm, width: 84µm
Transparent color
Egg covered by oval-shape valves
Very alike to A. ossiani

Alona cf ossiani 100 µm

Characteristics:

1 egg
Length: 229µm, width: 122µm
Narrowing at posteroventral side
Longitudinal ridges are observed.

Alona davidi 100 µm

62
 CORONEL, J.S, X., AGUILERA, S., DECLECK & L., BRENDONCK: Running head: Hidden Andean diversity

Cont. Appendix 1.

Characteristics:

1 egg
Length: 164µm, width: 105µm
Oval shape
Tuberculated ornamentation.

Alona glabra 100 µm

Characteristics:

1 egg
Length: 135µm, wide: 79µm
Narrowing at posteroventral side
Without sculpturing on the valves
Very alike to A. boliviana.

Alona ossiani 100 µm

Characteristics:

1 egg
Length: 162µm, width: 103µm
Oval shape
Valves with a net like ornamentation

Alona excisa 100 µm

Characteristics:

1 egg
Length: 127µm, width: 67µm
Oval shape
Longitudinal ridges on the valves

Camptocercus aloniceps 100 µm

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 REVISTA BOLIVIANA DE ECOLOGÍA Y CONSERVACIÓN AMBIENTAL

Cont. Appendix 1.

Characteristics:

1 egg
Length: 95µm, width: 67µm
Oval shape
Often with floating cells

Ceriodaphnia cf dubia 100 µm

Characteristics:

1 egg
Length: 152µm, width: 135µm
* Like circular shape
Prominent postero ventral angle (*)

Chydorus brevilabris 100 µm

Characteristics:

2 eggs
Length: 167µm, width: 123µm
Egg in the dorsal part
Darkly colored

Daphnia peruviana 100 µm

Characteristics:

2 eggs
Length: 191µm, width: 119µm
Egg in the dorsal part
Dark-brown colored

Daphnia pulex 100 µm

64
 CORONEL, J.S, X., AGUILERA, S., DECLECK & L., BRENDONCK: Running head: Hidden Andean diversity

Cont. Appendix 1.

Characteristics:

2 eggs
Length: 93µm, width: 88µm
Egg in the dorsal part
Transparent

Drephanotrix cf dentata 100 µm

Characteristics:

1 eggs
Length: 124µm, width: 109µm
Egg in the dorsal part, circular shape
Brown color

Ephemerophorus sp 100 µm

Characteristics:

1 eggs
Length: 239µm, width: 141µm
Egg in the dorsal part, oval shape
Brown colored with hexagons-like sculpturing on the valves

Graptoleberis testudinaria 100 µm

Characteristics:

2 eggs
Length: 109µm, width: 103µm
Transparent egg and circular shape
Like tuberculated valves with concentric rings

Ilyocryptus cf spinifer 100 µm

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 REVISTA BOLIVIANA DE ECOLOGÍA Y CONSERVACIÓN AMBIENTAL

Cont. Appendix 1.

Characteristics:

2 eggs
Length: 101µm, width: 87µm
Egg in the dorsal part
Brown-dark colored

Macrothrix atahualpa 100 µm

Characteristics:

2 eggs
Length: 146µm, width: 130µm
Curved ridges on the valves
Transparent egg

Paralona piagra 100 µm

Characteristics:

1 egg
Length: 161µm, width: 126µm
Brown-dark colored

Pleuroxus aduncus 100 µm

Characteristics:

1 egg
Length: 171µm, width: 171µm
Prominent ridges and depressions on the valves
Brown-dark colored

Pleuroxus caca 100 µm

66
 CORONEL, J.S, X., AGUILERA, S., DECLECK & L., BRENDONCK: Running head: Hidden Andean diversity

Cont. Appendix 1.

Characteristics:

1 egg
Length: 177µm, width: 139µm
Rhombus-like sculpturing on the valves
Brown-dark colored

Pleuroxus sp 100 µm

Characteristics:

1 egg
Length: 94µm, width: 61µm
No sculpturing on the valves
Intense brown-dark colored

Schapholeberis spinifera 100 µm

Characteristics:

1 egg
Length: 177µm width: 107µm
Narrowing sharply at posteroventral side
No sculpturing on the valves with floating cells
Intense brown-dark colored

Simocephalus mixtus 100 µm

Characteristics:

2 eggs
Length: 161µm, width: 153µm
Small rhombus-like sculpturing on the valves
Brown-dark colored

Streblocerus serricaudatus 100 µm

67