Primary Research Paper

Local and regional factors determining aquatic and semi-aquatic bug

(Heteroptera) assemblages in rivers and streams of Greece
Ioannis Karaouzas* & Konstantinos C. Gritzalis
Hellenic Centre for Marine Research, Institute of Inland Waters, 46.7 km Athens-Sounion Av., P.O. BOX 712, 190 13,
Anavissos, Attica, Greece
(*Author for correspondence: Tel.: +30-22910-76391; Fax: +30-22910-76323; E-mail: ikarz@ath.hcmr.gr)
Received 21 December 2005; in revised form 15 June 2006; accepted 17 June 2006; Published online 31 August 2006
Key words: Heteroptera, assemblages, distribution, variation partitioning, multivariate analysis, environmental fac-
tors, Greece
Abstract
Heteroptera species were collected from48 sites distributed throughout the mainland and island complexes of
Greece during 19992004. The aims of this study were to investigate Heteroptera distribution and abundance
in Greek streams, identify the environmental factors that are linked to variation in their assemblages and to
partition the inuence of environmental and spatial components, alone and in combination, on Heteroptera
community composition. Canonical ordination techniques (CCA) were used to determine the relationship
between environmental variables and species abundance, while variation partitioning was performed using
partial CCA to understand the importance of dierent explanatory variables in Heteroptera variation.
Heteroptera variation was decomposed into independent and joint eects of local (physicochemical variables,
microhabitat composition, stream width and depth), regional (land use/cover) and geographic variables
(longitude, latitude, altitude and distance to source). Land use/cover, aquatic and riparian vegetation, stream
size and water chemistry were the most important factors structuring Heteroptera assemblages. At regional
scale, bug assemblages were mainly divided into those found in forested and agricultural landscapes,
following water quality andmicrohabitat composition at local scale. Local variables accounted for 48%of the
total explained variation, regional variables for 20% whereas geographical position appeared to be the least
inuencing factor (8.5%). The results of partial constraint analyses suggested that local variables play a major
role in Heteroptera variation followed by regional variables.
Introduction
Aquatic Heteroptera, commonly known as true
bugs, are an important group of aquatic insects that
possess unique characteristics and adaptations.
The most distinguished characteristic of this order
is the beak, a piercing mouthpart, which has a
suctorial function. Each family of this order diers
considerably in morphology and ecological pref-
erences while many species display specic habitat
preferences (i.e., Corixidae) (Macan, 1954; Savage,
1990, 1994a). In rivers, they are found along the
margins of shallow water (Corixidae), on the water
surface of lentic (pool) stream zones (Gerridae
and Veliidae) and lotic (rie) stream zones
(some Veliidae), and among aquatic vegetation
(Notonectidae, Nepidae and Naucoridae). They
may also be found under rocks in fast waters (some
Naucoridae). Aquatic Heteroptera have signicant
ecological eects (McCaerty, 1981; Hutchinson,
1993) and economic importance, which has
probably been underestimated (Papacek, 2001).
Hydrobiologia (2006) 573:199212 Springer 2006
DOI 10.1007/s10750-006-0274-1
Ecologists studying Heteroptera have long been
seekingthe use of freshwater bugs for biomonitoring
and classication purposes. Prior to the develop-
ment of monitoring models, many studies have
been conducted in the past to ascertain bug species
association with environmental parameters. Bug
species associations were rst examined by Macan
(1938) who reported the relationship of Corixid
species with marginal vegetation and water
chemistry. Macan (1939) also provided general
ecological notes on each species of the Corixidae
family in his identication key. Members of the
Society of British Entomology and other
researchers later on provided much information on
the ecological and geographical distribution of
Heteroptera species (Brown, 1948; Popham, 1949,
1950) that have been summarised in Savages key
(1989). Savage (1982, 1990) found close correlation
of bug species composition with water chemistry
followed by Eyre & Foster (1989) who found that
water acidity aects species composition. Savage
(1994b) used Corixid species as indicators of
organic pollution that was supplemented by the
saprobic values of Sla decek and Sla deckova (1994).
Jansson (1977, 1987) used Micronecta species as
indicators of water quality in Finish Lakes.
Recently, Hufnagel et al. (1999) proposed a new
approach for habitat characterisation obtained by
new indices and cenological values of bug species.
Distributional patterns and assemblages of this
insect group are well established in several coun-
tries around the globe (Savage, 1990; Moreno
et al., 1997; Biro, 2003), while their importance as
biological indicators for classifying water bodies,
especially in the United Kingdom, has been
established more than two decades (Savage, 1982).
Unfortunately, Heteroptera, among other insect
groups, have been rarely studied in Greece and
most of the few studies that exist have been
conducted by foreign scientists that collected
species from various geographic parts during short
periods of the year (Drosopoulos, 1980). These
studies however, involved only species records and
their geographical area of collection (e.g., Josifov,
1959; Illies, 1978; Magnien, 2000). Drosopoulos
(1980) was the rst to assemble records from
museums and literature, including his personal
collections, to complete a check list of both
terrestrial and aquatic bugs of Greece. Recently,
Petrakis & Roussis (2001) have examined the
bioindication value of Hellenic aquatic Heterop-
tera using an algorithmic approach. In addition,
they have also provided a check list of Greek
Heteroptera species with their distribution in seven
areas that they examined.
This work contributes to the general knowledge
of Greek river Heteroptera with the provision of
species records and their localities that will
supplement the Hellenic check lists provided by the
aforementioned authors. The objectives of this
work are (a) to investigate Heteroptera distribution
patterns in running waters, (b) to assess the envi-
ronmental factors that might explain their assem-
blage structure and (c) to partition the inuence of
environmental and spatial components, alone and
in combination, on Heteroptera variation.
Materials and methods
Site description
Greece is a small Mediterranean country with
an area of approximately 132,000 km
2
and is
divided into the mainland and surrounding island
complexes. Its recent geological morphology has
formed a multitude of basins, drained mainly by
small and medium sized rivers (Skoulikidis et al.,
2006). Running waters in the entire country range
from small streams in the highlands and in semi-
arid grasslands and islands, to medium and large
rivers and from ephemeral streams to perennial
rivers.
Heteroptera sampling
Heteroptera species were collected from 1999 to
2004 within the framework of two EU R&D
programs, AQEM (The Development and Testing
of an Integrated Assessment System for the Eco-
logical Quality of Streams and Rivers throughout
Europe using Benthic Macroinvertebrates) and
STAR (Standardisation of River Classications):
Framework method for calibrating dierent
biological survey results against ecological quality
classications to be developed for the Water
Framework Directive (2000/60/EC). The
AQEM-STAR (AQEM Consortium, 2002)
sampling method was used for the collection of
Heteroptera species within the two projects, while
200
within the STAR project the RIVPACS (Armitage
et al., 1983) method was additionally used.
The AQEM-STAR methodology is based on a
multi-habitat scheme designed for sampling major
habitats proportionally according to their presence
within a sampling reach. A sample consists of 20
replicates taken from all microhabitat types at the
sampling site with a share of at least 5% coverage,
which must be distributed according to the share of
microhabitats. Benthic macroinvertebrates were
collected using a rectangular hand net of
0.25 m0.25 m with a mesh size of 500 lm nytex
screen. Each of the 20 replicates was taken by
positioning the net and disturbing the substrate in
an area that equals the square of the frame width
upstream of the net (0.25 0.25 m). Thus, a total
of 1.25 m
2
(0.25 0.25 20 replicates) was sam-
pled for each sampling site. Sampling started at the
downstream end of the reach and proceeded up-
stream. Benthic samples were preserved with eth-
anol concentration of ca. 70%. Heteroptera species
were collected with soft tweezers from the samples
and identied with Savages (1989) and Tamaninis
(1979) keys.
The RIVPACS sampling method involves a
3-min kick sample and a 1-min manual search
which involves the collection of individual speci-
mens from the water surface, submerged rocks, logs
and vegetation. Each macroinvertebrate habitat in
the sampling area was sampled proportionally to its
cover with a pond net with a mesh size of 1 mm and
with a frame of 0.25 m0.25 m. Storage, preser-
vation as well as identication procedures were the
same to those of the AQEM-STAR method.
Environmental data
Prior to the collection of Heteroptera, the hydro-
morphology, land use/cover, geology of the site and
its catchment area, in-stream habitat composition
and physicochemical variables were examined and
recorded. Geographical, geological, morphological
and land use/cover data have been used in ArcView
GIS software to derive the respective information.
Particularly, land use data have been acquired by
using the CORINE Land Cover database of
Greece. The extent of the land cover classes at each
sampling catchment has been estimated with the
contribution of the ArcGIS Spatial Analyst, while
the surface areas have been rounded up to a 10%
spatial step. For the geological information a
preexisting geological map from the Institute of
Geologic and Mineralogic Exploration (IGME)
has been used in combination with the relevant
geologic formations database, which provided the
type of rocks and sediments in the area of interest.
Additionally, the altitude of each sampling station
has been estimated by utilizing the digital contour
map of Greece with an altitude step of 20 m
in ArcView. Physical parameters (estimated
discharge, water temperature, pH, conductivity
and dissolved oxygen) were measured in situ. Water
samples were collected, preserved at 4 C, ltered
upon arrival in the laboratory and analysed for
alkalinity, chloride, total hardness and nutrients
(nitrite, nitrate, ammonia, phosphate and total
phosphorous). In total, 40 environmental variables
were recorded.
Data analysis
Eighty stream sites were sampled in total, some
during three seasonal periods (summer, winter and
spring), during two (the vast majority of the sites)
or during one. Thus, for multiple seasonally
assessed sites the mean number of species abun-
dance was calculated. The same was conducted for
environmental data recorded more than one
season. Species collected with RIVPACS method
and were not found with the AQEM-STAR
method within a site, were included into the data
set in order to gain the highest possible number of
dierent bug species.
To nd groups of Heteroptera species based on
their occurrences, a hierarchical cluster analysis
was performed. The BrayCurtis similarity (pres-
enceabsence) and group average linkage were
used as similarity distance and linkage rule,
respectively. Canonical ordination techniques were
used to examine the relationship between the
environmental variables and species occurrence.
Prior to Canonical Correspondence Analysis
(CCA), variation in Heteroptera data was
examined by running a Detrended Correspondence
Analysis (DCA; Hill, 1979) to ensure a unimodal
rather than linear distribution. Gradient of varia-
tion is provided by the rst DCA axis in which
taxon compositional turnover is measured in
standard deviation units (SD). Along each axis a
full turnover in taxon composition between
201
samples occurs after 4.0 SD. The unimodal
assumption of DCA is accepted if the gradient
length of the rst axis is greater than 3.0 SD (Ter
Braak & Prentice, 1988). The rst DCA axis (SD:
4.576) conrmed the unimodal assumption and
thus the CCA application while the rst four DCA
axes accounted for 38.5% of the variation in Het-
eroptera data. To avoid multicollinearity between
environmental variables, a Pearson Product
Moment correlation analysis was run and those
variables highly associated with any other were
removed from the analysis, as they would have no
unique contribution to the regression equation (Ter
Braak, 1986). Forward selection of environmental
variables was used to ascertain the minimal set of
variables that explain species data. Signicance of
environmental variables was determined by means
of a Monte Carlo permutation test.
Partial canonical correspondence analysis
(CCA) (Borcard et al., 1992) was performed to
partition the variation of Heteroptera species into
independent components following the procedure
carried out by Qinghong (1997). Qinghong (1997)
used partial redundancy analysis (RDA) to parti-
tion the variation of species in his study. As bug
species in this study showed a unimodal rather than
a linear distribution, partial CCA was used instead.
Environmental variables were separated into three
groups of environmental data; geographic
(latitude, longitude, altitude and distance from
source), local (physicochemical variables, micro-
habitat composition, stream depth and width) and
regional (land use and cover) to partition the
inuence of each environmental group on the total
variance of bug species. Variation partitioning was
performed in two steps: (1) by running a partial
CCA of Heteroptera abundance (response)
variables and all three groups of environmental
variables (explanatory variables) and (2) by
running a partial CCA using one of the three
environmental groups (explanatory variables) and
the remainder two groups together (covariables)
and the other way around (Table 3, combinations
1, 2, 3), with or without the covariables (see
Table 3). Partial CCA was applied four times
within each combination of the three environ-
mental variable groups (see Table 3). A total of 12
runs were accomplished, in which the association
between species and an explanatory variable is
examined after the inuence of a covariable has
been separated. In this way, the pure inuence from
each environmental group and of the joint eects
was partitioned from the total explained variation.
Canonical ordination techniques were carried out
using the package CANOCO for Windows 4.5
(Ter Braak & Smilauer, 2002), correlation analysis
with STATISTICA version 6 and cluster analysis
with PRIMER 5.
Results
Heteroptera assemblages
From the 80 sampling sites, Heteroptera species
were found at 48 stream sites (Fig. 1). A total of 23
species (Table 1) were collected and identied
accounting for 424 individuals. It should be
mentioned that four species (Hebrus sp., Notonecta
sp., Velia sp., two individuals) at four sites, poor
specimen condition prevented identication of
Heteropterans and hence were removed from the
analysis.
Aquarius najas was the most abundant and
common (in terms of the number of sites which it
was found) species accounting for 28% of the total
species abundance and was found in 28 sites. Velia
caprai was the next most abundant species
accounting for 13% followed by Gerris lacustris
(9%), Notonecta maculata (8%) and Micronecta
poweri (6.5%). Following Aquarius najas, Velia
caprai, was the most widely distributed species
being found in 15 sites. Notonecta maculata, Gerris
lacustris and Hydrometra stagnorum were found in
14, 11 and 10 sites, respectively. At family level,
Gerridae, followed by Veliidae, were the most
abundant and common taxa, thus showing a
tolerance to a wide range of environmental fea-
tures. Notonectidae was the next most frequently
occurring family whereas Corixidae was the next
most abundant family. The highest species richness
was found in moderate to highly nutrient enriched
sites with pool (non-visible ow) waters and close
to the sea, in which 10 dierent species were found
in Kalipeuki and seven in Messini.
Cluster analysis revealed six groups of
Heteroptera assemblages (Fig. 2) based on
the strongest rankings of the dendrogram (less
than 20%). Starting from the right end of the
dedrogram towards the left, Plea minutissima and
202
the Corixids Sigara striata, Sigara falleni and
Sigara dorsalis formed group 1. Notonecta glauca
and Ilyocoris cimicoides formed a distinctive
group (group 2) while Velia caprai, Gerris
lacustris, Hydrometra stagnorum and Aquarius
najas formed group 3. Micronecta poweri and
Micronecta scholtzi comprised group 4. Velia
pelagonensis and Microvelia pygmaea created
group 5, which cannot however be considered as
valid, since a single individual of each species was
found and were commonly present in the same
stream site. The same also applies for Sigara
nigrolineata and Microvelia reticulata that formed
group 6.
Ordination analysis
Pearson ProductMoment Correlation analysis
revealed strong associations between various
environmental variables. For example, altitude
and slope (r = 0.90, p 0.01), conductivity and
total hardness (r = 0.80, p 0.01), alkalinity and
conductivity (0.83, p 0.01) and nally, nitrite
with ammonium, phosphate and nitrate (0.72, 0.71
Figure 1. Map showing the stream sites where Heteroptera species were found. Numbered below are the sampling sites followed by
their stream names. The small map on the upper right corner indicates the eco-region numbers (Illies, 1967) and the three hydro-
chemical zones of Greece (Skoulikidis et al., 2004). 1. Perasmata, Fonias; 2. Gria Vathra, Tsivdogianni; 3. Mesohori, Bospos; 4.
Symvola, Bospos; 5. Gorgona, Xanthia; 6. Ag.Barbara, Arkoudorema; 7. Dipotama, Arkoudorema; 8. Prasinada, Tributary of
Arkoudorema; 9. Thermia, Diavolorema; 10. Ano Poroia, Poroia; 11. Tripotamon, Lygkos; 12. Pidoderi, Aliakmon; 13. Adartiko,
Aliakmon; 14. Melas, Aliakmon; 15. Kotas, Aliakmon; 16. Paliouris, Thyamis; 17. Milea, Aoos; 18. Olosson, Mavrorema; 19.
Kalipeuki, Skamnias; 20. Smokovo, Onohonos; 21. Kaitsa, Onohonos; 22. Gorgopotamos Bridge, Gorgopotamos; 23. Gorgopotamos
Village, Gorgopotamos; 24. Dimosaris, Dimosaris; 25. Adias, Adias; 26. Platanistos, Platanistos; 27. Reumata, Aspropotamos; 28.
Piros, Prevedos; 29. Karytena, Alfeios; 30. Tsouraki, Tsouraki; 31. SL98, Tsouraki; 32. Methydrio, Stenon; 33. Gortys, Lousios;
34.Marina, Neda; 35.Elea, Neda; 36.Kalonero, Peristeria; 37. Artiki, Peristeria; 38.Vrachopanagitsa, Pamisos; 39.Ag.Floros, Pamisos;
40.Aris, Pamisos; 41.Messini, Pamisos; 42. Apolakkia, Sianitis; 43. Gadouras, Gadouras; 44. Egkares, Egkares; 45. Amphilissos,
Amlissos; 46. Pyrgos, Amphilissos; 47. Manolates, Kokorrema; 48. Ampeliko, Vourkou.
203
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205
and 0.71, respectively). Therefore, slope, total
hardness, alkalinity, and nitrite were removed
prior to CCA. Hence, from the 40 environmental
variables, 35 were retained. Eleven of the 35 envi-
ronmental variables were signicant (p 0.05) in
explaining species variation as derived from CCA.
Total variance in species abundance data was 4.214
and the sum of all canonical eigenvalues 3.519
(Table 2). The percentage of the total variation of
bug species explained by the environmental vari-
ables accounted thus for 83.5% (3.519 100/
4.214).
The relationship of Heteroptera species with the
11 environmental variables is presented in Figure 3
(see Electronic Supplementary Material
1
). The rst
ordination axis (horizontal axis) reected a gradi-
ent mostly related to forests, aquatic macrophytes
(emergent and submerged), cropland and chloride.
Forest abundance decreased from the left towards
to right end of the axis. The second axis (vertical
axis) indicated that phosphate and stream width
had the next largest eect on Heteroptera occur-
rence. Chloride ions, aquatic macrophytes and
cropland decreased from the right toward the left
end of the axis.
On the upper right of the ordination diagram,
Sigara dorsalis, Nepa cinerea, Sigara striata,
Hesperocorixa sahlbergi, Micronecta poweri,
Micronecta scholtzi, Sigara falleni, Plea minutiss-
ima and Ranatra linearis were associated with
phosphate, open grassland/bushlands, stream
width, chloride and cropland. The rst four species
exhibited very strong associations with phosphate
and stream width, while they were less inuenced
by the rest of the variables. The remaining species
showed closer associations with cropland and
chloride ions. On the bottom right quadrant,
Notonecta glauca and Ilyocoris cimicoides were
associated with aquatic macrophytes and reeds.
Table 2. Results of the CCA analyses between environmental variables and Heteroptera species. Total inertia is the total variance in
species abundance data
CCA axes 1 2 3 4 Total inertia
Eigenvalues 0.578 0.465 0.425 0.383 4.214
Speciesenvironment correlations 0.968 0.97 0.959 0.945
Cumulative percentage variance
Of species data 13.7 24.7 34.8 43.9
Of speciesenvironment relation 16.4 29.6 41.7 52.6
Sum of all eigenvalues 4.214
Sum of all canonical eigenvalues 3.519
1
Electronic supplementary material is available for
this article at http://dx.doi.org/10.1007/s10750-006-
0274-1 and accessible for authorised users
Figure 2. Hierarchical cluster analysis presenting groups of bug species with similar assemblages.
206
Forests (deciduous, coniferous and mixed
native), altitude and water temperature presented
close relationships with Sigara nigrolineata,
Microvelia reticulata, Microvelia pygmaea, Velia
pelagonensis, Velia caprai, Gerris lacustris, Aquarius
najas, Velia currens, Hydrometra stagnorum and
Limnoporous rufoscutellatus. Forests however,
displayed the highest association with the bug
species of the respective quadrant. All forested sites,
except three, were accompanied with signicant
proportion of boulders and gravel and with low
proportion of aquatic vegetation. Finally, the upper
left quadrant reected the relationship of latitude
with Notonecta maculata and Aquarius paludum.
Variance partitioning
Twelve runs of partial CCA were performed to
explain variation in bug species data by the three
groups of environmental data and their combina-
tions (Table 3). The pure (independent) eect of
local (40.1%), regional (16.8%) and geographic
(7.2%) variables was acquired. Moreover, the pure
eect of joint regional and geographic (25.1%),
geographic and local (48.6%) and local and re-
gional (71.4%) variables were obtained. Using the
results of Table 3 and the hypothetical model of
Qinghong (1997), total explained variation was
partitioned into seven parts (Table 4). Thus, the
nal equation, in terms of percentage of variation
(total inertia 100/4.214), can be written as shown
in Table 5.
Common variation (the joint fraction of the
three environmental groups) accounted for only
3% (total inertia 100/3.519, same equation
applies for the following components) of the total
explained variation. The joint eect of local and
regional variables accounted for 17%, whereas
for local and geographic 1.5%. The joint eect of
regional and geographic accounted also approxi-
mately 1.5%. Pure local variables were the
main source of variation accounting for 48%
of the total explained variation followed by
regional (20%) and geographic (8.5%) variables.
Unexplained variation accounted for 16.5%
(10083.5) (see Fig. 4).
Figure 3. CCAplot showing the relationship of the 23 bug species (+) withthe signicant environmental variables. First axis is horizontal
and second axis vertical. Codes for both species and environmental variables are shown in Electronic Supplementary Material.
207
Discussion
It is well documented that some bug species
display distinct habitat preferences (Macan, 1954;
Savage 1982, 1994a, b) while others are capable to
inhabit a wide range of habitats, from rock pools
to more complex habitats such as lakes (Savage,
1989; Kurzatkowska, 1993; Svensson et al., 2000).
Some bugs can tolerate environmental conditions
that would be lethal to other invertebrate species,
Table 3. Variation partitioning by partial canonical correspondence analysis (CCA) of Heteroptera distribution explained by three
groups of environmental variables, geographical (Geo), local (Loc) and regional (Reg). The total inertia used here is the sum of all
canonical eigenvalues. The sum of all eigenvalues in a correspondence analysis of the species matrix is 4.214. Thus, the total percentage
of the total variation of bug species matrix for each step is: total inertia 100/4.214
Run Responder Environmental variables Covariable Total inertia % Variation
Total eect: all environmental variables
Species All groups 3.519 83.5
Partial eect 1 Combination: Loc and Reg & Geo
1 Species Loc Reg & Geo 1.691 3.519 40.1
2 Species Reg & Geo 1.828 43.4
3 Species Reg & Geo Loc 1.059 3.519 25.1
4 Species Loc 2.460 58.4
Joint eect: Loc M Reg & Geo = 2.460 )1.691 = 1.828 )1.059 = 0.769 18.2
Partial eect 2 Combination: Reg and Geo & Loc
1 Species Reg Geo & Loc 0.708 3.519 16.8
2 Species Geo & Loc 2.811 66.7
3 Species Geo & Loc Reg 2.047 3.519 48.6
4 Species Reg 1.472 34.9
Joint eect: Reg M Geo & Loc = 2.811 )1.472 = 2.047 )0.708 = 1.339 31.8
Partial eect 3 Combination: Geo and Loc & Reg
1 Species Geo Loc & Reg 0.302 3.519 7.2
2 Species Loc & Reg 3.217 76.3
3 Species Loc & Reg Geo 3.011 3.52 71.4
4 Species Geo 0.509 12.1
Joint eect: Geo M Loc & Reg = 3.217 )0.509 = 3.011 )0.302 = 2.708 64.3
Table 4. Total explained variance of Heteroptera distribution calculated by using the results of Table 3 and the hypothetical model of
Qinghong (1997)
Part Environmental variables & covariables Equations Results
(1) Joint eect of Loc and [Geo and Reg] A + B + C = 0.769
(2) Joint eect of Reg and [Loc and Geo] A + C + D = 1.339
(3) Joint eect of Geo and [Loc and Reg] B + C + D = 2.708
(4) Pure Loc + pure Reg + joint eect
of Loc and Reg
1.691 + 0.708 + A = 3.011 Joint eect of Loc and Reg = A = 0.612
(5) Pure Loc + pure Geo + joint eect
of Loc and Geo
1.691 + 0.302 + B = 2.047 Joint eect of Loc and Geo = B = 0.054
(6) Pure Reg + pure Geo + joint eect
of Reg and Geo
0.708 + 0.302 + C = 1.059 Joint eect of Reg and Geo = C = 0.049
(7) Joint eect of Geo, Loc and Reg (D)
Total explained variance
(TEV) = D + 0.612 + 0.054 + 0.049 + 0.708 + 0.302 + 1.691 = 3.519
D = 0.103
208
while others show less tolerance. Several Corixid
species were found in acidic mining lakes of
Lusatia, Germany with a pH<3 (Wollmann,
2000) whereas the quality of the environment
(i.e., water pollution and hydromorphological
degradation) inuences the successful colonisation
and populations of the water strider A. najas
(Ahlroth et al, 2003). Heteroptera are most diverse
in warm, heavily vegetated, lentic or slow lotic
waters. A study carried out in Lake Balaton,
Hungary and its adjacent streams, showed greater
species richness and abundance in streams with
environmental heterogeneity (Biro, 2003). In fact,
greatest bug species richness in Greek running
waters was found in lentic streams with rich aquatic
vegetation and with increased nutrient levels.
Streams with restricted microhabitat diversity
(i.e., abiotic substrates such as boulders, gravel,
etc.) supported the least number of dierent spe-
cies, usually those of the Gerridae and Veliidae
families.
Gerridae and Veliidae species were widely
distributed with A. najas and V. caprai inhabiting
most streams of this study. A. najas, as shown
from cluster analysis, occurred frequently with
V. caprai, H. stagnorum and G. lacustris in for-
ested sites at medium altitudes (Fig. 3, Table 1).
Regarding Corixids, S. dorsalis, S.striata and
S. falleni seemed to be closely associated with
each other and were found in sites with simi-
lar environmental characteristics. Similarly,
M. poweri and M. scholtzi also appeared to
inhabit sites with the same characteristics (e.g.,
microhabitat composition). Common eco-groups
Figure 4. Variance partitioning of Heteroptera species data. (A) Represents the bulk variation in original Heteroptera data explained
by the three environmental groups (Bu) and the unexplained (Uv) variation; (B) Represents the pure eect of local (a), regional (b),
geographic (c) and unexplained variables (d) (d = {a + b + c} )Bu); (C) Represents the joint eects of regional and geographic (bc),
geographic and local (ca) and local and regional (ab) variables; (D) Represents the joint eects of the total explained variance using
data from Table 3 and the hypothetical model of Qinghong (1997), (ab) Local and regional, (ac) local and geographic and (cb) regional
and geographic variables and nally (E) Represents the pure eects of the total explained variance using of local (a), regional (b) and
geographic (c) variables.
Table 5. Table presenting the nal equation of the total explained variance (TEV) of Heteroptera distribution. TEV = Joint
variation + partial joint variation + unique variation
Joint variation Partial joint variation Unique variation TEV
Reg, Geo, Loc Loc and Reg Reg, Geo, Loc
Loc and Geo
Reg and Geo
2.4 14.5 + 1.2 + 1.2 16.8 + 7.16 + 40.1 83.50%
Loc, Local; Geo, Geographic; Reg, Regional.
209
have been established in Hungary by Hufnagel
et al. (1999), which are not very distinct from
those of this study.
The results of the ordination analysis
revealed the signicant environmental variables
that were the major explanatory variables struc-
turing Heteroptera assemblages. Land use/cover,
microhabitat composition, stream size, water
chemistry and geographic position were the most
important variables along the four CCA axes in
explaining Heteroptera variation. The signicant
association of water chemistry with bug species is
well documented (Savage, 1982, 1989, 1994a;
Jansson, 1987). Some Corixids have been shown to
be associated with organic pollution (Biesiadka &
Tabaka, 1990) but also with natural eutrophic
conditions (Savage, 1982, 1994a). Water body-size
has been perceived as a signicant factor deter-
mining Heteroptera assemblages and distribution,
especially for Corixidae (Macan, 1954; Savage,
1994a). Relationships between water body-size
and bug species have been detected by Hufnagel
et al. (1999) who distinguished Heteroptera pref-
erences for habitat types ranging from small
shallowwaters tolarge deepwaters. Bug preferences
are not constrained to habitat types. Microhabitat
composition (e.g., aquatic vegetation, sand, gravel,
etc.) is of major importance to Heteroptera assem-
blages anddistribution(Macan, 1938, 1939; Savage,
1989; Garcia-Aviles, 1996). Distribution patterns of
this insect group are well dened in some countries
(Savage, 1990; Popham, 1949; Macan, 1939), how-
ever limited information is available regarding
Heteroptera associations directly with larger scale
variables such as land use/cover.
The upper and bottom right of the ordination
reected the distribution of species in relatively
large and deep running or standing waters with
rich aquatic vegetation and nutrients. Most
streams on the right of the ordination ow
through agricultural basins, which consequently
lead in most cases to the increase of nutrient and
organic (municipal and farm waste) concentra-
tions. Corixidae and Pleidae species were mainly
found in pool waters with the aforementioned
characteristics. The bottom left of the ordination
diagram reected species found in smaller sized
(width and depth) streams, with abundant forests
at relatively higher altitudes and with colder
waters. Forests however, were the most important
variable and seemed to be mostly associated with
the species of the respective quadrant. Veliidae and
Gerridae species of this quadrant were collected
either at the pool zones of the sampling reach or at
faster currents. A. najas, G. lacustris and V. caprai
were generalists since they were found almost in all
stream habitat types, however their abundances
were greater in forested streams. Finally,
geographic position (latitude) was associated with
species of the upper left quadrant, especially
N. maculata.
Partial CCA indicated that local variables play
a major role in Heteroptera variation while
geographical position appears to be the least
inuencing factor. Land use/cover was the second
most important environmental factor determining
species variation. It is well known that regional
and local variables are interrelated since stream
hydromorphology and quality are inuenced by
land use/cover (Vannote et al., 1980; Allan, 2004;
Sandin & Johnson, 2004). In fact, the joint eect of
local and regional variables accounted for 17% of
the total explained variation. In contrast, when
geographical variables were combined either with
local or regional variables, variation loading
accounted for 1.5%, respectively. This suggests
that geographic location is perhaps less important
to Heteroptera species variation, possibly due to
the migration and dispersal abilities of this insect
group (Popham, 1964).
Concluding, it should be taken into account
that some sites were sampled during three seasons
while others during two or only one. This could
have inuenced to some extent the results of this
study, as some species were probably missed if
they occurred in another season than the one
sampled. In addition, those sites sampled more
than one season and with an extra sampling
method (RIVPACS) were likely to comprise more
dierent species (individuals) than those sites
sampled once and with only one method simply
due to the higher number of individuals being
sampled. Nevertheless, this study provided a rst
clue of the inuence of spatial and environmental
components on Heteroptera species and the key
abiotic (environmental) variables structuring
Heteroptera assemblages in Greek running waters.
Moreover, distribution patterns of Heteroptera
species were described along with several selected
ecological preferences (Table 1).
210
Conclusions
At regional scale, assemblages were mainlydividedin
forested and agricultural landscapes, which conse-
quently inuence the quality and hydromorpholog-
ical condition of the receiving waters (Allan et al.,
1997; Allan, 2004). At local scale, species distribution
was separated according to water quality and
microhabitat composition, except commonbugs that
were widely distributed. Local, followed by regional
variables, were the main environmental factors
determining Heteroptera assemblages while geo-
graphic position exhibited the least inuence. These
ndings denote that some bug species could possibly
be used for biomonitoring purposes, as changes to
local and/or regional parameters are likely to aect
their assemblage structure. Understandingbothlocal
and regional-scale parameters is essential for
explaining the factors that structure aquatic and
semi-aquatic bug assemblages.
Acknowledgements
The data of this work were collected within the
framework of the AQEM [EVK 1-CT1999-00027]
and STAR [EVK-CT-2001-00089] projects funded
by the European Commission, 5th Framework
Program, Energy, Environment and Sustainable
Development, Key Action 1: Sustainable Manage-
ment and Water Quality and by the General
Secretariat for Research and Technology, Ministry
of Development, Greece. We would also like to
thank Dr Anthony Polwart and Mrs Hazel Hulme
from Keele University for providing us some
important articles as well as Dr Steven Declerck
and an anonymous referee for their valuable
comments and suggestions on this manuscript.
References
Ahlroth, P., R. V. Alatalo, A. Holopainen, T. Kumpulainen &
J. Suhonen, 2003. Founder population size and number of
source populations enhance colonization success in water
striders. Oecologia 137: 617620.
Allan, J. D., D. L. Erickson & J. Fay, 1997. The inuence of
catchment land use on stream integrity across multiple spa-
tial scales. Freshwater Biology 37: 149161.
Allan, J. D., 2004. Landscapes and riverscapes: the inuence of
land use on stream ecosystems. Annual Reviews of Ecology
and Systematics 35: 257284.
AQEM Consortium, 2002. Manual for the application of
the AQEM method. A comprehensive method to assess
European streams using macroinvertebrates, developed for
the purpose of Water Framework Directive. Version 1.0,
February 2002.
Armitage, P. D., D. Moss, J. F. Wright & M. T. Furse, 1983.
The performance of a new biological water quality score
system based on macroinvertebrates over a wide range of
unpolluted running water sites. Water Research 17: 333347.
Biesiadka, E. & K. Tabaka, 1990. Investigations on water bugs
(Heteroptera) of Szczytno Lakes. Fragmenta Faunistica 33:
4569.
Biro, J., 2003. Temporal-spatial pattern of true bug assemblages
(Heteroptera: Gerromorpha, Nepomorpha) in Lake Balaton.
Applied Ecology and Environmental Research 1: 173181.
Borcard, D., P. Legendre & P. Drapeau, 1992. Partialling out
the spatial component of ecological variation. Ecology 73:
10451055.
Brown, E. S., 1948. A contribution towards an ecological
survey of the aquatic and semi-aquatic HemipteraHet-
eroptera (water bugs) of the British Isles; dealing chiey
with the Scottish Highlands and East and South England.
Transactions of the Society for British Entomology 9:
151195.
Drosopoulos, S., 1980. Hemipterological studies in Greece. Part
1 Heteroptera. A catalogue of the reported species. Bio-
logia Gallo-Hellenica 9: 163185.
Eyre, M. D. & G. N. Foster, 1989. A comparison of aquatic
Heteroptera and Coleoptera communities as a basis for
environmental and conservation assessments in static water
sites. Journal of Applied Entomology 108: 355362.
Garcia-Aviles, J., M. A. Puig & A. G. Soler, 1996. Distribution
and association of the aquatic Heteroptera of the Balearic
Islands (Spain). Hydrobiologia 324: 209217.
Hill, M. O., 1979. DECORANA A FORTRAN Program for
Detrended Correspondence Analysis and Reciprocol Aver-
aging. Ecology and Systematics, Cornell University, New
York.
Hufnagel, L., G. Bakonyi & T. Va sa rhelyi, 1999. New approach
for habitat characterization based on species lists of aquatic
and semi-aquatic bugs. Environmental Monitoring and
Assessment 58: 305316.
Hutchinson, G. E., 1993. A Treatise on Limnology: Vol. 4. The
Zoobenthos. John Wiley & Sons, Inc., New York .
Illies, J., 1978. Limnofauna Europaea. A Checklist of the
Animals Inhabiting European Inland Waters, with Accounts
of their Distribution and Ecology. Gustav Fisher Verlag,
Stuttgart.
Jansson, A., 1977. Distribution of Micronectae (Heteroptera,
Corixidae) in Lake Pa ija nne, central Finland: correlation
with eutrophication and pollution. Annales Zoologici Fen-
nici 14: 118124.
Jansson, A., 1987. Micronectinae (Heteroptera, Corixidae) as
indicators of water quality in Lakes Vesijarvi, southern
Finland, during the period 19761986. Biological Research
Reports of the University of Jyva skyla 10: 119128.
Josifov, M., 1959. HemipteraHeteroptera from the Island of
Thasos. Bulgarian Academy of Science. Biology 8: 265
269in Bulgarian.
211
Kurzatkowska, A., 1993. Investigations on the developmental
biology of Notonecta reuteri Hung. and Notonecta glauca L.
(Heteroptera). Acta Hydrobiologia 35: 4148.
Macan, T. T., 1938. Evolution of aquatic habitats with special
reference to the distribution of Corixidae. Journal of Animal
Ecology 7: 119.
Macan, T. T., 1939. A key to the British species of Corixidae
(Hemiptera Heteroptera) with notes on their distribution.
Scientic Publications of the Freshwater Biological Associ-
ation No 1.
Macan, T. T., 1954. A contribution to the study of the ecology
of Corixidae (Hemiptera). Journal of Animal Ecology 23:
115141.
Magnien, P., 2000. Observations and interesting capture of
Heteroptera. New species for France and Greece (Heterop-
tera: Corixidae, Nabidae, Tingidae, Lygaeidae, Miridae,
Cydnidae). Nouvelle Revue DEntomologie 6: 284287.
McCaerty, W., 1981. Aquatic Entomology. Science Books
International, Inc, Boston.
Moreno, J. L., A. Milla n, M. L. Sua rez, M. R. Vidal-Abarca &
J. Velasco, 1997. Aquatic Coleoptera and Heteroptera
assemblages in waterbodies from ephemeral coastal streams
(ramblas) of south-eastern Spain. Archives fur Hydrobi-
ologie 141: 93107.
Papacek, M., 2001. Small aquatic and ripicolous bugs (Het-
eroptera: Nepomorpha) as predators and prey: the question
of economic importance. European Journal of Entomology
98: 112.
Petrakis, P. V. & V. Roussis, 2001. Bioindication value of
Hellenic aquatic Heteroptera: an algorithmic approach.
Proceedings of the 10th Pan-Hellenic Congress of Ichthyol-
ogists, Chania, Greece, pp 301304.
Popham, E. J., 1949. A contribution towards an ecological
survey of the aquatic and semi-aquatic HemipteraHet-
eroptera (water bugs) of the British Isles. The Ribble Valley
(Lancashire South and Mid). Transactions of the Society for
British Entomology 10: 144.
Popham, E. J., 1950. Water bugs (HemipteraHeteroptera) of
North Surrey. Journal of the Society for British Entomology
3: 158173.
Popham, E. J., 1964. The migration of aquatic bugs with special
reference to Corixidae (Hemiptera, Heteroptera). Archives
fur Hydrobiologie 50: 450496.
Qinghong, L., 1997. Variation partitioning by partial redun-
dancy analysis (RDA). Environmetrics 8: 7585.
Sandin, L. & R. K. Johnson, 2004. Local, landscape and re-
gional factors structuring benthic macroinvertebrate assem-
blages in Swedish streams. Landscape Ecology 19: 501514.
Savage, A. A., 1982. Use of water boatmen (Corixidae) in the
classication of lakes. Biological Conservation 23: 5570.
Savage, A. A., 1989. Adults of the British aquatic Hemiptera:
key with ecological notes. FBA Scientic Publication 50.
Savage, A. A., 1990. The distribution of Corixidae in lakes and
the ecological status of the North West Midland Meres.
Field Studies 7: 516530.
Savage, A. A., 1994a. The distribution of Corixidae in relation
to the water quality of British lakes: a monitoring model.
Freshwater Forum 4: 3261.
Savage, A. A., 1994b. Corixidae and water quality. Freshwater
Forum 4: 214216.
Skoulikidis, N., Y. Amaxidis, I. Bertahas, S. Laschou & K.
Gritzalis, 2006. Analysis of factors driving stream water
composition and synthesis of management tools a case
study on small/medium Greek catchments. The Science of
the Total Environment 362: 205241.
Sla decek, V. & A. Sla deckova , 1994. Corixidae as indicators of
organic pollution. Freshwater Forum 4: 211213.
Svensson, B. G., B. Tallmark & E. Petersson, 2000. Habitat
heterogeneity, coexistence and habitat utilization in ve
backswimmer species (Notonecta spp.; Hemiptera, Noto-
nectidae). Aquatic Insects 22: 8198.
Tamanini, L., 1979. Guide per il riconoscimento delle specie
animali delle acque interne I-taliane. Eterotteri acquatici
(Heteroptera: Gerromorpha, Nepomorpha). No 6. Consiglio
Nazionale delle Ricerche.
Ter Braak, C. J. F., 1986. Canonical correspondence analysis: a
new eigenvector technique for multivariate direct gradient
analysis. Ecology 67: 11671179.
Ter Braak, C. J. F. & C. Prentice, 1988. A theory of gradient
analysis. Advances in Ecological Research 18: 271317.
Ter Braak, C. J. F. & P. Smilauer, 2002. CANOCO, Software
for Canonical Community Ordination (ver. 4.05). Centre for
Biometry, Wageningen, The Netherlands.
Vannote, R. L., G. W. Minshall, K. W. Cummins, J. R. Sedell
& C. E. Cushing, 1980. The river continuum concept.
Canadian Journal of Fisheries and Aquatic Sciences 37: 130
137.
Wollmann, K., 2000. Corixidae (Hemiptera, Heteroptera) in
acidic mining lakes with pH 3 in Lusatia, Germany. Hyd-
robiologia 433: 181183.
212