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Chief editor: F. Malard
UMR CNRS 5023 - Ecologie des Hydrosystèmes Fluviaux
Université Claude Bernard - Lyon 1 - Bat. Forel (403)
43, boulevard du 11 Novembre 1918
69622 Villeurbanne Cedex, France
e-mail: malard@univ-lyon1.fr
Associate editors: M.-J. Dole-Olivier, J. Mathieu, and F.
Stoch
The following scientists contributed to the production of
the manual: C. Boutin, A. Brancelj, A.I. Camacho, F. Fiers,
D. Galassi, J. Gibert, T. Lefebure, P. Martin, B. Sket, and
A.G. Valdecasas.
Forewords
This sampling manual is published within the framework of
the European project PASCALIS. The project PASCALIS addresses
a problem of growing concern in Europe, i.e. the biodiversity
and ecosystem aspects of groundwater conservation. The main
goal of this project is to establish a rigorous and detailed
protocol for assessing groundwater biodiversity and to develop
operational tools for its conservation. Knowledge on the distribution
of groundwater animals in southern Europe is presently being
organized in a large database and represented on maps. In
addition, groundwater biodiversity is being surveyed at 6
selected regions in Europe following a standardized sampling
procedure. Most taxonomic groups inhabiting a wide range of
groundwater habitats are considered. The present manual provides
a detailed description of the standard protocol that is used
to assess groundwater biodiversity in the selected regions.
This manual is made available below so
that the project PASCALIS benefits for multiple end-users
including scientists, public and civil society, groundwater
managers, and policy makers. Comments regarding the content
of this manual can be addressed to the public
forum of the web site PASCALIS.
The project leader
Prof. Janine Gibert
Content
of the manual (download
here)
Introduction
Part I: Biodiversity in ground
water
 1.1. Diversity
of groundwater habitats
1.2. Diversity
of groundwater fauna
Part II: Objectives of the sampling
manual
2.1. Objectives
2.2. Limitations
Part III: Hierarchical sampling
scheme
3.1. Sampling
strategy
3.2. Sampling
procedure
3.3. Definition
of sampling units and habitats
3.3.1. Region
3.3.2. Hydrogeographic
basins
3.3.3. Karstic
ground water
3.3.3.1.
The unsaturated (vadose) zone
3.3.3.2.
The saturated (phreatic) zone
3.3.4. Subsurface
water in unconsolidated sediments
3.3.4.1.
The hyporheic zone
3.3.4.2.
Ground water in unconsolidated sediments
Part IV: Selection of sampling
sites
4.1. Site
selection
4.1.1. Sampling
sites as access points to the different units of a region
4.1.2. Pre-selection
of sampling sites
4.1.3. Field
surveys
4.2. Register
of sampling sites
4.3. Mapping
of sampling sites
4.4. Environmental
features of habitats
4.4.1. Site
description
4.4.2. Land
use and human activities
4.4.3. Physico-chemical
measurements during sampling
Part V: Sampling methods and
devices
5.1. Sampling
in the hyporheic zone
5.1.1. Data
sheet
5.1.2. Measurement
of vertical hydraulic gradient
5.1.3. The
Bou-Rouch pumping method
5.2. Sampling
in springs
5.2.1. Data
sheet
5.2.2. Drifting
fauna
5.2.3. Sampling
of habitats near springs
5.2.4. Artificial
substrates and baited traps
5.3. Sampling
in caves
5.3.1. Safety
5.3.2. Data
sheet
5.3.3. Methods
for sampling slowly infiltrating water in the vadose zone
5.3.3.1.
Filtering water dripping from cave ceiling
5.3.3.2.
Sampling in gours, puddles and pools
5.3.4. Methods
for sampling subterranean brooks, rivers, lakes and siphons
5.4. Sampling
in wells
5.4.1. Well
type and design
5.4.2. Data
sheet
5.4.3. Phreatobiological
net
5.4.4. Baited
traps
5.4.5. Pumping
of well water
5.4.5.1.
Sampling method
5.4.5.2.
Permanent pump
5.4.5.3.
Suction pumps for the sampling of shallow water-table aquifers
5.4.5.4.
Pressure pumps for the sampling of deep water-table aquifers
5.5. Methods
for physico-chemical measurements
5.5.1. Collection
and preservation of samples
5.5.2. Field
measurements
5.5.3. Laboratory
analyses
5.6. Fixation,
preservation, and processing of faunal samples
5.6.1. Fixation
and preservation of most taxonomic groups
5.6.2. Specific
fixation and preservation procedures
5.6.3. Processing
of faunal samples
Conclusion
Acknowledgements
References
List of figures
List of tables
List of photos
Appendixes (not available on this site)
Introduction
Although research on biological diversity in subsurface water
typically lags behind that of surface freshwater, considerable
efforts by scientists over the second half of the 20th century
have revealed the unexpectedly high diversity of living forms
in ground water (Botosaneanu 1986, Juberthie and Decu 1994,
1998). During the 1990s, there have been more than 10 books
dealing with the biology and ecology of ground water, a publication
rate probably unequalled in many other research fields (Botosaneanu
1986, Camacho 1992a, Juberthie and Decu 1994, 1998, Gibert
et al. 1994a, 1997, Culver et al. 1995, Wilkens et al. 2000,
Jones and Mulholland 2000, Griebler et al. 2001). Whereas
earlier research works were essentially restricted to the
study of animals living in caves, faunal investigations carried
out in a variety of subterranean habitats have demonstrated
that groundwater animals can potentially be found wherever
subsurface water exists (Rouch 1986, Danielopol 1989). Despite
a long tradition of species inventory in subterranean ecology,
groundwater ecologists have only recently begun to synthesise
and map the vast amount of existing data on the distribution
of groundwater species (Juberthie and Decu 1994, 1998). Mapping
biodiversity is emerging as an essential task for understanding
biodiversity patterns and the development of conservation
strategies. Culver et al. (2000) mapped the distribution of
927 obligate cave-dwelling species in the 48 contiguous states
of the United States (www.karstwaters.org).
In Europe, the research project PASCALIS “Protocols for
the ASsessment and Conservation of Aquatic Life In the Subsurface”
is the first attempt to provide large-scale distribution patterns
of specific richness and endemism in ground water. The first
phase of the project is devoted to the compilation and mapping
of existing data on the distribution of subterranean aquatic
taxa across a European gradient (Belgium, France, Spain, Italy,
and Slovenia).
In addition to the mapping of
groundwater biodiversity, the project PASCALIS also aims to
develop several validated methods for:
1) determining the reliability of biodiversity patterns revealed
by the mapping of existing data;
2) using a standard field sampling protocol to obtain an unbiased
estimate of groundwater biodiversity in areas for which no
data exist;
3) predicting overall species richness based on biodiversity
indicators in regions with incomplete data sets.
This second phase of the project is essential because the
assessment of biodiversity within a given area strongly depends
on:
1) the number of observations (i.e. sampling sites);
2) the number of habitats (e.g. historically, sampling was
mainly carried out in caves);
3) the amount of information available on various taxonomic
groups (e.g. the macrofauna has received considerably more
attention than the meiofauna).
These three important issues have been largely overlooked
in the field of groundwater biodiversity, thereby severely
restricting the usefulness of biodiversity maps for basic
and conservation purposes. Within the framework of the project
PASCALIS, these issues will be addressed based on the results
of an extensive sampling survey in 6 regions distributed in
southern Europe: the Walloon karst (Belgium), the meridional
Jura (Eastern France), the Roussillon region (France), the
Cantabrica (Spain), the Lessinian mountains (Italy), and the
Krim massif (Slovenia). Faunal sampling within these regions
will be carried out following a standard protocol, the objective
of which is to provide a data set as complete as possible
on the distribution of groundwater species.
The ensuing material provides
a detailed description of the sampling protocol that will
be applied in the selected regions. This protocol has been
established within the framework of the research project PASCALIS
based on the results of:
1) discussions among partners of the project;
2) an electronic forum (11 February - 4 March 2002) among
37 scientists from 11 countries;
3) a sampling workshop
held in Lyon, France (22-26 April 2002) during which 41 participants
from 6 European countries learnt about methods and devices
for sampling groundwater fauna.
The first part of the manual gives a brief account of the
diversity of habitats and animals in subsurface water. Secondly,
we define the objectives of the sampling protocol and specify
the geomorphologic features of areas where this protocol can
be applied. Thirdly, we present the rationale of a stratified
sampling scheme and define the different hierarchical units
to be sampled. Fourthly, we propose a general procedure for
selecting and mapping sampling sites and managing environmental
data. The last part of the manual outlines the recommended
methods and devices for the sampling of groundwater fauna
in springs, caves, wells, and the hyporheic zone of rivers.
List
of figures
Figure 1: Three-dimensional view of different aquifers. Modified
after Gibert (2001).
Figure 2: A classification of groundwater fauna
(modified after Gibert et al. 1994b).
Figure 3: The interstitial habitat and some
of the subterranean-dwelling organisms. After Danielopol et
al. (1994).
Figure 4: Hierarchical sampling scheme used
within the framework of the European project PASCALIS (Protocols
for the ASsessment and Conservation of Aquatic Life In the
Subsurface).
Figure 5: Different steps of the sampling procedure
used within the framework of the European project PASCALIS
(Protocols for the ASsessment and Conservation of Aquatic
Life In the Subsurface).
Figure 6: Location of the 6 regions selected
for sampling groundwater fauna within the framework of the
European project PASCALIS (Protocols for the ASsessment and
Conservation of Aquatic Life In the Subsurface).
Figure 7: Location of the 4 hydrogeographic
basins selected for sampling groundwater fauna in the Walloon
Karst (Belgium).
Figure 8: Location of the 4 hydrogeographic
basins selected for sampling groundwater fauna in the Meridional
Jura (France).
Figure 9: Location of the 4 hydrogeographic
basins selected for sampling groundwater fauna in the Roussillon
Region (France).
Figures 10.1 and 10.2: Location of the 4 hydrogeographic
basins selected for sampling groundwater fauna in the Cantabrica
Region (Spain).
Figure 11 : Location of the 4 hydrogeographic
basins selected for sampling groundwater fauna in the Lessinian
Mountains (Italy).
Figure 12: Location of the 4 hydrogeographic
basins selected for sampling groundwater fauna in the Krim
Massif (Slovenia).
Figure 13: Idealised view of a karst aquifer.
Modified after Mangin (1974/1975).
Figure 14: The epikarst. A – after Mangin
(1974/1975). B – after Williams (1983).
Figure 15: Conceptual groundwater flow models
in karst aquifers. Modified after Gibert et al. (1994 c).
Figure 16: Surface-subsurface hydrological exchanges
in the hyporheic zone induced by spatial variation in streambed
topography and sediment permeability. Modified after Malard
et al. 2002.
Figure 17: Conceptual cross-sectional models
of surface channels and beds showing relationships of channel
water to hyporheic, groundwater, and impermeable zones. Modified
after Malard et al. (2000).
Figure 18: Spatial and temporal domain of hyporheic
interactions and relation to roughness features in channels.
After Harvey and Wagner (2000).
Figure 19: Geomorphologic units generating hyporheic
flow paths.
Figure 20 (not available yet): Map showing the
location of sampling sites in the Walloon karst (Belgium).
Figure 21: Map showing the location of sampling
sites in the Meridional Jura (France).
Figure 22 (not available yet): Map showing the
location of sampling sites in the Roussillon region (France).
Figure 23: Map showing the location of sampling
sites in the Cantabrica Region (Spain).
Figure 24 : Map showing the location of sampling
sites in the Lessinian Mountains (Italy).
Figure 25: Map showing the location of sampling
sites in the Krim Massif (Slovenia).
Figure 26: Description of the T-bar (left box)
and measurement of pressure differential between surface water
and hyporheic water (right box).
Figure 27: Description of the Bou-Rouch pumping
method for sampling invertebrates in the hyporheic zone of
rivers. Modified after Bou (1974).
Figure 28: Description of the peristaltic pump
for pumping water in the Bou-Rouch stand pipe for the measurement
of physico-chemical parameters and collection of water samples.
Figure 29: Description of the vacuum pump for
sampling invertebrates in the hyporheic zone of rivers. A
manual diaphragm pump is used to create vacuum in the jar.
Figure 30: Drift net for the sampling of groundwater
invertebrates in caves and springs. Modified after Vervier
(1988).
Figure 31: Pond net, Hess sampler, and Surber
sampler for the sampling of groundwater invertebrates in the
benthic layer of springs. Modified after Niederreiter (2002)
and Camacho (1992).
Figure 32: Device for filtering water dropping
from cave ceiling. Animals are trapped in the bottle.
Figure 33: The Karaman-Chappuis method for sampling
groundwater invertebrates in the bank sediments of surface
and underground rivers and lakes. Modified after Delamare
Debouteville (1960).
Figure 34: Artificial substrate for the sampling
of invertebrates in water bodies of caves and limnocrene springs.
After Vervier (1990).
Figure 35: A - Design of a drilled well in unconsolidated
sediments. B – Vertical change in dissolved oxygen concentration
in the screened and unscreened sections of the well.
Figure 36: The Cvetkov phreatobiological net
for sampling groundwater invertebrates in large diameter wells.
Modified after Bou (1974).
Figure 37: Examples of baited traps for sampling
groundwater invertebrates in cave water bodies and wells.
Modified after Boutin and Boulanouar (1983).
Figure 38: Inertial pump for sampling water
and invertebrates in small-diameter wells. Modified after
Malard et al. (1997).
Figure 39: Air-lift pump for sampling invertebrates
in wells. Modified after Malard et al. (1997).
Figure 40: Pneumatic piston pump for sampling
water and invertebrates in wells. Modified after Niederreiter
(2002).
Figure 41: Ejector pump for sampling invertebrates
in wells. Modified after Malard et al. (1997).
List
of tables
Table 1: Comparison of aquatic subterranean habitats based
on the size of the voids, degree of interconnectedness between
voids, and strength of hydrologic linkages with the surface
environment.
Table 2: Species richness among different taxonomic
groups in ground water. After Gibert and Deharveng (2002).
Table 3: Species richness among different taxonomic
groups in European ground water. Extracted from Sket (1999),
Table I.
Table 4: Ranges of specific yield (i.e. effective
porosity) for different geological formations. Modified after
Castany (1982).
Table 5: Ranges of permeability for different
geological formations. Modified after Freeze and Cherry (1979).
Table 6: Structure of the register of sampling
sites used within the framework of the European project PASCALIS
(Protocols for the ASsessment and Conservation of Aquatic
Life In the Subsurface).
Table 7: Data sheet for sampling invertebrates
in the hyporheic zone.
Table 8: Data sheet for sampling invertebrates
in springs.
Table 9: Data sheet for sampling invertebrates
in caves.
Table 10: Data sheet for sampling invertebrates
in wells.
Table 11: Land use categories as defined by
the Corine Land Cover nomenclature.
Table 12: Comparison of several pressure pumps
for sampling invertebrates in wells. Modified after Malard
et al. (1997).
Table 13: Example of data sheet for sorting
faunal samples collected in the Meridional Jura (France).
List
of photos
Photo 1: The T-bar for measuring vertical hydraulic gradient
between the surface stream and the hyporheic zone.
Photo 2: Hammering of mobile pipe into the river
bed for sampling the hyporheos.
Photo 3: The Bou-Rouch pumping method for sampling invertebrates in the hyporheic zone of rivers.
Photo 4: Sampling of hyporheic water with the
peristaltic pump for physico-chemical measurements.
Photo 5: The vacuum pump for sampling invertebrates
in the hyporheic zone of rivers.
Photo 6: Drift net for the sampling of invertebrates
in karstic springs.
Photo 7: Filtering of water dropping from cave
ceiling.
Photo 8: A hand pear-shaped pump for emptying
small water bodies in caves. Water is then filtered through
a plankton net.
Photo 9: A hand peristaltic pump for emptying
small water bodies in caves. Water is then filtered through
a plankton net.
Photo 10: Hand nets for collecting invertebrates
from water bodies in caves.
Photo 11: Filtering of cave water bodies with
a hand net.
Photo 12: Baited trap for collecting animals
in water bodies of caves.
Photo 13: Collection of macro-invertebrates
with a suction device in puddles of caves.
Photo 14: Sampling of interstitial invertebrates
with the Karaman-Chappuis method.
Photo 15: Artificial substrate for sampling
groundwater invertebrates in water-bodies of caves.
Photo 16: A manual device for installing piezometers
in areas where the groundwater table is very shallow (i.e.
< 2 m below the soil surface).
Photo 17: A multi-parameter sonde for measuring
vertical changes in temperature, specific conductance, dissolved
oxygen, pH, and redox potential in wells.
Photo 18: The Cvetkov phreatobiological net
for sampling groundwater invertebrates in large diameter wells.
Photo 19: The Bou-Rouch pump for sampling groundwater
invertebrates in shallow water-table aquifers (i.e. groundwater
table not deeper than about 8 m below the ground surface).
Photo 20: The inertial pump for sampling water
and invertebrates in small diameter wells.
Photo 21: The air lift for sampling groundwater
invertebrates in wells.
Photo 22: The pneumatic piston pump for sampling
water and invertebrates in wells.
Photo 23: The ejector pump for sampling groundwater
invertebrates in wells.
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