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ORIGINAL PAPER
The European land leech: biology and DNA-based taxonomy
of a rare species that is threatened by climate warming
U. Kutschera &I. Pfeiffer &E. Ebermann
Received: 12 March 2007 /Revised: 11 June 2007 / Accepted: 21 June 2007 / Published online: 24 July 2007
#Springer-Verlag 2007
Abstract The European land leech Xerobdella lecomtei
was discovered in 1868 and is one of the rarest animals on
Earth. During the 1960s, several individuals of these
approx. 40 mm long, cold-adapted terrestrial annelids that
inhabit the moist soils of birch forests around Graz, Austria,
were investigated. Only one original research paper has
been published on the biology of this species. Between
2001 and 2005, we re-investigated the morphology of
preserved specimens and searched for living individuals in
their natural habitat that appeared to be intact. We found
only one juvenile individual (length approx. 10 mm),
indicating that this local leech population became largely
extinct over the past four decades. The feeding behaviour of
our ‘lonesome George of the annelids’was studied and is
described here in detail. After its death, the Xerobdella
individual was used for chemical extraction and molecular
studies (deoxyribonucleic acid [DNA] barcoding, based on
one gene, the mitochondrial cytochrome coxidase subunit I).
In addition, novel DNA barcodes for a land leech from
Madagascar and a recently discovered species from Europe
were obtained. Our phylogenetic tree shows that X. lecomtei
is not a member of the tropical land leeches (family
Haemadipsidae), as previously thought, but represents a
separate line of descent (family Xerobdellidae). The decline
of the local leech population around Graz correlates with a
rise in average summer temperatures of +3°C between 1961
and 2004. This warming led to a drastic reduction in the
moisture content of the soil where X. lecomtei lives. We
suggest that human-induced climate change without appar-
ent habitat destruction can lead to the extinction of
populations of cold-adapted species that have a low
colonization ability.
Keywords Climate warming .DNA barcoding .
Land leeches .Xerobdella
Introduction
Tropical land leeches (Haemadipsidae) are found in Sri
Lanka, India, Burma, Malaysia, the Islands of the East
Indies, China, Japan, Australia and Madagascar. In these
areas, these aggressive blood-sucking parasites are confined
to habitats that have a heavy rainfall because moist
conditions are necessary for their terrestrial way of life. In
some regions of India, haemadipsids have been described
as a ‘formidable pest’(Bhatia 1975; Sawyer 1986). These
agile annelids suck the blood of mammals (cows, horses
and humans); many tropical places are dominated by land
leeches to the extent that they are almost un-inhabitable.
However, not all terrestrial leaches are tropical; that is, there
are species that live in temperate climates.
In 1868, Georg Ritter von Frauenfeld (1807–1873)
published a short report on the occurrence of a new leech
species that was discovered in Austria (province Steier-
mark) in the moist soil on a mountain, far away from any
freshwater pond or stream. This unusual annelid, the
European land leech (Xerobdella lecomtei), was described
on the basis of three approx. 40 mm long alcohol-preserved
specimens (v. Frauenfeld 1868). Three decades later,
Naturwissenschaften (2007) 94:967–974
DOI 10.1007/s00114-007-0278-3
U. Kutschera (*):I. Pfeiffer
Institute of Biology, University of Kassel,
Heinrich-Plett-Str. 40,
34109 Kassel, Germany
e-mail: kut@uni-kassel.de
E. Ebermann
Institute of Zoology, Karl-Franzens-University of Graz,
Universitätsplatz 2,
8010 Graz, Austria
Penecke 1896 reported that Xerobdella individuals, found
in relatively cold alpine regions, are sometimes attached to
the skin of the montane salamander Salamandra atra.He
concluded that the European land leech is a facultative
blood sucker on terrestrial amphibians. In 1909, Schuster
discovered remnants of partly digested oligochaetes in the
gut of two sectioned Xerobdella individuals and suggested
that the land leech feeds on earthworms.
Between 1922 and ca. 1940, the zoologist Erich Reisinger
(1900–1978) collected several adult land leeches in the
birch forests around Graz (Austria). He analysed the
feeding and reproductive behaviour of this rare annelid
on animals kept in glass jars in the laboratory. Amphibians
(S. atra etc.) were not attacked by hungry Xerobdella, but
the leeches voraciously fed on earthworms and other
terrestrial oligochaetes (Reisinger 1951). In addition, this
author discovered that Xerobdella is a cold-adapted
terrestrial annelid, which prefers temperatures of 10–15°C.
At 25°C, the land leeches rapidly died.
The Xerobdella population around Graz was analysed a
second time between ca. 1960 and 1970. Within this
decade, more than 20 individuals were collected, preserved
in liquid fixatives and used for an anatomical study
(Moosbrugger and Reisinger 1971). After the death of
Reisinger (1978), only four adult Xerobdella individuals
remained in the Institute of Zoology (Karl-Franzens-
University of Graz). In 2001, we wanted to use part of this
stored material for an ongoing deoxyribonucleic acid
(DNA)-barcoding study on European leeches (determina-
tion of mitochondrial cytochrome coxydase subunit I
[COI] gene sequences for identification of species, see
Pfeiffer et al. 2004,2005; Siddall and Budinoff 2005;
DeSalle et al. 2005; Bely and Weisblat 2006; Waugh 2007;
Kutschera 2007). However, we were unable to extract
amplifiable DNA fragments from these poorly preserved
samples. Because only one original report has ever been
published on the biology of Xerobdella (Reisinger 1951),
we engaged in expeditions to the birch forests where the
leeches were collected four decades ago. These extensive
searches for the European land leech were largely unsuc-
cessful. Between 2001 and 2005, we found only one living
juvenile Xerobdella individual. Hence, the Xerobdella
population analysed by Reisinger 1951 became largely
extinct over past decades, although the habitat of this rare
annelid has not been destroyed.
In this report, we describe the morphology of preserved
individuals of the original population and the biology and
feeding behaviour of this single specimen. After its death,
our ‘Lonesome George of the annelids’(Nicholls 2006)
was used for DNA extraction and sequencing to explore the
systematic position of this enigmatic animal as well as its
supposed phylogenetic relationship to the tropical land
leeches (Blanchard 1917; Sawyer 1986; Nesemann and
Neubert 1999). Based on a recent study that documents the
human-induced climate change in Austria over the last four
decades (Kabas 2005), we propose a hypothesis that may
account for the almost complete extinction of this local
population of land leeches.
Materials and methods
Leech specimens and field work
Four formaline-preserved adult specimens of X. lecomtei
from the Reisinger collection were stored without refriger-
ation for time periods from 30 to 40 years. In 2001, these
animals were transferred to 70% ethanol and 2 years later
investigated under a light microscope. One living juvenile
X. lecomtei individual was collected in 2002 in a sample of
relatively moist soil (region around Graz, Austria) and was
maintained in a closed plastic container (approx. 10 × 15 ×
5 cm) at the University of Kassel (Institute of Biology, cold
room; temperature =10–15°C). The terrarium was equipped
with moist filter paper and leaves of birch (Fagus sylvatica)
trees from the leech’s natural habitat (Moosbrugger and
Reisinger 1971). Over a period of 10 months, this
individual was fed nine times by the addition of one
earthworm (adult Lumbricus castaneus or juvenile L.
terrestris, length approx. 30–40 mm). The feeding episodes
were recorded with a camera as previously described
(Kutschera and Wirtz 2001; Kutschera 2003). One repre-
sentative episode was selected and is illustrated by a
schematic drawing. After its death, the Xerobdella individ-
ual was stored in 95% ethanol at −20°C.
Land leeches from Madagascar were collected in the
vicinity of the town of Antananarivo (see map in Goodman
and Benstead 2003). Individual leeches were picked up
from leaves in forests. About 40 individuals, juveniles and
adults, were collected; the length of living leeches at rest
was 5 to 16 mm. All specimens were immersion fixed and
preserved in 95% ethanol. The land leeches were assigned
to the taxon Mallagabdella fallax according to the key of
Richardson (1978). Alcohol-preserved individuals of the
European species Hirudo verbana (Kutschera 2006),
Trocheta pseudodina (Pfeiffer et al. 2005) and Haemopis
elegans (Grosser 2004) were obtained from M. Roth
(Giessen, Germany) and C. Grosser (Leipzig, Germany).
DNA extraction
All leeches were stored in 95% ethanol at −20°C until used
for extraction of total DNA. The caudal sucker was
removed and utilized for all analyses. This organ was
specifically used to minimize the possibility of contamina-
tion from prey (or host) DNA stored in the gastric region of
968 Naturwissenschaften (2007) 94:967–974
the leeches. From these tissues (about 10 to 80 mg per
sample), total DNA was extracted using the QIAamp Tissue
Kit (Quiagen GmbH, Hilden, Germany) as described by
Pfeiffer et al. (2004). The resulting DNA was diluted in
50 μl of double-distilled water and used for further
analyses. All DNA extractions were repeated at least two
times with different specimens (Pfeiffer et al. 2004). In the
case of X. lecomtei, only one juvenile specimen was
available for DNA extractions, which were repeated from
small tissue samples excised from the flanks of the body. It
should be noted that molecular studies based on a single
specimen are not uncommon (for a recent example, see
Giribetetal.2006).
PCR amplification, sequencing and data analysis
We examined sequence diversity in a specific 648-bp
fragment of the mitochondrial COI gene (Pfeiffer et al.
2004). This sequences was amplified using the ‘universal
primers’designed by Folmer et al. (1994): LCO 1490: 5′-
GGTCAACAAATCATAAAGATATTGG-3′and HCO 2198:
5′-TAAACTTCAGGGTGACCAAAAAATCA-3′.
Amplification reactions for COI included 10× polymerase
chain reaction (PCR) buffer (10 mM MgCl
2
, pH 8.3) and Q-
solution, 100 μM of each deoxyribonucleotide triphos-
phate, with 1 U of Taq polymerase and 10 pmol of each
primer and 4 μl of the DNA extract. The total volume of
this solution was 20 μl. The amplifications were carried out
with initial denaturation at 95°C for 10 min, followed by 38
cycles of one denaturation step at 94°C for 40 s, primer
annealing at 38°C for 40 s and primer extension at 72°C for
45 s in a Thermocycler (Biometra, Göttingen, Germany).
Two microliters (20–50 ng) of the PCR products were
cycle sequenced as previously described (Pfeiffer et al.
2004). DNA sequences were analysed on an ABI Prism
3100 genetic analyzer (Applied Biosystems, Foster City,
USA) according to manufacturer’s instructions and were
aligned subsequently by eye in BIOEDIT (Hall 1999).
Sequence divergences among individuals were quantified
using the Kimura-2-parameter distance scheme (Pfeiffer et
al. 2004) and graphically displayed in a neighbour-joining
tree (Saitou and Nei 1987;Waugh2007). The novel
sequences have been deposited in GenBank (accession
nos. EF 125040–EF 125044).
Results
Morphology of adult leeches
Four decades ago, Moosbrugger and Reisinger (1971)
collected, preserved and investigated several adult Xerob-
della individuals. In 2001, only four formaline-preserved
specimens were found in the archives of the Institute of
Zoology (Graz, Austria). Because no photograph of this rare
annelid has ever been published, we document here the
unique morphology of this animal species (Fig. 1). The
formaline-preserved leeches that were transferred for further
conservation into 70% ethanol had a body length of 30–
45 mm and a maximum diameter of 3.0–3.5 mm. In the
individual depicted here (Fig. 1a), the glandular clitellum
covers annuli 26 to 40; the swollen clitellum of a sexually
mature earthworm is shown in Fig. 3. In the head region of
Xerobdella, the oral palps or ‘tenacles’were apparent; these
structures assist in foraging and feeding (Fig. 1b). Four pairs
of dark-pigmented eyes were counted on all individuals
investigated here. In the clitellar region, the male and first
female gonopores are separated by three to four annuli. In
addition, the accessory (second) female gonopore (bursa
porus), a structure of unknown function, is also apparent in
our photograph (Fig. 1c). Nesemann and Neubert (1999)
have pointed out that specific radial furrows within the
posterior sucker of Xerobdella are a characteristic feature of
Fig. 1 Representative adult specimen of the European land leech
(Xerobdella lecomtei), collected in Austria ca. 1960. Ventral view of
entire animal (a); head region with oral palps, ventral view (b);
clitellar region, showing the male and the two female gonopores,
respectively (c), and posterior sucker with characteristic radial furrows
(d). The leech was originally stored in formaline and since 2002
preserved in 70% ethanol. Scale bars=2 mm
Naturwissenschaften (2007) 94:967–974 969
this European annelid that appear to be identical with those
of the tropical land leeches. In our adult specimens, these
structures were detected (Fig. 1d), but the furrows are less
pronounced than in the schematic drawings published by
these earlier investigators.
Behaviour of the land leech
Our collection efforts into the beech forests around Graz
yielded only one juvenile specimen of X. lecomtei, which
was attached to a rotten leaf (Fig. 2a). The length of this
individual was about 10 mm at rest (diameter approx.
1 mm); when fully extended, the leech reached a length of
approximately 18 mm. During the day, the inactive leech
coiled into a ball-like configuration with the head in the
centre of the knot (Fig. 2b). At night (notably after
midnight), the leech was active and, when hungry, regularly
foraged for food (Fig. 2c).
Reisinger (1951) reported that adult hungry Xerobdella
individuals, maintained in captivity, do not feed on
amphibians such as S. atra, but rapidly attack and swallow
small earthworms. However, foraging and feeding behav-
iour of a juvenile land leech has not yet been described.
Nine feeding episodes of our individual (Fig. 2a) were
recorded. A representative chain of events is shown in
Fig. 3. The resting land leech in knotted posture (Fig. 3a) is
alerted by the addition of an earthworm (adult L. castaneus
or juvenile Lumbricus terrestris). The leech detects the
slime trail of the nearby organism by means of the palp-like
tentacles on either side of the mouth (Fig. 1b) and rapidly
attaches to its prey (Fig. 3b). Within a few minutes, the
leech cuts through the epidermis of the earthworm with the
aid of the teeth on its three jaws and sucks body fluids from
the wounds. This feeding episode, during which the
juvenile leech is carried around by the mobile earthworm,
lasts for 2 to 3 h. The thick, satiated leech then leaves its
prey and returns to its knotted posture (Fig. 3c). The
satiated leech remained in this characteristic resting position
until the ingested body fluids were completely digested (ca.
1 week later).
Taxonomic status of Xerobdella
In accordance with earlier investigators, Reisinger (1951)
interpreted Xerobdella, a European sanguivorous annelid,
as a member of the Haemadipsidae (tropical land leeches).
In this context, he regarded Xerobdella as a relict taxon
from the Tertiary (Moosbrugger and Reisinger 1971). On
the basis of nuclear 18 S ribosomal DNA (rDNA) sequence
data, this hypothesis has been questioned (Borda and
Siddall 2004). Because no mitochondrial DNA sequences
were available for Xerobdella, we extracted DNA from our
single specimen (Fig. 2) after its death and obtained a 648-
bp mitochondrial COI gene sequence. In addition, new COI
data were obtained for the following taxa: Malagabdella
fallax, a common tropical land leech from Madagascar
(Richardson 1978), H. verbana, a blood-sucking amphibi-
ous medicinal leech from Turkey (Kutschera 2006,2007)
and two relatively large terrestrial leech species from
Germany, T. pseudodina (Pfeiffer et al. 2005) and the
recently discovered taxon H. elegans (Grosser 2004). These
five newly acquired COI sequences (GenBank accession
nos. EF 125040–EF 125044) were supplemented by
published COI data from the literature, for the following
leech species: Haemadipsa sylvestris and Chtonobdella
bilineata from Vietnam and Australia, respectively
(Richardson 1968; Siddall and Burreson 1998), Haemopis
sanguisuga,Erpobdella octoculata and Trocheta haskonis
from Europe (Pfeiffer et al. 2004,2005).
Our phylogenetic neighbour-joining tree (Fig. 4)yielded
the following information. The three tropical land leeches
(Haemadipsa from Asia, Malagabdella from Madagascar
and Chtonobdella from Australia) form a clade that excludes
the terrestrial Xerobdella from Europe. This rare annelid
represents a separate line of descent, parallel to the haemopid/
hirudinid clade (Haemopis,Hirudo). The amphibious preda-
tory erpobdellid taxa of the genus Trocheta,supplementedby
the aquatic type species E. octoculata, are not closely related
to Xerobdella (outgroup). Hence, X. lecomtei is not a member
of the tropical Haemadipsidae but may be a relative of the
Fig. 2 Juvenile living individual of Xerobdella lecomtei (arrow),
attached to a fallen leaf of birch (Fagus sylvaticus) from its natural
habitat (a). The same leach on moist filter paper in resting position (b)
and alerted by an earthworm (c). Scale bars=10and2mm,
respectively
970 Naturwissenschaften (2007) 94:967–974
amphibious-terrestrial Haemopidae/Hirudinidae (Trontelj
et al. 1999; Kutschera and Wirtz 2001).
Human-induced climate and habitat changes
Between 1960 and ca. 1971, Reisinger and the senior author
of this report (Ebermann) encountered a number of adult
Xerobdella individuals in the birch forests around Graz
(Fig. 1a), although it should be noted that this species has
always been rare. However, excursions to these localities
carried out between 2001 and 2005 yielded only one living
juvenile individual (Fig. 2), which suggests that this
population of land leeches has been sharply declining in
numbers. The climatologist Kabas (2005) compiled two
Austrian temperature records spanning the past four
decades (1961 to 2004). In two representative regions, the
national park Hohe Tauern and a flat area in the south eastern
part of the Steiermark, average air temperatures were
recorded and analysed in detail. These data document that
between 1961 and 2004, the mean annual air temperature
increased in these regions by 2.00 and 2.41°C, respectively.
In the southeastern part of the Steiermark, the increase in
average summer temperature was +3.05°C (Table 1). All
these recorded climate changes are statistically significant
(Kabas 2005). In addition, we recognized that the moist soil,
where Xerobdella individuals had been found until ca. 1971
under pieces of bark and rotten birch leaves, has become
much dryer. In the hot summer of 2003, some of the
original leech habitats around Graz had entirely dried up.
However, we have no quantitative data (soil moisture
contents) to further document these qualitative field
observations.
Discussion
In this study, we provide information on the morphology,
biology, feeding behaviour and DNA-taxonomy of X.
lecomtei, the only supposed member of the ‘land leech
family’that inhabits relatively cool birch forests of some
regions in Europe. Is this rare annelid in fact a close relative
of these aggressive, blood-sucking haemadipsids?
Ninety years ago, Blanchard (1917) published a review
article wherein he grouped together all blood-sucking leech
species with a terrestrial habit, inclusive of X. lecomtei, and
established the family Haemadipsidae. Most zoologists
followed him, notably Reisinger, who described the European
land leech as a relict taxon from the Tertiary (Moosbrugger
Fig. 3 Feeding behaviour of a
juvenile land leech (Xerobdella
lecomtei). The animal in knotted
posture (a) is alerted by an adult
earthworm (Lumbricus casta-
neus), rapidly attaches to its
prey and sucks body fluids (b).
Resting satiated leech (c)in
characteristic ball-like configu-
ration. Scale bar=10 mm
Fig. 4 Phylogenetic relation-
ships of ten selected leech spe-
cies with the Erpobdellidae as
outgroup. The diagram shows a
neighbour-joining tree obtained
from newly acquired and pub-
lished COI DNA sequence data,
inclusive of the branch support
values. Taxonomy of the leeches
is adapted from Kutschera and
Wirtz (2001)
Naturwissenschaften (2007) 94:967–974 971
and Reisinger 1971). This hypothesis has been accepted by
Sawyer (1986); Nesemann and Neubert (1999) and others. It
should be noted that Sawyer (1986) stressed that Xerobdella
has only four pairs of eyes (instead of five in typical
haemadipsids), but he nevertheless interpreted this European
taxon as a member of the tropical land leech family.
Although Reisinger’s hypothesis has been questioned (see
Soos 1966;Minelli1979), only with the advent of molecular
methods have new insights been obtained. Based on nuclear
18 S rDNA sequences, Trontelj et al. (1999) and other
investigators (Borda and Siddall 2004) found that Xerobdella
is more closely related to members of the genera Haemopis
and Hirudo than to tropical Haemadipsa species.
In this study, we use a DNA-barcoding approach and show
that on the basis of mitochondrial COI-sequence data, three
genera of haemadipsids from Asia, Australia and Madagascar
form a monophyletic clade, to the exclusion of the European
Xerobdella. It should be noted that the genera Haemadipsa
and Chtonobdella from Asia and Australia, respectively, are
well-characterized taxa (Richardson 1968; Bhatia 1975;
Sawyer 1986), but the biology and taxonomic status of
haemadipsids from Madagascar are unknown. In the most
recent monograph on the ‘Natural History of Madagascar’
(Goodman and Benstead 2003), leeches are not mentioned,
although these aggressive ectoparasites are well-known pests
to travellers (Bhatia 1975). In this study, we show that the
common species M. fallax (Richardson 1978), collected in
the centre of Madagascar for DNA extraction, is a close
relative of its Asian and Australian counterparts. Our
phylogenetic tree (Fig. 4) shows that Xerobdella is not a
member of the Haemadipsidae; it may be more closely
related to leeches of the genera Haemopis and Hirudo,as
suggested by Trontelj et al. (1999). However, the branch of
our tree leading to X. lecomtei has a very low support value;
that is, in our analysis, the position of the European land
leech towards other hirudineans is not conclusive. In
accordance with Minelli (1979), we assign the taxon X.
lecomtei to the family Xerobdellidae and to the suborder
Hirudiniformes (Nesemann and Neubert 1999).
The suggested Xerobdella–Haemopis relationship is
further corroborated by the facts that (1) both leech species
are terrestrial earthworm feeders that forage in a similar
way (Kutschera and Wirtz 2001), (2) they display an
‘earthworm-like’resting position during periods of diges-
tion (Sims and Gerard 1985;Fig.2a) and (3) are
characterized by a largely identical digestive tract and other
anatomical features (Moosbrugger and Reisinger 1971).
The similarities of Xerobdella to the recently discovered
terrestrial taxon H. elegans from Europe (Grosser 2004) are
especially striking (habitat preference and behaviour).
However, more work is required to further explore the
putative evolutionary relationship between members of the
genera Xerobdella and Haemopis (Trontelj et al. 1999). It
should be noted that our DNA-barcoding data indicate that
the common species H. sanguisuga and its rare, morpho-
logically similar relative H. elegans are different taxa, as
suggested on the basis of field observations and micro-
scopical studies (Grosser 2004).
In November 1867, Georg Ritter von Frauenfeld
submitted his classical paper on X. lecomtei for publication.
Only 140 years later, we have to describe the sad story of
‘Lonesome George of the annelids’(Nicholls 2006)—a
reminder that the impact of humans on the environment can
be more rapid and subtle than previously thought. The birch
forests around Graz still look like those where Reisinger
had collected his Xerobdella individuals (Moosbrugger and
Reisinger 1971). The data summarized in Table 1document
that a significant anthropogenic local warming had occurred
that is not restricted to the habitat of the leech population
investigated here. In the southeastern part of the Steiermark,
where Reisinger had found most of his specimens, an
increase in average summer temperature of +3.05°C is
documented for the time period between 1961 and 2004.
This local warming is in accordance with the global trend
reported in the literature (Hughes 2000; Travis 2002;
Schiermeier 2007). In the course of the hot summer of
2003, ‘tropical temperatures’(higher than 30°C) were
recorded on 41 days in the centre of Graz (Kabas 2005).
As a consequence of this drastic warming, a spread of the
European mantis (Mantis religiosa) occurred between 1970
and 1986 (Gepp and Kreissl 1988).This insect species
prefers dry, sunny meadows and can only reproduce in
warm regions.
The increase in air temperature led to a sharp decline in
the moisture content of the soil of the birch forests where
the land leeches live, notably in the hot summer of 2003
(Ebermann, unpublished observations). Because X. lecomtei
belongs to those specialist species that have a low
colonization ability and poor dispersal, this rare annelid
may be prone to extinction as a result of climate change
(Travis 2002). Reisinger reported that in the laboratory, the
optimal temperature for the cultivation and maintenance of
Xerobdella populations is around 12°C and that at 25°C
these cold-adapted worms rapidly die (Reisinger 1951;
Table 1 Mean increase in air temperature (°C) during the time period
1961–2004 in two regions of Austria
Region Altitude above
sea level (m)
Temperature increase (°C)
Hohe Tauern 500–3,106 2.00 (average), 2.34 (spring),
2.46 (summer)
Südoststeiermark 208–725 2.41 (average), 2.54 (spring),
3.05 (summer)
Data compiled from Kabas (2005).
972 Naturwissenschaften (2007) 94:967–974
Moosbrugger and Reisinger 1971). Based on our climate
data (Table 1) and field observations, we suggest that recent
human-induced warming may have led over past decades to
the almost complete extinction of a local population of this
rare animal species. It should be noted, however, that the
evidence for this hypothesis is only circumstantial. In
colder alpine regions of Austria, other Xerobdella popula-
tions may still exist in moist local refugia, but currently, we
have no proof for this assumption. Our ‘Lonesome George’
(Fig. 2) lives on as a COI GeneBank number (EF 125040)
that can now be used by other scientists for further
phylogenetic analyses (DNA barcoding initiative, see
Pfeiffer et al. 2004,2005; DeSalle et al. 2005; Bely and
Weisblat 2006; Waugh 2007).
The general conclusions and recommendations that
emerge from our study are threefold. First, human-induced
warming without apparent habitat destruction may lead to
subtle changes in biodiversity, notably the decline and
extinction of populations that consist of cold-adapted
species. Second, it is imperative to enhance research
budgets in systematic biology, biodiversity research and
taxonomy. We can much better protect organisms that
specialists have collected, classified and described in the
scientific literature than unknown biodiversity in regions
unexplored by ecologists (Miller 1999). Finally, museums
and biology departments should adequately preserve their
specimens at low temperatures for future generations of
scientists. The DNA-barcoding project (DeSalle et al. 2005;
Bely and Weisblat 2006; Waugh 2007; Kutschera 2007)
and other initiatives for the documentation and analysis of
global biodiversity will depend on collections of ethanol-
preserved specimens in this era of anthropogenic climate
change and habitat destruction (Wilson 2003).
Acknowledgements This paper is dedicated to the naturalist Georg
Ritter von Frauenfeld (1807–1873) on the occasion of his 200th
birthday. We thank Mr. C. Grosser (Leipzig, Germany) and Dr. D. Rinke
(Vogelpark Walsrode, Germany) for the provision of leech specimens.
Financial support was provided by the Bundesministerium für
Wirtschaft und Technologie (Förderkennzeichen 03 ESFHE 021).
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