J. Parasitol., 95(1), 2009, pp.

20–24
䉷 American Society of Parasitologists 2009

NEUROLOGICAL AND PHYSIOLOGICAL DISORDERS IN ARTEMIA HARBORING

MANIPULATIVE CESTODES
Marta I. Sánchez*, Frédéric Thomas, Marie-Jeanne Perrot-Minnot†, David G. Biron‡, Justine Bertrand-Michel§, and
Dorothée Missé
Equipe ‘‘Evolution des Systèmes Symbiotiques,’’ GEMI, UMR CNRS/IRD 2724, IRD, 911 Avenue Agropolis, BP 64501, 34394 Montpellier
CEDEX 5, France. e-mail: marta.sanchez@cefe.cnrs.fr

ABSTRACT: There are many impressive examples of host manipulation by parasites, but mechanisms underlying these ethological
changes, as well as their physiological consequences, are not well characterized. Here, we analyzed part of the cerebral proteome
of brine shrimp Artemia infected by manipulative cestodes, using for the first time the ProteinChip Surface-Enhanced Laser
Desorption Ionization and Time of Fly Mass Spectrometry (SELDI-TOF MS) system, which has been proposed as an excellent
way to analyze the host genome during the host–parasite interaction processes. We found 2 peptides downregulated in individuals
infected by the dilepidid, Anomotaenia tringae (4.5 kDa), and by the 2 hymenolepidids, Flamingolepis liguloides and Confluaria
podicipina (3.9 kDa), which are potential candidates for involvement with the manipulation process. The identification of 2 head
peptides (4.1 and 4.2 kDa) overexpressed in all the categories in brine shrimp living at the surface (both infected individuals and
uninfected controls) suggests its association with the different environmental conditions experienced at the water surface. In
parallel, brine shrimp infected by C. podicipina showed significant values of triglycerides, potentially augmenting their profit-
ability and attractiveness for the predaceous definitive host (grebes). We discuss our findings in relationship with current ideas
on the complexity of parasitically modified organisms.

During the last 3 decades, the study of phenotypic changes
in parasitized organisms has attracted the interest of many re-
searchers (Adamo, 2002; Moore, 2002; Thomas et al., 2005).
In many of the best-known examples, changes displayed by
infected hosts appear advantageous for the parasites. For in-
stance, parasite-induced alterations in host behavior can result
in the infected host becoming more vulnerable to predation by
the parasite’s next host (Lafferty, 1999; Poulin, 2007). The al-
tered host phenotype may also cause the host to migrate toward
particular microhabitats, where the parasite must be to complete
its transmission cycle (Moore, 2002; Poulin, 2007). An impor-
tant aspect of recent articles on parasite manipulations (see
Thomas et al., 2005) is the recognition that these phenomena
are probably more complex than traditionally viewed, i.e., ma-
nipulated hosts being altered by a range of changes, in addition
to the most obvious ones, e.g., aberrant behavior. Along the
same idea, it seems that the mechanisms underlying ethological
changes following infection are in many cases sophisticated,
some sort of chemical interference with host neurochemistry,
endocrine function, and/or gene expression (Thomas et al.,
2005).
The representatives of the cosmopolitan crustacean Artemia
(Branchiopoda: Anostraca) are key species in extreme hyper-
saline environments (e.g., Sánchez, Green et al., 2006). In salt-
pans of southwest Spain, Artemia parthenogenetica is inter-
mediate host for at least 10 species of cyclophyllidean tape-
worms, the adults of which infect several species of water birds
(see Georgiev et al., 2005). Brine shrimp become infected by
consuming cestode eggs released into the water with the feces
Received 6 December 2007; revised 9 April 2008, 26 May 2008;
accepted 26 May 2008.
* Present address: CEFE, UMR 5175, CNRS, 1919 Route de Mende,
34293 Montpellier, France.
† Equipe Ecologie Evolutive, UMR CNRS 5561 Biogeosciences, Uni-
versité de Bourgogne, 6 Boulevard Gabriel, 21000 Dijon, France.
‡ HPMCT Proteome Analysis, 223 St. Antoine, St. Elphège, Quebec,
Canada J0G 1J0.
§ INSERM, Institut Claude de Préval, IFR30, Plateau Technique de Lip-
idomique, Toulouse F-31300, France.
of the avian definitive hosts. The newly hatched oncosphere
penetrates into the hemocoel, where it develops into a cysticer-
coid. In at least 3 cestode species (Flamingolepis liguloides
infecting flamingos, Confluaria podicipina infecting grebes, and
Anomotaenia tringae infecting shore birds), the presence of cys-
ticercoids is associated with changes in host behavior and color
(Gabrion et al., 1982; Amat et al., 1991; Robert and Gabrion,
1991; Sánchez, Georgiev et al., 2006; Sanchez et al., 2007),
making infected individuals more vulnerable to foraging birds.
Adult tapeworms develop and reproduce in the small intestine
of bird final hosts, concluding their life cycle.
We first designed the present study to explore some of the
proximate mechanisms underlying behavioral modifications in
parasitized A. parthenogenetica. Although it is generally ad-
mitted that manipulative parasites can change host behavior by
directly and/or indirectly modulating the host’s central nervous
system, empirical demonstrations of such neurophysiological
interaction between manipulated hosts and parasites are very
scarce. Recently, proteomics has been shown to offer an ex-
cellent way to analyze the host genome during the host–parasite
interaction processes (Biron, Joly et al., 2005). Here, we used
for the first time The ProteinChip Surface-Enhanced Laser De-
sorption Ionization and Time of Fly Mass Spectrometry (SEL-
DI-TOF MS) system (Cipergen娂, Freemont, California) (Vor-
derwülbecke et al., 2005), to contrast the head proteome of
uninfected individuals with brine shrimp infected by several
species of manipulative cestodes. Because of parasite-induced
behavioral changes, we predict that there should be substantial
differences in the protein profiles of infected and uninfected
individuals. A second aim of this work was to compare the lipid
contents of infected and uninfected A. parthenogenetica. In-
deed, for reasons that are still unclear, several recent articles
suggest that aquatic crustaceans parasitized by manipulative
helminths completing their life cycles in aquatic birds, i.e.,
trematodes (Ponton et al., 2005), acanthocephalans (Plaistow et
al., 2001), and cestodes (Amat et al., 1991), possess signifi-
cantly higher levels of lipids than uninfected conspecifics. Here,
we attempted to further probe several components of the lipid
storage forms in infected and uninfected A. parthenogenetica.

20
 SÁNCHEZ ET AL.—NEUROPHYSIOLOGICAL DISORDERS IN ARTEMIA ARBORING CESTODES
MATERIALS AND METHODS
Sampling
Artemia parthenogenetica were collected with a 0.1-mm-mesh net
from an industrial saltpan in the Odiel Marshes (Huelva, southwest
Spain, 37⬚17⬘N, 06⬚55⬘W) during August 2005 for the proteomic anal-
ysis and June 2006 for the lipid analysis. Infected individuals displaying
aberrant behavior were sampled from the water surface, whereas unin-
fected ones were collected from the bottom (a direct consequence of
the infection is the spatial segregation of infected and uninfected indi-
viduals in the water column). Given the important ecological differences
between these 2 microhabitats (food, light, oxygen, disturbance, tem-
perature, predation risk, etc.), and knowing that not only infection sta-
tus, but also environmental conditions, may have a significant influence
on protein expression, we created a control group by placing uninfected
brine shrimp from the bottom in the same ecological conditions as in-
fected ones from the surface (as a control for environmental conditions).
We placed a sample of uninfected brine shrimp in a metallic cage (50-
cm diameter) located at the surface (1 cm) at the collecting site for 24
hr. For the host head proteome analysis, we used A. parthenogenetica
as host species and 3 cestode species, i.e., F. liguloides, C. podicipina,
and A. tringae, known to use brine shrimp as intermediate hosts. For
the SELDI-TOF experiment, we used 5 categories of individuals: (1)
uninfected A. parthenogenetica from the bottom (NP); (2) uninfected
A. parthenogenetica from the bottom placed at the surface as controls
(NPC); (3) A. parthenogenetica infected by F. liguloides (AP FL); (4)
A. parthenogenetica infected with C. podicipina (AP CP); and (5) A.
parthenogenetica infected by A. tringae (AP AT). For the lipid analysis,
we were unable to find enough individuals infected by A. tringae so
that only 3 categories were considered, i.e., uninfected individuals, in-
fected by F. liguloides, and infected by C. podicipina.
In the laboratory, immediately following the collection, individuals
were carefully checked for the presence of cysticercoids, dried on ab-
sorbent paper, and individually stored in a 1.5-ml Eppendorf tube at
⫺80 C until the analyses were undertaken. Only individuals infected
with 1 cestode species were employed in the study. Using a partheno-
genetic species (all the individuals were females) limits possible host
sex-specific effects. For the SELDI-TOF analysis, only the heads, after
being carefully removed from the bodies, were preserved on ice. The
bodies were then placed in temporary glycerol mounts and cysticercoids
were examined with the use of stereo or ordinary light microscopy to
identify the cestode species. If the identification of the cysticercoids
was not possible at this stage, whole infected brine shrimp or isolated
cysticercoids were prepared as permanent mounts in Berlese’s medium
to facilitate observations on the morphology of rostellar hooks. The
same protocol was followed to verify the absence of cysticercoids for
individuals collected from the bottom and those experimentally placed
in the surface.
Proteomic analysis (SELDI-TOF MS)
The ProteinChip SELDI system was used as previously described
(Missé et al., 2007). We used protein chip arrays, i.e., solid-phase chro-
matographic surface chips, to ionize proteins from host heads and then
analyze them. Preactivated differential binding surfaces permitted mul-
tidimensional chromatography. Three types of protein chip surfaces, i.e.,
hydrophobic (H50), normal phase (NP20), and immobilized metal af-
finity capture (IMAC 30), were used to perform preliminary analysis to
search for the most suitable binding surface for protein retention. IMAC
30 ProteinChip Arrays (Ciphergen Biosystems, Fremont, California)
previously activated with 50 mmol/L NiSO4 (according to manufactur-
er’s instructions) were used to perform the final analysis.
In total, the heads of 52 uninfected, 42 uninfected controls, and 56
infected brine shrimp were examined. Infected A. parthenogenetica in-
cluded 21 by A. tringae, 11 by F. liguloides, and 24 by C. podicipina.
Brine shrimp heads were individually bound to activated chip spots and
incubated for 24 hr at 4 C. After incubation, unbounded proteins were
removed by 3 successive washes of phosphate-buffered saline. Chip-
captured proteins were air dried at room temperature for 15 min, fol-
lowed by 2 additions (1 ␮l each) of a saturated solution of ␣-cyano-4-
OH-cinnamic acid (CHCA) matrix. The ionized and desorbed proteins
were detected and their molecular masses displayed on a proteogram.
Protein peaks were analyzed by SELDI-TOF MS analysis with the Pro-
tein-Chip Biology System II software (PBS II) and Ciphergen Peaks
21
software (Ciphergen Biosystems). A range of 2–30-kDa molecular
weights (SELDI TOF-MS is especially sensitive at this low-molecular-
mass range) was analyzed for differences in peptide profile. Spectra
were mass aligned, baseline-subtracted with the use of a smoothing
feature, and then normalized with the use of total ion current normali-
zation before statistical analysis was performed. The total ion current
method assumes that, on average, the total number of proteins being
expressed is constant across the samples being normalized. The process
takes the total ion current used for all the spots, averages the intensity,
and adjusts the intensity scales for all the spots to display data on the
same scale. The m/z of each peak to be quantified was determined
according to externally calibrated standards (Ciphergen Biosystems).
Univariate analyses, Mann–Whitney nonparametric U-tests, were used
to compare the mean intensity values of each recognized peak at the
molecular mass range of 2–30 kDa. A significance threshold was set at
P ⬍ 0.05.
Lipid analysis
We measured free cholesterol, diglycerides (including phospholipids,
DG ⫹ PL), cholesterol esters (Cho Est), and triglycerides (TG). The
measurements of neutral lipid molecular species were performed by
gas–liquid chromatography. Brine shrimp (10 individuals per group)
were individually crushed with the use of an Ultra Turax in 1 ml of
methanol/5 mM EGTA (2:1 v/v). Aliquots (50 ␮l) were evaporated, the
dry pellets were dissolved in 0.2 ml of NaOH (0.1 M) overnight, and
proteins were measured with the Bio-Rad assay (Bio-Rad Laboratories,
Hercules, California). Lipids corresponding to 0.5 ml of the homoge-
nized body were extracted according to Bligh and Dyer (1959) in chlo-
roform/methanol/water (2.5:2.5:2.1, v/v/v), in the presence of the inter-
nal standards: 6 ␮g of stigmasterol, 2 ␮g of 1,3-dimyristine, 2 ␮g of
cholesteryl heptadecanoate, and 3 ␮g of glyceryl triheptadecanoate. The
chloroform phase was filtered over glass wool, evaporated to dryness,
and dissolved in 20 ␮l of ethyl acetate. One microliter of the lipid
extract was analyzed by gas–liquid chromatography on a FOCUS Ther-
mo Electron system (Waltham, Massachusetts) with the use of a Zebron-
1 Phenomenex (Phenomenex, Torrance, California) with fused silica
capillary columns (5 m ⫻ 0.32–mm i.d., 0.50-mm film thickness) (Bar-
rans et al., 1994). Oven temperature was programmed from 200 to 350
C at a rate of 5 C per min and the carrier gas was hydrogen (0.5 bar).
The injector and the detector were at 315 and 345 C, respectively.
All statistical analyses were performed following Sokal and Rohlf
(1981), Siegel and Castellan (1988), and Zar (1999). When data devi-
ated from normality and/or did not fit a normal distribution after trans-
formation, we used nonparametric statistics. Post hoc comparisons were
performed according to Zar (1999) with a significance level of P ⬍
0.05. P values were Bonferroni corrected to avoid Type I errors.
Throughout the article, values given are mean ⫾ SE. Results were con-
sidered significant at the 5% level.
RESULTS
Proteomic analysis
SELDI-TOF analysis of A. parthenogenetica head extracts
showed specific peptide profiles for each category in the range
of 2–5 kDa (Fig. 1). This representative peptide profile also
revealed several differentially expressed peptide peaks. The
presence of a soluble head peptide with a molecular mass of
4.5 kDa was underexpressed in the heads of A. tringae–infected
individuals (AP AT) compared with controls (NPC and NP), as
well as in the other 2 infected groups (AP CP and AP FL; P
⬍ 0.001). SELDI analysis also showed an absence of a 3.9-
kDa head peptide in A. tringae- and F. liguloides–infected in-
dividuals compared with controls and C. podicipina–infected
individuals. Two head peptides of 4.1 and 4.2 kDa were over-
expressed in the majority of infected individuals, as well as in
uninfected controls placed at the surface (P ⬍ 0.05).
22 THE JOURNAL OF PARASITOLOGY, VOL. 95, NO. 1, FEBRUARY 2009
FIGURE 1. Surface-enhanced laser adsorption/ionization time-of-
flight (SELDI-TOF) MS analysis of head proteome of Artemia showing
differentially expressed protein peaks. NPC (nonparasitized control
placed at the surface), NP (nonparasitized), AP AT (infected by An-
omotaenia tringae), AP CP (infected by Confluaria podicipina), AP FL
(infected by Flamingolepis liguloides). Arrows indicate peptidic bio-
marker.
Lipid analysis
It was not possible to select individuals of the same size for
infected and uninfected categories, because some species, e.g.,
F. liguloides (Amat et al., 1991), induce gigantism. Thus, the
mean body size was slightly different between uninfected
(10.45 ⫾ 0.33, mean ⫾ SE) and infected individuals (11.4 ⫾
0.33 for F. liguloides; 11.9 ⫾ 0.34 for C. podicipina) (ANOVA,
F ⫽ 4.96, P ⫽ 0.015; post hoc comparisons, Student’s t-test, P
⬍ 0.047, for uninfected–F. liguloides and uninfected–C. podi-
cipina). To avoid size-confounding effects, we performed all
the analyses on the residuals of the relationship between lipid
content (ln) and brine shrimp size (ln). Details of lipid content
for the different categories are presented in Table I. Values for
diglycerides were in general significantly higher in uninfected
A. parthenogenetica than in infected ones. Conversely, for tri-
glycerides and Cho Est content, the general tendency was to
exhibit an increase from individuals infected with F. liguloides
(with minimum values), followed by uninfected controls, to
those infected with C. podicipina (with the highest values; Ta-
ble I). Differences between uninfected individuals and those
infected with C. podicipina were significant for TG C49. There
was no significant effect of parasite load on the lipid content
(Spearman rank test, P ⬎ 0.107 for C. podicipina and P ⬎ 0.08
for F. liguloides).
DISCUSSION
Proteomic analysis
Studying processes that explain parasite strategies is critical
to understanding the evolution of host–parasite interactions. Al-
teration of the central nervous system is commonly used by
parasites to alter host behavior (Holmes and Zohar, 1990;
Thompson and Kavaliers, 1994; Adamo, 2002; Helluy and
Thomas, 2003; Beckage and Gelman, 2004; Biron, Marché et
al., 2005), although the mechanisms responsible are not well
characterized.
Proteomics constitute a powerful tool to investigate the mo-
lecular cross talk established during the manipulative processes
(Biron, Moura et al., 2005; Thomas et al., 2005). A major ap-
proach for protein biomarker discovery is differential protein
expression profiling, in which protein expression levels are
compared across samples. Here we used for the first time SEL-
DI-TOF MS to compare the head proteomes of uninfected or-
ganisms and those infected by a manipulative parasite. Al-
though this approach permitted a rapid discovery of differen-
tially expressed proteins in manipulated individuals, difficulty
in identifying proteins of interest is a major limitation of this
technology. Despite this problem, a SELDI-TOF approach per-
mitted rapid and reproducible protein profiling directly from
crude samples, and allowed us to bind a range of proteins and
peptides to different molecular surfaces without the requirement
for antibody production.
In the present study, SELDI-TOF analysis detected 2 peptides
whose expression is altered in individuals infected by the di-
lepidid, A. tringae (downregulated 4.5-kDa peptide), and by the
2 hymenolepidids F. liguloides and C. podicipina (downregu-
lated 3.9-kDa peptide). Because parasitized individuals also
typically displayed the altered behavior, these peptides are po-
tential candidates for involvement with the manipulation pro-
cess. However, to confirm this hypothesis, further work is re-
quired, particularly in identification of these molecules and their
biological functions.
Another interesting finding is the identification of 2 head pep-
tides (4.1 and 4.2 kDa) overexpressed in all the categories in
brine shrimp living at the surface (both infected individuals and
uninfected controls). This suggests that some peptide expres-
sions are more the consequence than the cause of the behavioral
change, i.e., different environmental conditions experienced at
the water surface. This of course also underlines the need in
proteomic investigations for considering control categories.
Further identification of these peptides would be necessary to
understand their role. At the moment, we can speculate about
their relation in the synthesis of all-trans-canthaxanthins, as in-
fected photophilic individuals (displaying bright red coloration)
are more exposed to UV radiation, which stimulates synthesis
of photoprotective carotenoids. The next step to improve our
understanding of the molecular mechanisms implied in the
modification of host behavior in this system will be the iden-
tification (isolation, purification, and characterization) of the
 SÁNCHEZ ET AL.—NEUROPHYSIOLOGICAL DISORDERS IN ARTEMIA ARBORING CESTODES 23

TABLE I. Artemia parthenogenetica lipid content (nmol/mg, mean ⫾ SE) for individuals infected with Confluaria podicipina (cp), Flamingolepis
liguloides (fl), and for uninfected individuals (ni). Measures of diglycerides (DG) include phospholipids (PL).

Post hoc comparisons

ANOVA (Bonferroni corrected)
Composes Uninfected C. podicipina F. liguloides F P ⬍0.05 ⬍0.01

Cholesterol 27.96 ⫾ 3.89 22.99 ⫾ 2.45 28.19 ⫾ 4.71

DG 16-16 0.00 ⫾ 0.00 0.00 ⫾ 0.00 0.88 ⫾ 1.29
DG 16-18 21.96 ⫾ 6.40 8.82 ⫾ 2.30 10.82 ⫾ 2.34 10.25 * ni ⬎ cp, fl
DG 18-18 22.79 ⫾ 7.66 9.90 ⫾ 3.49 14.20 ⫾ 4.90
DG 18-20 0.00 ⫾ 0.00 0.00 ⫾ 0.00 0.78 ⫾ 1.68
DG ⫹ PL 44.75 ⫾ 13.60 18.72 ⫾ 5.27 26.67 ⫾ 7.99 ni ⬎ cp
C18 0.55 ⫾ 0.49 0.06 ⫾ 0.19 0.04 ⫾ 0.08
C20:4 2.96 ⫾ 1.94 6.35 ⫾ 2.97 2.33 ⫾ 2.52
Total cholesterol esters 3.65 ⫾ 1.48 6.41 ⫾ 2.86 2.37 ⫾ 2.48 7.86 † cp ⬎ fl
Triglycerides (TG) C49 0.50 ⫾ 0.82 3.89 ⫾ 1.92 2.10 ⫾ 2.29 cp ⬎ ni
C51 12.79 ⫾ 6.82 17.99 ⫾ 8.18 8.21 ⫾ 9.00
C53 27.14 ⫾ 12.50 38.35 ⫾ 18.74 15.89 ⫾ 18.06 7.65 † cp ⬎ fl
C55 27.25 ⫾ 13.87 42.39 ⫾ 25.40 16.14 ⫾ 19.91 8.19 † cp ⬎ fl
C57 16.15 ⫾ 8.67 27.86 ⫾ 17.93 9.56 ⫾ 11.46 8.87 † cp ⬎ fl
Total TG 83.84 ⫾ 39.80 130.49 ⫾ 68.58 51.90 ⫾ 60.01 7.88 † cp ⬎ fl
Pic 1 (27,85⬘) 13.42 ⫾ 6.50 19.33 ⫾ 11.15 7.43 ⫾ 8.02 cp ⬎ fl
Pic 2 (29,27⬘) 4.60 ⫾ 2.56 14.85 ⫾ 14.34 3.25 ⫾ 3.69 cp ⬎ fl
Pic 3 (30,59⬘) 3.01 ⫾ 1.59 5.08 ⫾ 3.37 1.64 ⫾ 1.86 cp ⬎ fl

* P ⬍ 0.01 after Bonferroni correction.

† P ⬍ 0.05 after Bonferroni correction.

candidate peptides potentially linked to the manipulative pro-
cess, followed by functional analysis.
Effect of infection on lipid levels
Contrary to the findings of Amat et al. (1991) showing higher
levels of total lipid reserves in F. liguloides–infected brine
shrimp, we found an opposite result. Thus, there were decreased
levels of all the specific lipids analyzed. However, brine shrimp
infected by C. podicipina (adult parasite of Black-necked grebe
Podiceps nigricollis) showed a tendency (significant for TG
C49) to increased levels of triglycerides. By increasing the en-
ergetic level of their intermediate host, C. podicipina potentially
augment their profitability and attractiveness for the grebe final
host. Fatty acids in wild bird food are mostly in the triglyceride
form (Zurovchak et al., 1999), with elevated plasma of triglyc-
eride levels in birds being indicative of fat deposition (Robin-
son, 1970; Ramenofsky, 1990).
Black-necked grebes forage primarily on brine shrimp, a
highly digestible and energy-rich food for them (Caudell and
Conover, 2006). The Odiel marshes (in southwestern Spain) are
1 of the major stopover sites for grebes when they molt. This
period, in which grebes become flightless for up to several
months, is a very energy-demanding experience; there are dra-
matic increases in body fat stores, leading to a doubling of their
body mass (Jehl, 1997). Observations of grebes picking brine
shrimp from the surface (infected and segregated individuals)
instead of the most common way to feed, i.e., while diving (M.
I. Sánchez, pers. obs.), support the idea that infected A. par-
thenogenetica, with nearly double the normal levels of triglyc-
erides, are the preferred prey during this period of time.
Alternative explanations to the ‘‘signaling hypothesis’’ can
also be proposed. For example, lipid content has been proposed
to be a good predictor of copepod survival (Franz and Kurtz,
2002), so that manipulation of energetic storages may also func-
tion as a parasite strategy to increase the longevity of infected
individuals and, therefore, their probability of encountering a
final host. The greater survival of infected crustaceans reported
by Amat et al. (1991) supports this hypothesis.
Artemia parthenogenetica is a keystone species in hypersa-
line habitats and is a major food resource not only for grebes,
but also for a wide range of water birds (Cooper et al., 1984;
Britton et al., 1986; Verkuil et al., 2003; Sánchez et al., 2005),
especially during migration, when it is well established that
birds accumulate fat reserves by increasing food intake and by
choosing high-energy diets (Blem, 1990; Stiles, 1993; Bairlein
and Gwinner, 1994; Biebach, 1996; McWilliams and Karasov,
2004). Because the cestodes studied here appear to enhance not
only the accessibility to predation, but, in the case of species
such a C. podicipina, also their nutritional value (specific tri-
glycerides), ecologists and conservationists should devote fur-
ther attention to exploring the relationships between parasites
and the trophic potential of habitats.
ACKNOWLEDGMENTS
The first author was supported by a postdoctoral grant from the Min-
isterio de Ciencia y Tecnologı́a (Spain). Consejerı́a de Medio Ambiente,
Junta de Andalucı́a, and Aragonesas Industrias y Energı́a S.A. provided
permission to work in the saltworks. We are grateful to A. J. Green,
anonymous referees, the Associate Editor Mike Sukhdeo, and the Editor
Gerald W. Esch for helpful comments on the manuscript.
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