Journal Pre-Proof: Science of The Total Environment
Journal Pre-Proof: Science of The Total Environment
Journal Pre-Proof: Science of The Total Environment
PII: S0048-9697(21)04998-6
DOI: https://doi.org/10.1016/j.scitotenv.2021.149923
Reference: STOTEN 149923
Please cite this article as: J. Boutry, J. Mistral, L. Berlioz, et al., Tumors (re)shape biotic
interactions within ecosystems: Experimental evidence from the freshwater cnidarian
Hydra, Science of the Total Environment (2018), https://doi.org/10.1016/
j.scitotenv.2021.149923
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1-CREEC/CANECEV (CREES), MIVEGEC, Unité Mixte de Recherches, IRD 224–
CNRS 5290–Université de Montpellier, Montpellier, France
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2-Zoological Institute, Christian-Albrechts University, Kiel, Germany
3-MTA-DE Behavioural Ecology Research Group, Department of Evolutionary Zoology,
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University of Debrecen, 4032 Debrecen, Hungary
4-AZELEAD, 377 Rue du Professeur Blayac, 34080 Montpellier, France
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5-Centre for Integrative Ecology, School of Life and Environmental Sciences, Deakin
University, Waurn Ponds, Victoria, Australia
6-LIENSs, UMR 7266 CNRS-La Rochelle Université, 2 Rue Olympe de Gouges, 17000
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La Rochelle, France
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AUTHORSHIPFLY,
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JB, MG, BU and FT designed the study. JB, JM, LB and LF collected the data, while JB,
JT and AD performed the analyses. JT and AK provided biological material and technical
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training. JB wrote the manuscript, and all authors significantly contributed to revisions.
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ABSTRACT
While it is often assumed that oncogenic processes in metazoans can influence species
interactions, empirical evidence is lacking. Here, we use the cnidarian Hydra oligactis to
experimentally explore the consequences of tumor associated phenotypic alterations for its
Unexpectedly, hydra’s predation ability was higher in tumorous polyps compared to non-
tumorous ones. Commensal ciliates colonized preferentially tumorous hydras than non-
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tumorous ones, and had a higher replication rate on the former. Finally, in a choice
experiment, tumorous hydras were preferentially eaten by a fish predator. This study, for the
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first time, provides evidence that neoplastic growth has the potential, through effect(s) on
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host phenotype, to alter biotic interactions within ecosystems and should thus be taken into
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account by ecologists.
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Over the last few decades, great attention has been devoted by ecologists to
understanding the relationships between biological community dynamics and the functioning
of ecosystems (Loreau, 2010; Massol et al., 2011; Petchey and Gaston, 2002; Thompson et
al., 2012). It is now clearly established that the outcome of species interactions and their
ecosystem consequences are often strongly influenced by variation in the functional traits of
the organisms (Chalmandrier et al., 2017; Kishida et al., 2010; Raffard et al., 2019; Wright et
al., 2016). Factors influencing individual phenotypic variability are numerous, including
genetic and epigenetic ones in interaction with external biotic and abiotic variables (e.g.,
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predation, food availability (Chalmandrier et al., 2017; Raffard et al., 2019; Reed et al.,
2010)), as well as internal biotic factors associated with the holobiont. For instance, host-
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microbe interactions are known to influence animal behavior and life-history traits (Alcock et
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al., 2014; Walters et al., 2020; Wong et al., 2015). Besides, many manipulative parasites have
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the capacity to alter a broad range of phenotypic traits of their hosts, from color to
morphological traits and behavior (Hughes et al., 2012; Hughes and Libersat, 2019; Moore,
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2013; Thomas et al., 2011). Because alterations in the phenotype of parasitized hosts can be
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substantial, it has been argued that manipulated hosts can be ecologically equivalent to new
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organisms in the ecosystem, involved in novel direct and/or indirect interactions with other
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species (see also Mouritsen and Poulin, 2010; Pascal et al., 2020; Thomas et al., 1998).
another category of living entity inside their body: the community of neoplastic cells (Aktipis
and Nesse, 2013; Thomas et al., 2017). Neoplastic cell transformation affects most, if not all,
multicellular organisms, from hydras to whales (Aktipis et al., 2015; Albuquerque et al.,
2018; Madsen et al., 2017). Prior to being, in some cases, fatal for their hosts, tumors often
results in the alteration of phenotypic traits in their hosts, e.g. in morphology (Barr et al.,
2016; Jones et al., 2016), physiology (Argilés et al., 2014), body odors (Buszewski et al.,
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2012), reproductive activities (Arnal et al., 2017), as well as social interactions (Dawson et
al., 2018; Hamilton et al., 2020). Despite this large range of phenotypic alterations and the
processes on ecosystems and animals’ life histories remain largely unexplored (Giraudeau et
al., 2018).
microcosm, which allows empirically testing of whether bearing tumors may have
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implications for biotic interactions within ecosystems. The freshwater cnidarian Hydra
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oligactis (further referred to as hydra) is naturally found in aquatic habitats attached to
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aquatic vegetation or submerged rocks. Hydras reproduce asexually by budding but can also
invertebrates and vertebrates. For instance, they are predators of various zooplankton species
or even fish spawn (Elliott et al., 1997; Rivera-De la Parra et al., 2016). They can also serve
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as host for commensal (Coleman, 1966) or parasite (Stiven, 1965) species. Finally, they can
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be a prey for fish or mollusks grazing on plants (Baumga and Constance, 2005; Cuker and
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Mozley, 1981a).
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oligactis polyps can spontaneously develop tumors. These neoplasia not only severely alter
the polyp’s body shape, but tumor-bearing individuals also show a shift in their microbiota
and have a higher number of tentacles (Fig1, see also Domazet-Lošo et al., 2014; Rathje et
al., 2020). These tumors reduce the hosts’ fitness but rarely kill them, and are vertically
transmitted to the buds when polyps reproduce asexually, resulting into the spread of this
neoplasia into this lineage. Conversely offspring resulting from sexual reproduction are
tumor free (Domazet-Lošo et al., 2014). The prevalence of tumor-bearing individuals in the
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natural environment is unknown, but the observation of tumorous hydras in strains derived
from wild sampled individuals suggests that hydras in natural populations can be susceptible
to tumor development (Figure 1). These tumors appear after 3 to 4 months of maintenance in
stable laboratory conditions and seem to be composed of aberrant germinal cells, like the
tumors already described. However, more work is required to understand the causes of
Given the substantial phenotypic differences between healthy and tumorous hydras
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(Figure 2), we hypothesized that tumorigenesis might influence the biotic relationships
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involving hydras. To test this hypothesis, we experimentally tested for the first time the
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consequences of tumor associated phenotypic alterations on three types of biotic interactions:
hydras’ ability to catch prey, their capacity to serve as a host for a commensal ciliate and their
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vulnerability to predators. Through this pioneer study, we aim to improve our understanding
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Clonal non-tumorous and tumorous individuals of Hydra oligactis strains (St. Petersburg
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strains as control and tumorous lineages from the laboratory of Thomas Bosch1) were used
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for all experiments (Domazet-Lošo et al., 2014). Polyps were maintained at 18°C in Volvic©
water and fed three times per week with freshly hatched Artemia salina nauplii according to
standard protocols in tanks of 500mL (Lenhoff and Brown, 1970). Artemia nauplii were
obtained by adding 0.5g of eggs microcyst (Artemia salina, Planktovie S.A.S., Marseille,
France) in 400ml of seawater prepared with 36 g/L of sea salt (Reef Crystals, Aquarium
systems, Sarrebourg, France) and oxygenated with an aquarium pump. After 24 to 48 hours
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Zoological Institute, Christian-Albrechts-University Kiel, Am Botanischen Garten 1–9, D-24118 Kiel,
Germany.
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of incubation at 30°C, nauplii hatched and were collected with a pipette, rinsed with a filter
and suspended in a 200mL beaker of Volvic© water. All the experiments took place in
For each experiment, one hydra was placed in the experimental tank (2 mL well of a 6 well-
plate (Thermo Scientific) and left for two minutes of habituation to allow the polyp to
reattach to the substrate. Prey were added at two different density levels: In ad libitum
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feeding (by adding 250 µL taken from the bottom of the Artemia beaker, representing more
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than hundred nauplii) or restricted feeding (by adding 250 µL of Volvic from the upper part
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of the beaker containing around 10 nauplii in total). The total number of nauplii added in
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restricted feeding was re-counted at each experiment (since some individuals can sometimes
remain attached to the pipette tips, or conversely additional individuals can be introduced)
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and variations were taken into account in the analysis. In each experimental group (non-
ad libitum) 42 hydras were measured. One tumoral hydra of the restricted feeding condition
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was excluded because of an error in the nauplii number introduced. The predation capacity of
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hydra was calculated by counting the number of preys captured in their tentacles in 10
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The ciliates Kerona pediculus (named ciliate thereafter) are naturally present on hydra both in
the lab and in the wild, and considered as commensals (Coleman, 1966) and hence were used
polyps from an H. oligactis C2/7 strain previously sampled in Hungary and maintained in J.
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Tökölyi's laboratory2. Ciliates were taken from C2/7 strain hydra using a one-milliliter pipette
tip, under a binocular magnifier, by mildly aspirating the water around the hydra. In a first
experiment (free-choice), we introduced one ciliate into the 1.5 mL well of a microplate (12
well-plate, Thermo Scientific) containing one tumorous and one non-tumorous hydra (both
free from ciliates). After 4 hours, we recorded on which hydra the ciliate was present, this
ciliate into a 1.5 mL well containing one hydra, tumorous or non-tumorous. We first verified
the presence of the ciliate 15h after the inoculation, before the ciliate started to asexually
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reproduce. The number of ciliates on each individual was counted daily over six days on 36
non-tumoral and 36 tumoral individuals. The trials for which ciliates failed to colonize the
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host were removed to focus the analysis on colonization dynamics.
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(c) Predation risk on hydra
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To model the predation interactions, we used Siamese Fighting fish (Betta splendens) as
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predator. In the aquarist circle, some fishes are well-known to consume Hydra efficiently
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(Sharpe, 2020) but it is poorly documented in the literature (Baumga and Constance, 2005;
Cuker and Mozley, 1981b). Adult females were obtained from pet shops (Oxyfish©
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Verlinghem and Botanic©, Clapiers, France). Each fish was introduced into 500ml plastic
tanks (16x8.4x6.5cm) in Volvic© water at 25°C at least one hour before the test. A tumorous
and a non-tumorous polyp were introduced together in the central well (5 mL) of a standard
six well plastic plate for at least ten minutes of acclimation. The plate containing the two
polyps was then introduced progressively in the container with the fish and pressed the
bottom against the wall in front of the camera to allow recording and visualization of each
hydra during the test. The open part of the plate faced the tank to let the fish consume the
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Department of Evolutionary Zoology and Human Biology, University of Debrecen, Hungary.
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polyps attached in the central well of the plate. Two observers identified precisely the
position of each hydra during the test to recognize which one was consumed at each
predation event. The test stopped after the predation of the two hydras or after 45 minutes in
any case (i.e., no predation or only one specimen predated). This test was replicate 51 times
Statistical analysis
The effect of the tumorous phenotype on hydra predation was assessed by comparing the
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total number of Artemia nauplii captured by the two types of hydra (tumorous and non-
tumorous). For each model we started by representing the distribution of the variable to
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identify the relevant law to use, the normality was tested with Shapiro tests. When the
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variable was a proportion, we use binomial laws and when it was a count data we used
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Poisson law, negative binomial in case of overdispersion with a Poisson model or to model
the number of failures before a success. The selection of random and fixed effects to include
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in the analyses was made by model comparison (see Royle et al., 2013). The quality of
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models was evaluated by the package DARHMa, testing the homogeneity of the distribution,
the distribution of the residuals, the overdispersion, the presence of deviant outlier and the
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quantile deviation.
restricted feeding condition, we used a binomial glmm taking into account the proportion of
artemias consumed over the number introduced initially since small variation can exist
between trials. In both analyses, the hydra type was a fixed effect. Random effects of the date
was also included in ad libitum condition (see model choice in supplementary material). The
number of tentacles per polyp of each group was compared using a Wilcoxon test.
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The effect of the supernumerary tentacles of the tumorous phenotype on hydra predation
was assessed by comparing the total number of nauplii captured reported to the total number
of tentacles. In ad libitum condition, we used a log-normal linear mixed model with the
tumorous status as a fixed factor and the date as a random effect (see model choice in
generalized model taking into account the proportion of nauplii consumed reported to the
total number of tentacles, the tumorous status was included as a fixed factor (see model
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The proportion of tumorous to non-tumorous hydras colonized by ciliates was compared
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to the number expected under a no preference hypothesis (50%, binomial test). The
verification has been done that there is no effect of the day of measurement using a Fisher
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exact test. We used a Poisson glmm to compare the number of ciliates in each group across
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time (in days) with the group as a fixed effect and the individual as a random effect (see
supplementary material).
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The proportion of tumorous hydra captured by the predator was compared to the expected
number under a random choice, using a binomial test. The verification has been done that
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there was no effect of the measurement day, using a Fisher exact test. We excluded from the
analysis fish that attacked none of the hydras. The delay between the first and the second prey
consumption was compared using generalized model with a Poisson law (see supplementary
material). All analyses were performed using Rstudio (version 1.3)(RStudio Team, 2021) and
R (version 4.1.0)(R Core Team, 2017). The list of packages used and the explained code are
Results
(a) Hydra predation ability
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The number of tentacles was significantly higher in the tumorous group (9.35 ± 1.14 for
tumorous, 5.71 ± 0.75 for non-tumorous; mean ± standard deviation, W-value = 13, p< 2.2e-
16). Since the chief function of the tentacles is capturing prey, we hypothesized that the
tumorous polyps might have a different predation capacity compared to the healthy polyps.
We tested this hypothesis in two feeding experiments - using ad libitum and restricted amount
of prey. In both feeding conditions, tumorous hydras captured significantly higher number of
nauplii than non-tumorous ones (ad libitum: Fig3a., Incidence ratio rates (IRR) = 1.31 ± 0.07;
Z= 5.245; df= 81, p<0.001); restricted feeding conditions: Fig3b., Odds ratio (OR) = 1.54 ±
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0.22; Z=3.091; df=80; p=0.002).
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We also evaluated the importance of tentacles’ number on the feeding performance by
reporting the predation abilities of each hydra to their number of tentacles. In ad libitum
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condition, tumoral hydras captured significantly less prey than non-tumoral ones reported to
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their respective number of tentacles (Fig3c., Estimate = -0.17 ± 0.08; df= 81, p=0.025). In the
restricted feeding condition, there was no significant difference (Fig3d., IRR=0.89 ± 0.15;
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When given the choice between a tumorous and a non-tumorous hydra, the ciliates colonized
preferentially the tumorous one (Fig4a., 81% ± 13%, p = 6.877e-05, alpha=0.01, binomial
exact test, n=42), without any effect of the day of measurement (p = 0.1081, alpha=0.01,
Fisher exact test, n=42). Of the 36 individuals inoculated with ciliates in the second
term observation of the colonization dynamics uncovered that the number of ciliates on hydra
increased faster over time and then reached higher densities on tumorous hydra than on the
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Siamese fish ate hydras in 37 out of 51 trials and the first hydra consumed was significantly
more often the tumorous one (Fig5a., i.e. 73% of cases, 27 times over 37 trials, p = 0.008,
binomial exact test). Nine fish preyed only on the tumorous hydra and none of them preyed
only the non-tumorous one (Fig5b.). There was no influence of the measurement day (p=
0.650, Fisher exact test, n=37). When the first hydra consumed was a non-tumorous one, the
time needed to observe the second predation event was significantly shorter than when the
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first hydra captured was the tumorous one (Fig5c., 715 ± 738 seconds vs. 55 ± 98.3 seconds;
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IRR=0.08 ± 0.04, p<0.001, df=26).
Discussion
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Ecological implications of oncogenic processes, while theoretically important in ecosystem
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functioning (Vittecoq et al., 2013), have never been studied experimentally until now. This
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study provides empirical evidence for the first time for the hypothesis that the phenotypic
and/or quantitatively, various types of biotic interactions within the ecosystem. We found that
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(i) the predation ability of tumorous hydras was increased compared to non-tumorous ones,
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which is explained by their increased number of tentacles, (ii) the tumoral phenotype is more
often colonized by a ciliate and with a faster colonization and (iii) tumorous hydras were
preferentially eaten by predatory fish. Although tumorous and non-tumorous hydras used in
our experiments differ in the presence of tumors and their phenotypic consequences, as well
as their microbiota (Rathje et al., 2020), they originated from the same ancestral polyp and
are genetically identical (clonal). Therefore, the alterations of biotic interactions reported here
are most likely caused by the phenotypic alterations induced by the tumors (i.e. non-tumorous
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The significantly higher ability of tumorous hydras to capture prey may first appear
counterintuitive given that neoplasms are usually associated with reduced host performances
(Arnal et al., 2015; Perret et al., 2020). The most parsimonious explanation here is that this
compared to non-tumorous ones (see also (Domazet-Lošo et al., 2014)). Precise causes
behind this novel phenotype are unclear and could have different origins since it could benefit
the host (e.g. compensatory mechanisms, see for instance Thomas et al., 2020)), the tumor
(e.g. host manipulation, see for instance Tissot et al., 2016) or be beneficial for both the host
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and the tumor (see for instance (Ewald, 1980)). Deciphering the molecular cross-talk between
the host and the tumor during the window when additional tentacles are growing could help
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to clarify this point (see for instance Heil, 2016). Further experiments estimating fitness
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benefits for the host and the tumor would be required to distinguish between these
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possibilities. Also, given that microbiota is altered in the tumorous polyps, and that the
microbiota alters the behavior of hydras (Murillo-Rincon et al., 2017), we cannot exclude that
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a modified neuronal activity and feeding behavior might be responsible for the more efficient
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predation of tumorous polyps. We particularly emphasize here that the higher predation
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capacity of tumorous hydras was observed in both feeding conditions, ad libitum and
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restricted feeding, whereby the limited food supply is likely the more natural condition for
hydra (Deserti et al., 2017). In any case, these results suggest that the level of resources could
manner. Indeed, while it is intuitively logic that tumorous individuals, already weakened by
the presence of tumor, would suffer more from a lack or from scarcity of prey than non-
tumorous ones, our results conversely suggest that their persistence could be facilitated in
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The higher preference of ciliates for tumor-bearing individuals, as well as their faster
replication rate on them compared to non-tumorous ones, may indicate that the formers offer
a larger living surface and/or more resources for epibiont organisms. Precise mechanisms
operating here remain however to be determined, at least four hypotheses could be tested. (i)
Ciliates prefer chemical cues specifically produced by tumor-bearing individuals, (ii) tumor-
ciliates because of their bigger size, their higher number of tentacles and/or a different
behavior associated with tumors, (iii) tumorous hydras might lack some defenses against
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ciliates and are then more vulnerable to the opportunistic colonization, (iv) tumorous hydras
might harbor more bacteria and/or release more feeding byproducts useful for ciliates due to
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their altered feeding behavior. The commensal role itself of this ciliate remains however to be
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explored (Coleman, 1966; Foissner, 1987; Kazubski, 1991; Ribi et al., 1985; Warren and
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Robson, 1998). For instance, the presence of a high number of ciliates seems to be associated
with an increased budding rate in H. oligactis, but the causes of this phenomenon are not well
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understood (Coleman, 1966). Thus, we cannot exclude that the commensal nature of the
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interaction between this ciliate and the hydra is not stable and varies with different
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parameters including the density on hydra. Further explorations of the impact of ciliates’
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density on the fitness (e.g. budding rate, survival) of tumorous and non-tumorous hydras
could help us to understand the costs and/or the benefits of these commensals for tumorous
The higher predation risk of tumor-bearing hydra could be ascribed to both visual and
chemical cues, since the fish species used here is able to exploit both, at least in sexual
selection and competition contexts (Forsatkar et al., 2017; Ingersoll et al., 1976; Romano et
al., 2017). However, given that this fish is mostly a visually hunting predator (Bando, 1991;
da Silva Souza et al., 2020) and from our observations during the experiments, we favor the
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first hypothesis, i.e. that they detect and capture preferentially bigger (tumorous) preys. The
respective role of the abnormal neoplastic mass and/or of the higher number of tentacles in
enhancing the hydra detectability could be tested in the future by removing some tentacles
from tumor-bearing individuals, and/or by grafting the upper part of tumorous hydras on non-
tumorous ones and vice versa (see for instance Kuznetsov et al., 2002). In addition, further
experiments should also focus on the possible role of the behavioral differences between
tumorous and non-tumorous hydras. For instance, tumorous polyps could not be able to
contract spontaneously and their induced contractility can be compromised. Therefore, they
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potentially could not retract themselves in case of attack and/or would have a lower ability to
detect predators (hydras are light sensitive and can detect shadows from the fish (Musio et al.,
2001).
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Beyond the experimental evidence that tumors alter biotic interactions between hydras
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and several other species, an interesting direction to explore concerns the ecological
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notably manipulative ones, have for instance shown that many ecological consequences may
result from the novel phenotype displayed by parasitized hosts (Lafferty et al., 2008; Lefèvre
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et al., 2009). This topic has been rarely addressed in the context of cancerous pathologies
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(Hamede et al., 2020; Roche et al., 2017; Vittecoq et al., 2013). Because tumorous hydras,
compared to healthy ones, are both more efficient predators and more vulnerable prey,
stronger trophic cascades could be for instance expected in the presence of tumorous hydras
in the ecosystem, a process that is also likely to speed up energy flow and nutrient cycles as
observed for other parasitic species (Mischler et al., 2016; Shurin and Seabloom, 2005; Wood
et al., 2007). Similarly, by boosting the population dynamics of their commensal ciliates,
tumorous hydras probably enhance the probability of other hydra species to be colonized by
this generalist ciliate. Further studies manipulating the prevalence of tumorous hydras in the
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field and/or in mesocosms are necessary to explore and quantify properly these ecological
consequences.
composed of two interacting organism with, one containing tumoral cells considered as a
third living entity impacting the interaction. It allows assessing the impact of tumor burden
onto multiple biotic interactions within an ecosystem. Moreover, it can provide new insights
for a mechanistic understanding of these dynamics. The simplicity of the system and its
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accessibility offers unlimited perspective for ecological, behavioral or even biochemical
studies.
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Beyond the present demonstration that the tumorigenesis has the potential to alter
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biotic interactions, further experiments manipulating in microcosms the frequency of tumor-
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possible cascade effects on the whole aquatic community. As suggested above, these
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experiments should also test if the influence of tumors in biotic interactions depends on the
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tumor may appear advantageous on particular aspects, all other things being equal. The
evolutionary significance of this principle and its impact onto the species’ life history may be
functioning. This research direction is likely to become crucial in the future given that
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Acknowledgements:
We are grateful to Lena Guimard for her pictures. We want to thanks to Karima Kissa (LIPH,
University of Montpellier) for the provision of their premises to fish experiments. This work
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supported by the doctoral fellowship of the University of Montpellier. JT was supported by
All experimental animals were treated in accordance with French and European Union
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regulations (directive 2010/63/UE). All the behavioral observations were included in the
project authorization APAFIS: 29961. No fish were injured or killed during the experiments.
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REFERENCES
u
Aktipis, C.A., Boddy, A.M., Jansen, G., Hibner, U., Hochberg, M.E., Maley, C.C.,
Jo
Wilkinson, G.S., 2015. Cancer across the tree of life: cooperation and cheating in
multicellularity. Philos. Trans. R. Soc. B Biol. Sci. 370, 20140219.
https://doi.org/10.1098/rstb.2014.0219
Aktipis, C.A., Nesse, R.M., 2013. Evolutionary foundations for cancer biology. Evol. Appl.
https://doi.org/10.1111/eva.12034
Albuquerque, T.A.F., Val, L.D. do, Doherty, A., Magalhães, J.P. de, 2018. From humans to
hydra: patterns of cancer across the tree of life. Biol. Rev. 93, 1715–1734.
https://doi.org/10.1111/brv.12415
Alcock, J., Maley, C.C., Aktipis, C.A., 2014. Is eating behavior manipulated by the
gastrointestinal microbiota? Evolutionary pressures and potential mechanisms.
16
Journal Pre-proof
BioEssays. https://doi.org/10.1002/bies.201400071
Argilés, J.M., Busquets, S., Stemmler, B., López-Soriano, F.J., 2014. Cancer cachexia:
Understanding the molecular basis. Nat. Rev. Cancer. https://doi.org/10.1038/nrc3829
Arnal, A., Jacqueline, C., Ujvari, B., Leger, L., Moreno, C., Faugere, D., Tasiemski, A.,
Boidin-Wichlacz, C., Misse, D., Renaud, F., Montagne, J., Casali, A., Roche, B., Mery,
F., Thomas, F., 2017. Cancer brings forward oviposition in the fly Drosophila}
melanogaster. Ecol. Evol. 7, 272–276. https://doi.org/10.1002/ece3.2571
Arnal, A., Ujvari, B., Crespi, B., Gatenby, R.A., Tissot, T., Vittecoq, M., Ewald, P.W.,
Casali, A., Ducasse, H., Jacqueline, C., Missé, D., Renaud, F., Roche, B., Thomas, F.,
f
oo
2015. Evolutionary perspective of cancer: myth, metaphors, and reality. Evol Appl 8,
541–544. https://doi.org/10.1111/eva.12265
pr
Bando, T., 1991. Visual perception of texture in aggressive behavior of Betta splendens. J.
Comp. Physiol. A. https://doi.org/10.1007/BF00198172
e-
Barr, R.D., Ries, L.A.G., Lewis, D.R., Harlan, L.C., Keegan, T.H.M., Pollock, B.H., Bleyer,
Pr
W.A., 2016. Incidence and incidence trends of the most frequent cancers in adolescent
and young adult Americans, including “nonmalignant/noninvasive” tumors. Cancer 122,
al
1000–1008. https://doi.org/10.1002/cncr.29867
rn
Baumga, D., Constance, L., 2005. The impact of predation by burbot ( Lota lota L .) on the
macroinvertebrate community in the littoral zone of a large lake. Aquat. Ecol. 79–92.
u
Buszewski, B., Rudnicka, J., Ligor, T., Walczak, M., Jezierski, T., Amann, A., 2012.
Jo
Analytical and unconventional methods of cancer detection using odor. TrAC - Trends
Anal. Chem. https://doi.org/10.1016/j.trac.2012.03.019
Chalmandrier, L., Albouy, C., Pellissier, L., 2017. Species pool distributions along functional
trade-offs shape plant productivity–diversity relationships. Sci. Rep. 7.
https://doi.org/10.1038/s41598-017-15334-4
Coleman, D.C., 1966. The Laboratory Population Ecology of Kerona Pediculus (O.F.M.)
Epizoic on Hydra SPP. Ecology. https://doi.org/10.2307/1934258
Cuker, B.E., Mozley, S.C., 1981a. Summer population fluctuations, feeding, and growth of
Hydra in an arctic lake. Limnol. Oceanogr. https://doi.org/10.4319/lo.1981.26.4.0697
17
Journal Pre-proof
Cuker, B.E., Mozley, S.C., 1981b. Summer population fluctuations, feeding, and growth of
Hydra in an arctic lake. Limnol. Oceanogr. 26, 697–708.
https://doi.org/10.4319/lo.1981.26.4.0697
da Silva Souza, J.G., Libeck, L.T., do Carmo Rodrigues Virote, B., Egger, R.C., de Sá,
G.C.R., Machado, G.J., Murgas, L.D.S., 2020. A method to analyze the relationship
between locomotor activity and feeding behaviour in larvae of Betta splendens. Aquac.
Int. https://doi.org/10.1007/s10499-020-00516-1
Dawson, E.H., Bailly, T.P.M., Dos Santos, J., Moreno, C., Devilliers, M., Maroni, B., Sueur,
C., Casali, A., Ujvari, B., Thomas, F., Montagne, J., Mery, F., 2018. Social environment
f
mediates cancer progression in Drosophila. Nat. Commun.
oo
https://doi.org/10.1038/s41467-018-05737-w
pr
Deserti, M.I., Esquius, K.S., Escalante, A.H., Acuña, F.H., 2017. Trophic ecology and diet of
Hydra vulgaris (Cnidaria; Hydrozoa). Anim. Biol. https://doi.org/10.1163/15707563-
e-
00002537
Pr
Domazet-Lošo, T., Klimovich, A., Anokhin, B., Anton-Erxleben, F., Hamm, M.J., Lange, C.,
Bosch, T.C.G., 2014. Naturally occurring tumours in the basal metazoan {Hydra}. Nat
Commun 5, 4222. https://doi.org/10.1038/ncomms5222
al
Elliott, J.K., Elliott, J.M., Leggett, W.C., 1997. Predation by Hydra on larval fish: Field and
rn
Ewald, P.W., 1980. Evolutionary biology and the treatment of signs and symptoms of
infectious disease. J. Theor. Biol. https://doi.org/10.1016/0022-5193(80)90073-9
Foissner, W., 1987. Miscellanea Nomenclatorica Ciliatea (Protozoa: Ciliophora). Arch. fur
Protistenkd. https://doi.org/10.1016/S0003-9365(87)80054-4
Forsatkar, M.N., Nematollahi, M.A., Brown, C., 2017. Male Siamese fighting fish use gill
flaring as the first display towards territorial intruders. J. Ethol.
https://doi.org/10.1007/s10164-016-0489-1
Giraudeau, M., Sepp, T., Ujvari, B., Ewald, P.W., Thomas, F., 2018. Human activities might
influence oncogenic processes in wild animal populations. Nat. Ecol. Evol.
https://doi.org/10.1038/s41559-018-0558-7
18
Journal Pre-proof
Hamede, R., Owen, R., Siddle, H., Peck, S., Jones, M., Dujon, A.M., Giraudeau, M., Roche,
B., Ujvari, B., Thomas, F., 2020. The ecology and evolution of wildlife cancers:
Applications for management and conservation. Evol. Appl. 13, 1719–1732.
https://doi.org/10.1111/EVA.12948
Hamilton, D.G., Jones, M.E., Cameron, E.Z., Kerlin, D.H., McCallum, H., Storfer, A.,
Hohenlohe, P.A., Hamede, R.K., 2020. Infectious disease and sickness behaviour:
tumour progression affects interaction patterns and social network structure in wild
Tasmanian devils. Proc. R. Soc. B Biol. Sci. 287, 20202454.
https://doi.org/10.1098/rspb.2020.2454
f
Heil, M., 2016. Host manipulation by parasites: Cases, patterns, and remaining doubts. Front.
oo
Ecol. Evol. https://doi.org/10.3389/fevo.2016.00080
pr
Hughes, D.P., Brodeur, J., Thomas, F. (Eds.), 2012. Host Manipulation by Parasites, Host
Manipulation by Parasites. Oxford University Press.
e-
https://doi.org/10.1093/acprof:oso/9780199642236.001.0001
Pr
Hughes, D.P., Libersat, F., 2019. Parasite manipulation of host behavior. Curr. Biol.
https://doi.org/10.1016/j.cub.2018.12.001
al
Ingersoll, D.W., Bronstein, P.M., Bonventre, J., 1976. Chemical modulation of agonistic
display in Betta splendens. J. Comp. Physiol. Psychol. https://doi.org/10.1037/h0077195
rn
Jones, K., Ariel, E., Burgess, G., Read, M., 2016. A review of fibropapillomatosis in Green
u
Kazubski, S., 1991. Morphological variation of the ciliate Trichodina pediculus Ehrenberg,
1838. I : Parasitizing on hydras. Acta Protozool.
https://repository.naturalis.nl/pub/317635
Kishida, O., Trussell, G.C., Mougi, A., Nishimura, K., 2010. Evolutionary ecology of
inducible morphological plasticity in predator-prey interaction: Toward the practical
links with population ecology. Popul. Ecol. https://doi.org/10.1007/s10144-009-0182-0
Kuznetsov, S.G., Anton-Erxleben, F., Bosch, T.C.G., 2002. Epithelial interactions in Hydra:
Apoptosis in interspecies grafts is induced by detachment from the extracellular matrix.
J. Exp. Biol.
Lafferty, K.D., Allesina, S., Arim, M., Briggs, C.J., Leo, G. De, Dobson, A.P., Dunne, J.A.,
19
Journal Pre-proof
Johnson, P.T.J., Kuris, A.M., Marcogliese, D.J., Martinez, N.D., Memmott, J., Marquet,
P.A., McLaughlin, J.P., Mordecai, E.A., Pascual, M., Poulin, R., Thieltges, D.W., 2008.
Parasites in food webs: the ultimate missing links. Ecol. Lett. 11, 533–546.
https://doi.org/10.1111/J.1461-0248.2008.01174.X
Lefèvre, T., Lebarbenchon, C., Gauthier-Clerc, M., Missé, D., Poulin, R., Thomas, F., 2009.
The ecological significance of manipulative parasites. Trends Ecol. Evol. 24, 41–48.
https://doi.org/10.1016/J.TREE.2008.08.007
Lenhoff, H.M., Brown, R.D., 1970. Mass culture of hydra: an improved method and its
application to other aquatic invertebrates. Lab Anim 4, 139–154.
f
https://doi.org/10.1258/002367770781036463
oo
Loreau, M., 2010. Linking biodiversity and ecosystems: Towards a unifying ecological
pr
theory. Philos. Trans. R. Soc. B Biol. Sci. https://doi.org/10.1098/rstb.2009.0155
Madsen, T., Arnal, A., Vittecoq, M., Bernex, F., Abadie, J., Labrut, S., Garcia, D., Faugère,
e-
D., Lemberger, K., Beckmann, C., Roche, B., Thomas, F., Ujvari, B., 2017. Cancer
Pr
Prevalence and Etiology in Wild and Captive Animals, Ecology and Evolution of
Cancer. Elsevier Inc. https://doi.org/10.1016/B978-0-12-804310-3.00002-8
al
Massol, F., Gravel, D., Mouquet, N., Cadotte, M.W., Fukami, T., Leibold, M.A., 2011.
Linking community and ecosystem dynamics through spatial ecology. Ecol. Lett.
rn
https://doi.org/10.1111/j.1461-0248.2011.01588.x
u
Mischler, J., Johnson, P.T.J., McKenzie, V.J., Townsend, A.R., 2016. Parasite infection alters
Jo
Moore, J., 2013. An overview of parasite-induced behavioral alterations - and some lessons
from bats. J. Exp. Biol. https://doi.org/10.1242/jeb.074088
Murillo-Rincon, A.P., Klimovich, A., Pemöller, E., Taubenheim, J., Mortzfeld, B., Augustin,
R., Bosch, T.C.G., 2017. Spontaneous body contractions are modulated by the
microbiome of Hydra. Sci. Rep. https://doi.org/10.1038/s41598-017-16191-x
Musio, C., Santillo, S., Taddei-Ferretti, C., Robles, L.J., Vismara, R., Barsanti, L., Gualtieri,
20
Journal Pre-proof
P., 2001. First identification and localization of a visual pigment in Hydra (Cnidaria,
Hydrozoa). J. Comp. Physiol. - A Sensory, Neural, Behav. Physiol.
https://doi.org/10.1007/s003590100180
Pascal, L., Grémare, A., de Montaudouin, X., Deflandre, B., Romero-Ramirez, A., Maire, O.,
2020. Parasitism in ecosystem engineer species: A key factor controlling marine
ecosystem functioning. J. Anim. Ecol. https://doi.org/10.1111/1365-2656.13236
Perret, C., Gidoin, C., Ujvari, B., Thomas, F., Roche, B., 2020. Predation shapes the impact
of cancer on population dynamics and the evolution of cancer resistance. Evol. Appl. 13,
1733–1744. https://doi.org/10.1111/eva.12951
f
oo
Petchey, O.L., Gaston, K.J., 2002. Functional diversity (FD), species richness and community
composition. Ecol. Lett. https://doi.org/10.1046/j.1461-0248.2002.00339.x
pr
R Core Team, 2017. R: A Language and Environment for Statistical Computing. Vienna: R
Core Team.http://r.meteo.uni.wroc.pl/web/packages/dplR/vignettes/intro-dplR.pdf
e-
Raffard, A., Santoul, F., Cucherousset, J., Blanchet, S., 2019. The community and ecosystem
Pr
Rathje, K., Mortzfeld, B., Hoeppner, M.P., Taubenheim, J., Bosch, T.C.G., Klimovich, A.,
rn
2020. Dynamic interactions within the host-associated microbiota cause tumor formation
in the basal metazoan Hydra. PLoS Pathog.
u
https://doi.org/10.1371/journal.ppat.1008375
Jo
Reed, T.E., Robin, S.W., Schindler, D.E., Hard, J.J., Kinnison, M.T., 2010. Phenotypic
plasticity and population viability: The importance of environmental predictability. Proc.
R. Soc. B Biol. Sci. https://doi.org/10.1098/rspb.2010.0771
Ribi, G., Tardent, R., Tardent, P., Scascighini, C., 1985. Dynamics of hydra populations in
Lake Zürich, Switzerland, and Lake Maggiore, Italy. Swiss J. Hydrol.
https://doi.org/10.1007/BF02538183
Rivera-De la Parra, L., Sarma, S.S.S., Nandini, S., 2016. Effects of predation by Hydra
(Cnidaria) on cladocerans (Crustacea: Cladocera). J. Limnol.
https://doi.org/10.4081/jlimnol.2016.1368
Roche, B., Mller, A.P., DeGregori, J., Thomas, F., 2017. Cancer in Animals: Reciprocal
21
Journal Pre-proof
Romano, D., Benelli, G., Donati, E., Remorini, D., Canale, A., Stefanini, C., 2017. Multiple
cues produced by a robotic fish modulate aggressive behaviour in Siamese fighting
fishes. Sci. Rep. 7, 1–11. https://doi.org/10.1038/s41598-017-04840-0
Sharpe, S., 2020. Combating the Hydra; An Aquarium Pest. Spruce Pets.
https://doi.org/www.thesprucepets.com/combat-aquarium-pest-hydra-1381228
f
oo
Shurin, J.B., Seabloom, E.W., 2005. The strength of trophic cascades across ecosystems:
predictions from allometry and energetics. J. Anim. Ecol. 74, 1029–1038.
pr
https://doi.org/10.1111/J.1365-2656.2005.00999.X
e-
Stiven, A.E., 1965. The relationship between size, budding rate, and growth efficiency in
three species of hydra. Res. Popul. Ecol. (Kyoto). 7, 1–15.
Pr
https://doi.org/10.1007/BF02518810
Thomas, F., Brodeur, J., Maure, F., Franceschi, N., Blanchet, S., Rigaud, T., 2011.
al
https://doi.org/10.1016/j.meegid.2010.12.013
Thomas, F., Giraudeau, M., Dheilly, N.M., Gouzerh, F., Boutry, J., Beckmann, C., Biro, P.A.,
u
Hamede, R., Abadie, J., Labrut, S., Bieuville, M., Misse, D., Bramwell, G., Schultz, A.,
Jo
Le Loc’h, G., Vincze, O., Roche, B., Renaud, F., Russell, T., Ujvari, B., 2020. Rare and
unique adaptations to cancer in domesticated species: An untapped resource? Evol.
Appl. https://doi.org/10.1111/eva.12920
Thomas, F., Jacqueline, C., Tissot, T., Henard, M., Blanchet, S., Loot, G., Dawson, E., Mery,
F., Renaud, F., Montagne, J., Beckmann, C., Biro, P.A., Hamede, R., Ujvari, B., 2017.
The importance of cancer cells for animal evolutionary ecology. Nat. Ecol. Evol. 1,
1592–1595. https://doi.org/10.1038/s41559-017-0343-z
Thomas, F., Renaud, F., de Meeus, T., Poulin, R., 1998. Manipulation of host behaviour by
parasites: Ecosystem engineering in the intertidal zone? Proc. R. Soc. B Biol. Sci. 265,
1091–1096. https://doi.org/10.1098/rspb.1998.0403
22
Journal Pre-proof
Thompson, R.M., Brose, U., Dunne, J.A., Hall, R.O., Hladyz, S., Kitching, R.L., Martinez,
N.D., Rantala, H., Romanuk, T.N., Stouffer, D.B., Tylianakis, J.M., 2012. Food webs:
Reconciling the structure and function of biodiversity. Trends Ecol. Evol.
https://doi.org/10.1016/j.tree.2012.08.005
Tissot, T., Arnal, A., Jacqueline, C., Poulin, R., Lefèvre, T., Mery, F., Renaud, F., Roche, B.,
Massol, F., Salzet, M., Ewald, P., Tasiemski, A., Ujvari, B., Thomas, F., 2016. Host
manipulation by cancer cells: {Expectations}, facts, and therapeutic implications.
BioEssays 38, 276–285. https://doi.org/10.1002/bies.201500163
Vittecoq, M., Roche, B., Daoust, S.P., Ducasse, H., Missé, D., Abadie, J., Labrut, S., Renaud,
f
F., Gauthier-Clerc, M., Thomas, F., 2013. Cancer: A missing link in ecosystem
oo
functioning? Trends Ecol. Evol. https://doi.org/10.1016/j.tree.2013.07.005
pr
Walters, A.W., Hughes, R.C., Call, T.B., Walker, C.J., Wilcox, H., Petersen, S.C., Rudman,
S.M., Newell, P.D., Douglas, A.E., Schmidt, P.S., Chaston, J.M., 2020. The microbiota
e-
influences the Drosophila melanogaster life history strategy. Mol. Ecol.
https://doi.org/10.1111/mec.15344
Pr
Warren, A., Robson, E. a, 1998. The identity and occurrence of Kerona pediculus (
Ciliophora : Hypotrichida ), a well-known epizoite of Hydra vulgaris ( Cnidaria :
al
Wong, A.C.N., Holmes, A., Ponton, F., Lihoreau, M., Wilson, K., Raubenheimer, D.,
Simpson, S.J., 2015. Behavioral microbiomics: A multi-dimensional approach to
u
https://doi.org/10.3389/fmicb.2015.01359
Wood, C.L., Byers, J.E., Cottingham, K.L., Altman, I., Donahue, M.J., H Blakeslee, A.M.,
2007. Parasites alter community structure.
Wright, J.P., Ames, G.M., Mitchell, R.M., 2016. The more things change, the more they stay
the same? When is trait variability important for stability of ecosystem function in a
changing environment. Philos. Trans. R. Soc. B Biol. Sci.
https://doi.org/10.1098/rstb.2015.0272
Zuur, A.F., Ieno, E.N., Walker, N.J., Saveliev, A.A., York, G.M.S.N., 2013. Mixed Effects
Models and Extensions in Ecology with R 1. https://doi.org/ 10.1007/978-0-387-87458-
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Legend of figures
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individual from a lineage established after a field sampled individual at Tiszadorogma in
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Hungary, (47°67’12”N, 20°86’41”E) that is prone to develop tumors spontaneously in routine
cultures.
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Figure 2: Phenotypic differences of non-tumorous and tumorous hydras
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The non-tumorous hydra is on the left and the tumorous one on the right. We can notice an
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(a.) The number of prey captured per hydra in ad libitum, (b.) the proportion of prey captured
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per hydra in restricted feeding, (c.) the number of prey captured per tentacle in ad libitum
and, (d.) the proportion of prey captured per tentacle in restricted feeding (b.) were counted
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during ten minutes. Boxplots represent the first, median and third quartiles on the 95% extent.
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A point represents each measurement with the non-tumorous group in green (on the left) and
the tumorous in red (on the right). (e.) Pictures of tumorous and non-tumorous hydras
capturing prey.
(a.) Five hours after the introduction of a ciliate into an experimental tank containing both
hydra type, we found 81% of the ciliates attaches to the tumoral host instead of the non-
tumoral one. (b.) After the introductions of one ciliate into the well of a tumoral or a non-
tumoral individual, we counted daily the number of ciliates per hydra. Each point represents
the number of ciliates measured on a given day; the size of the point depends on the number
of hydras recorded at this measurement. The black line represents the average value predicted
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by the model and the colored areas the confidence interval at 95% predicted by the model. In
green, on the left, the non-tumorous group on the left and in red, on the right, the tumorous
group on the right in red. (c.) Pictures of eleven ciliates attached to a hydra at x20 and
zoomed picture of one ciliate on a hydra.
Figure 5: Tumorous hydras have a higher predation risk by fish than non-tumorous
ones
(a.) The histogram represents the number of fish that consumed first the tumoral (in red), the
non-tumoral (in green) or none of the hydra during the test (in grey). Among the 37 trials
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which fish consume a first hydra, the pie plots represent the proportion of fish that consumed
the second one. The color of the section corresponds the type of the remaining hydra; in grey
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if the second hydra consumed is non-tumoral, in red if the second hydra consumed is tumoral
and in grey if the fish did not consume the second hydra. (c.) A point represents the time
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required to the fish to consume the second hydra after the first one, on the left when the
remaining prey is non-tumorous (in green), on the right when it is a tumorous (in red). (d.)
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Pictures of two hydras, a tumoral and a non-tumoral in a plastic well plate. The fish
consumed the tumoral, and then, only the non-tumoral remain visible on the last picture.
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These illustrative pictures were taken during a session separate from the experiments.
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Figures
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Figure 1. Pictures of different individuals harboring spontaneous tumors in strains
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established from fields sampling.
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Figure 5. Tumorous hydras have a higher predation risk by fish than non-tumorous
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Declaration of interests
☒ The authors declare that they have no known competing financial interests or personal
relationships that could have appeared to influence the work reported in this paper.
☐The authors declare the following financial interests/personal relationships which may be
considered as potential competing interests:
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Graphical abstract
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HIGHLIGHTS:
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