Ecological Indicators 107 (2019) 105672

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Ecological Indicators
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Epibiotic barnacles of sea turtles as indicators of habitat use and fishery T

interactions: An analysis of juvenile loggerhead sea turtles, Caretta caretta, in
the western Mediterranean
S. Ten, L. Pascual, M.I. Pérez-Gabaldón, J. Tomás, F. Domènech, F.J. Aznar
⁎

Marine Zoology Unit, Cavanilles Institute of Biodiversity and Evolutionary Biology, University of Valencia, 46980, Paterna, Valencia, Spain

ARTICLE INFO ABSTRACT

Keywords: Sea turtles exploit a variety of habitats during their lifetime and are thus exposed to a number of anthropogenic
Sea turtle threats, including interaction with fisheries. Mitigating this impact requires determining patterns of habitat use
Caretta caretta by turtles, which largely relies on data from marked individuals. We investigated the use of epibiotic barnacles as
Habitat a cost-effective method to ascertain habitat use by sea turtles, choosing the juvenile loggerhead sea turtles
Fisheries
(Caretta caretta) in the western Mediterranean as a model. A total of 200 turtles collected in the central Spanish
Barnacle
Epibiont
Mediterranean between 1990 and 2017 were examined for epibionts: 25% were accidentally captured by pelagic
Indicator longliners, 19% by neritic gears, and 56% were stranded by unknown causes. Gut contents of 80 deceased turtles
were also examined to infer the habitat(s) exploited by the turtles 1–2 weeks prior to death. We collected 16,652
barnacles of 13 species. Barnacle assemblages of turtles captured by longliners, or having oceanic prey only,
were dominated by Lepas hillii and Conchoderma virgatum, whereas the turtles captured by neritic fisheries, or
having neritic prey, had significantly higher loads of Chelonibia testudinaria. A canonical discriminant analysis
based on barnacle abundance correctly assigned 80.5% of caught turtles to the responsible fishery, and predicted
a nearly identical assignation to fishery of stranded turtles. The estimated age distribution for C. virgatum was
unimodal, and many individuals were inferred to have attached postmortem, thus tracing the habitat where
turtles died. The age distribution for L. hillii was bimodal, and the old individuals informed on stays of turtles in
oceanic waters of at least few weeks prior to death. Finally, C. testudinaria settled on live turtles and were
comparatively older, marking the stay of turtles in neritic waters several months before death. In summary, data
on presence and estimated age of these 3 barnacle species not only shed much light on habitat where turtles died,
but also helped to trace movements and residence times in each habitat. We encourage researchers to include
barnacles as a complementary source of data on habitat use of sea turtles.

1. Introduction (e.g., turtle excluder devices) and improvement of fishery regulations

Sea turtles are an integral element of marine ecosystems: they play a
significant ecological role by maintaining healthy habitats (Hannan
et al., 2007; Goatley et al., 2012; Hearne et al., 2019), providing nu-
trients to other marine species (Sazima et al., 2004; Schofield et al.,
2006), controlling jellyfish populations (Gibbons and Richardson,
2008) and hosting epibiont communities (Frick and Pfaller, 2013).
Unfortunately, six of the seven sea turtle species are currently threa-
tened (IUCN, 2019). Among the numerous threats faced by sea turtles,
incidental capture by fisheries generates one of the highest population
impacts (Wallace et al., 2013). Much effort has been made to reduce sea
turtle mortality associated with fisheries through gear modifications
(Hamann et al., 2010; Rees et al., 2016). A key element to minimize
turtle bycatch is to investigate movements and patterns of habitat use
by turtles so that fisheries exploit areas/habitats with lower probability
of turtle encounters (Hamann et al., 2010).
The western Mediterranean represents an important feeding area for
juveniles of loggerhead sea turtle (Caretta caretta) that come from one
Mediterranean and two Atlantic Regional Management Units (Monzón-
Argüello et al., 2010; Wallace et al., 2010; Carreras et al., 2011; Clusa
et al., 2014). In the western Mediterranean, loggerhead turtles have
been severely impacted by fisheries for several decades (Aguilar et al.,
1995; Carreras et al., 2004; Camiñas et al., 2006; Báez et al., 2007a,b).
Between 1995 and 2004, over 3400 loggerhead turtles were

⁎
Corresponding author.
E-mail addresses: sotengon@alumni.uv.es (S. Ten), maipega@alumni.uv.es (M.I. Pérez-Gabaldón), jesus.tomas@uv.es (J. Tomás),
francesc.domenech@uv.es (F. Domènech), francisco.Aznar@uv.es (F.J. Aznar).

https://doi.org/10.1016/j.ecolind.2019.105672
Received 3 July 2019; Received in revised form 21 August 2019; Accepted 23 August 2019
Available online 06 September 2019
1470-160X/ © 2019 Elsevier Ltd. All rights reserved.
S. Ten, et al.
incidentally captured by drifting longlines in oceanic waters (Camiñas
et al., 2006). Although this impact has recently been reduced (Camiñas
et al., 2006; Tomás et al., 2008; Báez et al., 2019), the mortality as-
sociated to neritic fisheries (mainly bottom trawling, trammel nets, and
bottom longline) is still substantial (> 650 turtles per year in the
Spanish Mediterranean; Carreras et al., 2004; Álvarez de Quevedo et al.,
2006; Domènech et al., 2015a).
Traditionally, it has been assumed that juvenile loggerhead turtles
undergo a deterministic ontogenetic habitat shift, with early and late
juveniles exploiting oceanic and neritic waters, respectively (Bolten,
2003). However, at least in some populations, this shift appears to vary
more idiosyncratically among individuals depending on food avail-
ability and oceanographic features (Casale et al., 2008, 2012a). In the
western Mediterranean, many juvenile loggerhead turtles of different
ages undergo repeated short-term shifts between oceanic and neritic
waters (e.g., Eckert et al., 2008; Cardona et al., 2009; Abalo-Morla
et al., 2018). This element of unpredictability makes it difficult to im-
plement effective bycatch mitigation measures that target specific size/
age population segments (see, e.g., Báez et al., 2013), and stresses the
need for gathering more data on habitat use by juvenile loggerheads.
Data on habitat use and movements of loggerhead turtles in the
western Mediterranean have been obtained through several methods,
including mark-recapture techniques (Revellés et al., 2008), aerial
surveys (Gómez de Segura et al., 2006), dietary analyses (Tomás et al.,
2001; Revellés et al., 2007a), genetic markers (Clusa et al., 2016), and
satellite telemetry (Cardona et al., 2005, 2009; Revellés et al., 2007b;
Eckert et al., 2008). A priori, epibiotic barnacles could also represent a
cost-effective indicator of habitat exploitation by sea turtles (Hayashi,
2009), but their use is still circumstantial in most cases (Stamper et al.,
2005; Koch and Willan, 2007). To our knowledge, only Casale et al.
(2004, 2012b) has attempted to draw inferences on habitat use based
on a qualitative assessment of barnacle assemblages from a large
sample of loggerhead turtles in the central Mediterranean. These au-
thors suggested that four barnacle species, namely, the stalked barna-
cles (Lepadomorpha) Lepas anatifera and Conchoderma virgatum, and
two acorn barnacles (Balanomorpha), i.e., Platylepas spp., would be
more typical from turtles floating at sea or caught by pelagic longlines.
Another four balanomorphs, i.e., Stomatolepas elegans, Stephanolepas
muricata and two Chelonibia spp. would tend to be associated with
turtles caught by demersal neritic gears.
In this study we assessed the use of barnacles as ecological in-
dicators of habitat use by loggerhead turtles in the western
Mediterranean following a comprehensive approach. First, we com-
pared frequency of occurrence and abundance of epibiotic barnacles of
loggerhead turtles incidentally captured by oceanic and neritic fish-
eries, and applied the evidence obtained to infer the putative habitat of
death for turtles that stranded for unknown causes. Second, we com-
pared barnacle assemblages for samples of turtles with different di-
gestive contents, i.e., with oceanic-pelagic prey only, neritic-benthic
prey only, or both. This comparison shed light on the habitat(s) that the
turtles exploited 1–2 weeks prior to death (see Di Bello et al., 2006;
Valente et al., 2008). Finally, we used the estimated age of indicator
barnacles to trace habitat use by turtles at a finer temporal scale. Here,
we adopted a very similar methodological approach compared to that
used in studies on seaweed dispersion (Fraser et al., 2011; López et al.,
2017) or human forensics (Magni et al., 2015).
2. Methods
2.1. Study area and sample collection
Two hundred loggerhead turtles were collected between 1990 and
2017 in waters and coasts of the Valencian Community, east Spain
(between 40°101 31.5′ N, 0° 31.0′ E and 37° 50.7′ N, 1° 37.5′ W of
coastline). Turtles were found stranded (n = 112), or were incidentally
captured by pelagic longliners (n = 50) or benthic-neritic fisheries,
2
Ecological Indicators 107 (2019) 105672
including bottom trawling (n = 18) and artisanal fisheries (n = 20).
Curved carapace length notch-to-tip (CCL) ranged from 11 to 80 cm
(mean ± SD: 53.5 ± 14.2, n = 179; information on CCL was missing
from 21 individuals). Since the CCL at maturity is c. 80 cm for
Mediterranean loggerhead turtles (Casale et al., 2011), we assumed that
most turtles in our sample, if not all, were juveniles. Turtles were
classified according to standard decomposition codes sensu Santos et al.
(2018): 0, alive (n = 48); 1, freshly dead (n = 65); 2, moderately de-
composed (n = 31), 3, in advanced decomposition (n = 45). Decom-
position codes could not be determined for 11 turtles.
Epibiotic barnacles were removed during the necropsy, identified,
counted, and their body size measured with a digital caliper to the
nearest 0.01 mm. Capitulum length and maximum basal diameter
(rostro-carinal length) were used as proxies for size in lepadomorph and
balanomorph barnacles, respectively (Inatsuchi et al., 2010; Doell et al.,
2017).
In 80 turtles, the whole digestive tract was frozen for dietary ana-
lysis. After thawing, the contents from the stomach and the intestine
were separately emptied into a 0.40 mm mesh sieve and rinsed with
water. Food items were preserved dried or in 70% ethanol, and later
identified to the lowest taxon possible. For the purpose of this study, we
classified food items into pelagic-oceanic (which included remains of
gelatinous plankton, i.e., pyrosomids, salpids, doliolids and jellyfish)
and benthic-neritic (which included remains of benthic invertebrates,
mostly hermit crabs, gastropods and bivalves). Thus, each turtle was
assigned to one of three categories: (1) with pelagic-oceanic prey only;
(2) with benthic-neritic prey only, and (3) with both types of prey. A
detailed account of the prey taxa found is beyond the scope of the
present study and will be provided elsewhere (Domènech, manuscript
in prep.).
2.2. Statistical analyses
Frequency of occurrence and mean abundance per turtle were cal-
culated for each barnacle species. Confidence intervals (95%) for the
frequency of occurrence were calculated using Sterne’s exact method
(Reiczigel, 2003) and for mean abundance with accelerated and bias-
corrected bootstrap with 10,000 replicates (Rózsa et al., 2000). Pair-
wise associations between the most prevalent barnacle species were
examined with Fisher tests (for occurrence data) and Spearman corre-
lations (for abundance data), using turtles harbouring at least one
barnacle species of each pair. We also investigated whether the average
size of barnacles per turtle co-varied between barnacle species using
Spearman correlations.
A generalized lineal model (GLM) with binomial error was used to
investigate whether the probability that turtles had barnacles was in-
fluenced by CCL (fixed covariate), season (fixed factor) and provenance köken,
kaynak
of turtle (fixed factor with two groups: fishery interaction vs. stranding
of unknown cause). Turtles with missing information for CCL were
excluded from the analysis. The full model included two-way interac-
tions between predictors. Non-significant interactions were removed
from the final model to increase the sensitivity of the analysis and to
correctly interpret main effects (Engqvist, 2005).
Overall differences in barnacle assemblages between turtles cap-
tured by different fisheries (fixed factor), i.e., pelagic longliners vs.
neritic fisheries (bottom trawling and artisanal gears) were analyzed
with a permutational analysis of covariance (PERMANCOVA). Season
(fixed factor) and CCL (fixed covariate) were included as additional
predictors to control for potential confounding effects. A second PER-
MANCOVA was used to compare barnacle assemblages between turtles
with different digestive contents (fixed factor), i.e., pelagic-oceanic
prey only, benthic-neritic prey only, or both types of prey. Season (fixed
factor) and CCL (fixed covariate) were also included as additional
predictors. Non-significant interactions were removed from final ana-
lysis. Barnacle abundances were firstly fourth-root transformed and a
Bray-Curtis similarity matrix between was obtained. Pseudo-F statistics
S. Ten, et al. Ecological Indicators 107 (2019) 105672

Table 1
Frequency of occurrence (%) and abundance of epibiotic barnacles collected from a sample of 200 loggerhead sea turtles, Caretta caretta, that were found stranded or
incidentally captured in the western Mediterranean between 1990 and 2017. Barnacle species are classified according to Domènech et al. (2015b) as obligate
commensals (which attach only to living substrata) and facultative commensals (which are able to colonize a high variety of living and non-living substrata).
Barnacle species Prevalence (95% CI) Mean abundance per turtle (95% CI) Total abundance

Obligate commensals
Platylepas hexastylos 61.0 (54.3–67.9) 30.7 (18.5–54.40) 6117
Chelonibia testudinaria 8.5 (5.2–13.3) 5.24 (1.84–13.80) 1042
Chelonibia caretta 0.5 (0.0–2.9) 0.01 (0.00–0.04) 3
Stomatolepas elegans 0.5 (0.0–2.9) 0.01 (0.00–0.03) 2
Stephanolepas muricata 0.5 (0.0–2.9) 0.07 (0.00–0.21) 14

Facultative commensals
Lepas hillii 36.5 (30.1–43.7) 39.7 (23.8–68.30) 7907
Conchoderma virgatum 21.0 (15.8–27.5) 5.71 (3.37–9.75) 1131
Lepas pectinata 2.5 (1.0–5.7) 0.17 (0.03–0.48) 33
Lepas anserifera 1.5 (0.4–4.4) 0.02 (0.00–0.05) 4
Perforatus perforatus 1.5 (0.4–4.4) 0.04 (0.01–0.13) 9
Lepas anatifera 1.5 (0.4–4.4) 0.11 (0.01–0.49) 22
Amphibalanus amphitrite 1.0 (0.2–3.7) 0.05 (0.00–0.16) 10
Balanus trigonus 1.0 (0.2–3.7) 0.04 (0.00–0.18) 9

under a true null hypothesis were obtained by using 20,000 permuta-
tions (see Anderson et al., 2008). To check the validity of inferences
regarding fishery or dietary effects, tests of homogeneity of dispersion
based on distances to centroids were also carried out (Anderson et al.,
2008). The contribution of each barnacle species to the separation be-
tween fisheries or diets was examined through SIMPER decompositions
of average Bray-Curtis dissimilarities (Clarke and Gorley, 2006). Ab-
solute numerical differences of barnacle species between fisheries or
dietary categories were examined with Mann-Whitney and Kruskal-
Wallis tests, respectively. The relationship between CCL and the
abundance of each barnacle species was tested with Spearman corre-
lation tests.
A Canonical Discriminant Analysis (CDA) was also carried out to
predict the habitat of capture (oceanic vs. neritic) based on the abun-
dance of the three species that could be potential habitat indicators, i.e.,
Lepas hillii, Conchoderma virgatum and Chelonibia testudinaria (see
Results). Barnacle abundances (+1) were log10-transformed prior to
analysis. Prediction success was evaluated based on the percentage of
correct classification after jack-knife resampling. The discriminant
function obtained was also used to predict the putative habitat of
capture for stranded turtles, assuming that they had also interacted
with fisheries.
Calculations of infection parameters were made with the open ac-
cess program Quantitative Parasitology QPweb v.1.0.13 (Reiczigel et al.,
2013). PERMANCOVA and SIMPER analyses were carried out with the
statistical package PERMANOVA+ for PRIMER (Anderson et al., 2008).
The remaining analyses were performed with the statistical package
SPSS v. 24.
et al., 2012; Magni et al., 2015). For specimens with a capitulum length
3–12.45 mm we applied a growth rate of 0.624 mm/day, and for those
with a capitulum length > 12.45 mm a growth rate of 0.141 mm/day,
according to values obtained for L. anserifera (Inatsuchi et al., 2010).
Therefore, the age for, e.g., an individual of L. hillii with a capitulum of
18 mm long would be 5 + (12.45–3)/0.624 + (18–12.45)/
0.141 = 59.5 days. Note that the range of water temperature in the
study area (13.7–26.4 °C) was lower than that for which growth rates of
C. testudinaria (21.6–27.2 °C) and Lepas anserifera (23.3–29.4 °C) were
obtained (Doell et al., 2017; Inatsuchi et al., 2010). Since growth rates
of barnacles increase with temperature (Inatsuchi et al., 2010), the age
of C. testudinaria and L. hillii is likely somewhat underestimated in our
study.
Finally, we investigated whether individuals of L. hillii, C. virgatum
and C. testudinaria were attached to turtles pre- or postmortem as fol-
lows. Firstly, we calculated the postmortem interval (PMI) of each
turtle carcass according to its standard decomposition code and
monthly surface water temperature (as a proxy for sea temperature
during decomposition). Monthly temperatures from the nearest 6
sampling points to the study area from 2004 to 2017 were gathered
from Puertos.es (http://www.puertos.es/en-us) and averaged. PMIs
could then be estimated based on Fig. 4 in Santos et al. (2018), who
experimentally determined, for several sea turtle species, the minimum
days required to reach a certain decomposition code in a range of water
temperatures from ~17.0 °C to 28.7 °C. The inferred PMI for tempera-
ture and decomposition code is shown in Appendix 1. We considered
that a barnacle had attached postmortem if its estimated age was <
PMI.

2.3. Barnacle age estimation 3. Results

We estimated the age of the three potential-indicator species, i.e.,
Chelonibia testudinaria, Conchoderma virgatum and Lepas hillii (see
Results). To estimate the age of C. testudinaria, we used von
Bertalanffy’s growth equation for rostro-carinal length provided by
Doell et al. (2017), which renders age values comparable to those re-
ported by Ewers-Saucedo et al. (2015). The age for lepadomorph bar-
nacles, i.e., C. virgatum and L. hillii, was inferred from capitulum length.
For C. virgatum, we used von Bertalanffy’s growth equation provided by
Dalley and Crisp (1981). This equation was unavailable for any Lepas
spp., and we therefore estimated the age of L. hillii based on data from
allied Lepas spp., all of which exhibit comparable growth rates (see,
e.g., Evans, 1958; Skerman, 1958). Specimens of L. hillii with a capi-
tulum length ≤3 mm were assumed to be newly metamorphosed in-
dividuals with an estimated age of 5 days (Inatsuchi et al., 2010; Høeg
3.1. Description of barnacle assemblages
A total of 16,652 barnacles belonging to 13 species were collected
from the skin and carapace of 148 (74%) out of 200 loggerhead sea
turtles (Table 1). Mean abundance (95% CI) of barnacles per turtle was
81.5 (56.8–117.0). Five and 8 species were considered as obligate and
facultative commensals, respectively (Table 1). Four species exhibited a
frequency of occurrence > 5%, including two balanomorphs (Platylepas
hexastylos and C. testudinaria) and two lepadomorphs (L. hillii and C.
virgatum) (Table 1). Lepas hillii and C. virgatum significantly co-occurred
in the same turtles, and their abundances were also positively corre-
lated (Table 2). In contrast, the abundance of C. testudinaria negatively
correlated, and its presence was negatively associated, with those from
the other three species (Table 2). No other pairwise association between

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S. Ten, et al. Ecological Indicators 107 (2019) 105672

Table 2 Table 4
Associations between the four most frequent epibiont species found in logger- SIMPER analysis of the dissimilarity of barnacle assemblages between logger-
head sea turtles, Caretta caretta, from the western Mediterranean. Figures above head sea turtles, Caretta caretta, incidentally captured by pelagic longline and
the diagonal are Spearman correlation coefficients between abundances (with neritic fisheries, or turtles having only pelagic-oceanic prey (oceanic), only
‘*’ for significant values) and probabilities associated to Fisher tests [in neritic-benthic prey (neritic), or a mixed diet with both types of prey. Only
brackets]. Values below the diagonal are Spearman correlation coefficients barnacle species contributing up to c. 90% of dissimilarity are included.
between average body size of barnacles per turtle (with ‘*’ for significant va-
Species Mean abundance (four-root transformed) % Contribution to
lues). In the latter analysis, correlations involving C. testudinaria were not cal-
dissimilarity
culated due to small number of co-occurrences with the other species. ‘n’ is the
number of turtles used in each pairwise test. Fishery
Longline (N = 38) Neritic fisheries (N = 28)
P. hexastylos L. hillii C. virgatum C. testudinaria
L. hillii 2.05 0.38 37.4
P. hexastylos 1.37 1.24 22.8
P. hexastylos – 0.002 [0.424] 0.012 [1] −0.343** [0.0002]
C. virgatum 1.01 0.11 17.9
n = 137 n = 129 n = 131
C. testudinaria 0.07 0.69 12.8
L. hillii 0.163 – 0.502* [ < 0.001] −0.593** [0.063]
Total 90.0
n = 45 n = 77 n = 84
C. virgatum 0.378* 0.715** – −0. 783** [0.070] Diet
n = 29 n = 32 n = 54 Oceanic (N = 14) Neritic (N = 11)
L. hillii 3.15 0.97 37.3
‘*’ for p < 0.05 and ‘**’ for p < 0.001. P. hexastylos 1.94 1.42 26.0
C. virgatum 1.69 0.35 22.4
C. testudinaria 0 0.45 7.0
species was significant.
TOTAL 92.7
Values of capitulum length (for lepadomorph barnacles) and rostro-
Oceanic (N = 14) Mixed (N = 38)
carinal length (for balanomorph barnacles) are shown in Table 3. Le-
L. hillii 3.15 1.06 45.6
padomorph species had capituli of comparable sizes; the smallest and C. virgatum 1.69 0.39 25.1
largest average values corresponded to L. pectinata (3.12 mm) and L. P. hexastylos 1.94 1.34 23.0
anatifera (7.10 mm), whereas the smallest and largest absolute values TOTAL 93.7
corresponded to L. hillii (0.43 and 24.62 mm, respectively). Among Neritic (N = 11) Mixed (N = 38)
balanomorphs, A. amphitrite (4.69 mm) and C. caretta (37.36 mm) had P. hexastylos 1.42 1.34 33.6
the smallest and largest mean rostro-carinal length, respectively, L. hillii 0.97 1.06 30.0
C. testudinaria 0.45 0.18 13.8
whereas the smallest individual corresponded to P. hexastylos
C. virgatum 0.35 0.39 11.6
(0.66 mm), and the largest, to C. testudinaria (58.15 mm) (Table 3). TOTAL 89.0
We found a highly significant positive covariation in the average
barnacle size per turtle between L. hillii and C. virgatum; the correlation
was marginally significant for P. hexastylos and C. virgatum, and non-
significant for P. hexastylos and L. hillii (Table 2). The small number of The PERMANCOVA for the subsample of turtles incidentally cap-
co-occurrences between C. testudinaria and the other species prevented tured with at least one barnacle (n = 66) revealed significant effects of
us from estimating the correlation between the average size of C. tes- fishery (F(1, 57) = 7.815, p = 0.0001) and CCL (F(1, 57) = 4.623,
tudinaria and the other three barnacle species. p = 0.0042), and a significant ‘CCL*season’ interaction (F(3,
57) = 2.291, p = 0.0149). The SIMPER analysis for ‘fishery’ indicated
that four species were responsible for > 90% of dissimilarity between
3.2. Ecological patterns turtle samples (Table 4). Lepas hillii and C. virgatum were far more
abundant in turtles caught by pelagic longline; the abundance of P.
Results from the GLM indicated that the probability that turtles hexastylos was just slightly higher, and that of C. testudinaria was clearly
harbour barnacles was not significantly affected by CCL, season or higher in turtles caught by neritic fisheries. Mann-Whitney tests con-
provenance (p > 0.503 in all cases). None of the two-way interactions firmed these differences for individual species (p < 0.006) except for
was significant (p > 0.551 in all cases). P. hexastylos (p = 0.411). The only significant correlation between CCL
and barnacle abundance was that of L. hillii (rs = 0.262, n = 66,
Table 3 p = 0.033); correlation coefficients for the other species ranged from
Rostro-carinal length (for balanomorphs) and capitulum length (for lepado- −0.130 to 0.194, with p > 0.119 in all cases. The interaction
morphs) of epibiotic barnacles collected from 200 loggerhead turtles, Caretta
‘CCL*season’ is visually represented in Appendix 2. Note that CCL
caretta, which were incidentally captured or found stranded in the western
varied with both season and fishing gear, and some correlations be-
Mediterranean between 1990 and 2017. Measurements are expressed in mm.
tween CCL and barnacle abundance were positive or negative de-
Species Mean size ± SD Range N pending on season.
Balanomorphs
In the PERMANCOVA for the subsample of turtles with dietary data
Platylepas hexastylos 5.45 ± 3.67 0.66–23.42 6117 and harbouring at least one barnacle (n = 63), we found significant
Chelonibia testudinaria 21.63 ± 12.95 2.54–58.15 1042 effects of diet (F(2, 56) = 2.807, p = 0.0144) and season (F(3,
Chelonibia caretta 37.36 ± 8.36 28.62–45.29 3
56) = 3.366, p = 0.0008), but not of CCL (F(1, 56) = 2.041, p = 0.1021),
Stomatolepas elegans 7.04 ± 0.28 6.87–7.23 2
on barnacle assemblages. Interactions between predictors were not
Stephanolepas muricata 7.28 ± 1.61 5.67–10.03 14
Amphibalanus amphitrite 4.69 ± 1.23 3.33–6.31 10 significant (p ≫ 0.05). The post hoc analysis for ‘diet’ indicated that the
Perforatus perforatus 7.77 ± 2.84 5.25–9.78 9 barnacle assemblages of turtles with only oceanic prey (‘oceanic’ tur-
Balanus trigonus 8.10 ± 1.42 6.12–10.20 9 tles) differed from those of turtles with only neritic prey (‘neritic’ tur-
Lepadomorphs tles) (p = 0.045), or from those with neritic plus oceanic prey (‘mixed
Lepas hillii 3.87 ± 3.54 0.43–24.62 7907 diet’ turtles) (p = 0.012). According to the SIMPER analysis, four spe-
Lepas anatifera 7.10 ± 6.20 1.31–20.43 22 cies contributed to ~90% of dissimilarity: the abundance of L. hillii and
Lepas anserifera 6.62 – 1
C. virgatum was much higher in ‘oceanic’ turtles compared with ‘neritic’
Lepas pectinata 3.12 ± 2.08 0.78–8.71 33
Conchoderma virgatum 5.71 ± 3.20 1.18–19.76 1131 or ‘mixed diet’ turtles, and slightly higher for ‘mixed diet’ turtles
compared with ‘neritic’ turtles (Table 4). In contrast, the abundance of

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S. Ten, et al. Ecological Indicators 107 (2019) 105672

C. testudinaria was higher in ‘neritic’ turtles compared with the other

two categories. Differences for P. hexastylos between turtle categories
were modest, with a slight predominance in ‘oceanic’ turtles (Table 4).
Kruskall-Wallis tests confirmed a significant effect of ‘diet’ only for the
abundance of L. hillii (χ2 = 13,438, 2 df, p = 0.001) and C. virgatum
(χ2 = 11,530, 2 df, p = 0.003); the post hoc comparison confirmed the
pattern of significant differences found in the PERMANCOVA.
With regard to the seasonal effects, the post hoc analysis revealed
significant differences in autumn vs. summer (p = 0.004), autumn vs.
spring (p = 0.014) and summer vs. spring (p = 0.013). Kruskall-Wallis
tests showed significant seasonal effects for three species: L. hillii
(χ2 = 16.527, 3 df, p = 0.001; the abundance was significantly higher
in summer compared to spring); C. virgatum (χ2 = 13.425, 3 df,
p = 0.004; the abundance was significantly higher in summer com-
pared to spring), and C. testudinaria (χ2 = 13,216, 3 df, p = 0.004; the
abundance was significantly higher in autumn compared with the re-
maining seasons) (see also Appendix 3).
A total of 85.4% of turtles (80.5% after cross-validation) were cor-
rectly allocated to fishery using CDA based on data from turtles har-
bouring C. testudinaria and/or the lepadormorph barnacles L. hillii and
C. virgatum (n = 41; Appendix 4). The actual number of turtles captured
by longline and neritic fisheries were 32 (78%) and 9 (22%), whereas
the predicted numbers were 34 (82.9%) and 7 (17.1%). When the
discriminant function was applied to stranded turtles harbouring at
least one of these indicator barnacle species (n = 52), a total of 44
(84.6%) were assigned to longline fisheries and 8 (15.4%) to neritic
fisheries (Appendix 4). The proportion of turtles in each category was
similar to that observed for captured turtles (Fisher test, p = 0.433).

3.3. Inferences based on barnacle age estimations

The estimated age of barnacle species per turtle and age distribution
per species are shown in Table 5 and Fig. 1. On average, individuals of
C. testudinaria were older than those of C. virgatum and L. hillii (months
vs. days), showing a clearly bimodal age distribution (Fig. 1). The oldest
individual of C. testudinaria was estimated to be 1118 days old (about
the maximum recorded age for this species; Ewers-Saucedo et al.,
2015), and the youngest, 18 days old. We inferred that all specimens of
C. testudinaria had attached premortem (Table 5). The youngest and
oldest individuals of L. hillii were 5 and 214 days old. There was a vast
predominance of young individuals (< 10 days old), but the age dis-
tribution exhibited a second (modest) peak in the range of 40–70 days

Table 5
Average value ± SD [median] for the minimum, median, and maximum age
per turtle (days) of the epibiotic barnacles Lepas hillii, Conchoderma virgatum
and Chelonibia testudinaria collected from loggerhead sea turtles, Caretta caretta,
in the western Mediterranean. The moment of attachment (pre-mortem, post-
mortem or both) was determined by comparing the minimum and maximum
ages of each barnacle species per turtle with the estimated time since the turtle
died according to its degree of decomposition (see the text for details). Only
turtles with known decomposition codes were included in the analysis.
C. virgatum L. hillii C. testudinaria

Age (days) Fig. 1. Age distribution of the indicator barnacle species (a) Lepas hillii, (b)
n turtles 31 71 13 Conchoderma virgatum and (c) Chelonibia testudinaria collected from a sample of
loggerhead sea turtles, Caretta caretta, from the western Mediterranean. This
Minimum 4.8 ± 4.2 [3.4] 9.1 ± 20.0 [5.0] 172.9 ± 263.4 [88.4] figure was performed with RStudio.
Median 8.4 ± 5.7 [7.5] 13.1 ± 24.3 [6.0] 275.9 ± 305.3 [119.5]
Maximum 13.6 ± 6.6 [13.2] 33.4 ± 44.4 [13.7] 495.8 ± 388.9 [386.9]
(Fig. 1). In contrast, the sample of C. virgatum displayed a unimodal
Moment of attachment
n turtles 30 57 13 distribution of individuals with a range of ages from just 1 to 30 days
old. According to the PMIs of turtles, a sizable portion of barnacles of
Pre-mortem only 13 (43.3%) 38 (66.7%) 13 (100.0%) both species, especially C. virgatum, would have been attached post-
Post-mortem 2 (6.7%) 3 (5.2%) 0 (0%) mortem (Table 5, Fig. 1).
only
Both 15 (50.0%) 16 (28.1%) 0 (0%)
Inferences on habitat use for 41 turtles based on data on occurrence
and age distribution of C. virgatum, L. hillii and C. testudinaria are shown

5
S. Ten, et al.
in Appendix 5. A total of 37 turtles were predicted to have visited
oceanic waters, with a (conservative) range of residence time from 5 to
70 days, distributed in 1–4 visits; three turtles had visited neritic waters
between 18 days and 26 months before capture or stranding; and one
turtle visited (first) neritic and (then) oceanic habitats. These inferences
were compatible with data on habitat of capture and diet for 82.9% of
turtles (Appendix 5).
4. Discussion
4.1. Barnacles as ecological indicators
Over nearly three decades, the epibiotic barnacle assemblage of
juvenile loggerhead sea turtles in the study area have consistently been
dominated by four species, i.e., the obligate commensals P. hexastylos
and C. testudinaria and the facultative commensals L. hillii and C. vir-
gatum, with other 9 barnacle species seldom occurring on the turtles
(Domènech et al., 2015b and references therein; present study). These
four taxa appear to be the most regionally frequent and have been re-
ported on other juvenile loggerhead turtles from the western (Laurent,
1988), central (Gramentz, 1988; Scaravelli et al., 2003; Casale et al.,
2012b) and eastern (Kitsos et al., 2003) Mediterranean, sometimes
being accompanied by barnacle species that are locally common (e.g.,
Stomatolepas elegans or Balanus trigonus).
Our results provide strong quantitative support that 3 of these 4
species, namely, L. hillii, C. virgatum and C. testudinaria, have potential
value as ecological indicators of habitat, as Casale et al. (2004, 2012b)
suggested. We preferentially found L. hillii and C. virgatum on turtles
that had been captured by longliners, or on turtles with dietary contents
comprising only pelagic-oceanic prey. Apparently, previous surveys
also reported lepadomorph barnacles, especially Lepas spp. and C. vir-
gatum, on loggerhead turtles captured in the open sea (Gramentz, 1988;
Laurent, 1988; Casale et al., 2004). Both C. virgatum and Lepas spp. are
cosmopolitan fouling species that inhabit warm-temperate oceanic
waters and rapidly colonize any drifting object (Skerman, 1958; Patel,
1959; Southward, 1987; Harper, 1995; Hinojosa et al., 2006; Fraser
et al., 2011; González et al., 2012; Frick and Pfaller, 2013; Schiffer and
Herbig, 2016). This ‘fugitive’ lifestyle, coupled with the likely high
abundance these species in the open ocean could explain, not only that
they strongly co-occur on the same individual turtles in our sample, but
also that their sizes co-varied significantly, which would suggest con-
joint recruitment.
In contrast, our results indicate that C. testudinaria was more asso-
ciated with turtles captured by neritic fisheries or with a diet that in-
cluded benthic-neritic prey. In agreement with this pattern, C. testudi-
naria is the most frequent and abundant epibiont species found in
coastal nesting females of both loggerhead sea turtles and green turtles
(Chelonia mydas) from the eastern Mediterranean (Fuller et al., 2010); it
also occurs frequently on juvenile turtles inhabiting shallow waters
(Scaravelli et al., 2003; Casale et al., 2004). Consequently, the abun-
dance of C. testudinaria should covary negatively with that of L. hillii
and C. virgatum, such as we observed. The preferential distribution of C.
testudinaria in neritic waters could result from two combined factors.
First, this species is able to settle, not only on sea turtles, but also on a
variety of benthic neritic organisms with hard external surfaces, in-
cluding crustaceans, gastropods and even sea snakes (Cheang et al.,
2013). Second, the development of C. testudinaria from hatchling to
cyprid is short, taking about 9 days (Zardus and Hadfield, 2004). Ac-
cordingly, the production of larval stages should largely occur in neritic
waters, and dispersal to open waters should be restricted by the short
larval lifespan. Although there is the possibility that adults of C. testu-
dinaria, being long-lived, reproduce and release eggs when turtle hosts
visit oceanic waters, recruitment to new hosts would be hampered
under pelagic conditions because of the ‘dilution’ effects affecting larval
stages and the low host density (see Rawson et al., 2003). We assume
that the effect of low host density is much stronger for C. testudinaria
6
Ecological Indicators 107 (2019) 105672
than for the facultative commensals L. hillii and C. virgatum, which can
attach to a great variety of floating substrata.
Casale et al. (2012b) proposed that Platylepas spp. could be typically
found on turtles exploiting pelagic habitats, although their suggestion
was based on limited evidence. In our study, the abundance of P. hex-
astylos did not significantly differ in ‘oceanic’ vs. ‘neritic’ turtles.
Nevertheless, its abundance was negatively correlated with that of C.
testudinaria, and mean body size per turtle covaried weakly, but posi-
tively, with that of one of the oceanic species, i.e., C. virgatum. Although
the latter two observations lend support to Casale et al. (2012b)’s hy-
pothesis, P. hexastylos showed limited value as a habitat indicator.
Age estimation adds a timescale for the residence of indicator bar-
nacles, hence their hosts/substrata, in a given habitat (e.g., Fraser et al.,
2011; Sloan et al., 2014). In the present study, we are aware that
barnacle age estimates were coarse for three reasons: (i) growth rates
obtained for barnacles are affected by many biotic and abiotic factors
(see, e.g., Burrows et al., 2010; Inatsuchi et al., 2010); (ii) for two
species, we used growth rates obtained at higher temperatures that
those recorded in the study area, and (iii) for one species, we used
surrogate values gathered from allied species. Notwithstanding, the
inferences drawn on the time since barnacle settlement, or on habitat
exploitation by turtles, were largely congruent with the evidence ob-
tained from diet and fishery interactions, and are likely valid for the
interpretation purposes we deal with below.
Lepadomorph barnacles, being fast-growing and early-maturing
species, have been used to estimate floating times of objects at the scale
of ca. 1–8 weeks (Fraser et al., 2011; Magni et al., 2015; López et al.,
2017). In our study, the estimated median age for C. virgatum was just
about one week and, in half of turtles harbouring this species, at least
some individuals were presumably attached postmortem. Regardless of
precise figures, we interpret that C. virgatum could function as a good
indicator of the habitat where a turtle was shortly before, or after,
death. However, the samples of L. hillii were composed of compara-
tively older individuals (range of median ages: 9–33 days), thus ex-
panding information on habitat use by turtles up to a few weeks before
collection. Interestingly, the residence time in oceanic waters inferred
from the age of L. hillii was from 5 to 70 days, with most values
being < 60 days (Appendix 5). These values are compatible with data
on residence times obtained for satellite-tracked turtles in the western
Mediterranean (Cardona et al., 2005; Revellés et al., 2007b). Finally,
specimens of C. testudinaria were on average much older (several
months), and showed higher age variability, than lepadomorph bar-
nacles. Consequently, C. testudinaria could tag mainly the visits (of
unknown duration) of turtles to neritic habitats. It is interesting to note
that none of the individuals of C. testudinaria attached post-mortem.
One could argue that this is unsurprising because C. testudinaria is an
obligate epibiont; however, recent experimental data have proven that
this species can settle on detached scutes of loggerheads (Sloan et al.,
2014). The absence of newly recruited individuals could therefore be
related, at least in part, to a low density of larval stages in the study
area and/or a short residence time of juvenile turtles in neritic waters.
Two observations conform to this hypothesis: (i) the prevalence of C.
testudinaria in our sample was low and (ii) data on satellite-tracked
juvenile turtles in the western Mediterranean indicate that many of
them spend little time in neritic waters (Cardona et al., 2005; Revellés
et al., 2007b; but see Cardona et al., 2009 for some exceptions).
4.2. Confounding factors
In the PERMANCOVA of turtles that were incidentally captured, we
detected a significant effect of CCL, and of the ‘CCL*season’ interaction,
on barnacle assemblages. However, the effect of CCL vanished and a
‘pure’ seasonal effect showed up in the analysis involving turtles with
dietary data. The interpretation of these patterns is difficult because of
the heterogeneity of the sample. In particular, the largest turtles were
collected in the summer (June-September) by longliners in oceanic
S. Ten, et al.
waters (Appendix 2) and, to a lesser extent; large turtles also had a diet
composed of oceanic prey (Appendix 3). Accordingly, we would expect
(and we actually observed) that large turtles collected in the summer
harbour larger loads of C. virgatum and L. hillii because these turtles
were in pelagic-oceanic waters. Therefore, the significant effects of CCL
and season we detected could largely result from pooling turtle samples
from different seasons, which was also reflected in C. virgatum and L.
hillii abundances.
However, one could argue that turtles captured by longliners, or
with a pelagic-oceanic diet, actually harboured more individuals of C.
virgatum and L. hillii because these turtles tended to be larger and/or be
collected in the summer (instead of these turtles being more “oceanic”).
In theory, larger turtles provide more surface of contact for barnacle
larvae, but this size effect of substratum could be detected only if sur-
face variability is large and/or the rate of barnacle recruitment is high
(see, e.g., Gil and Pfaller, 2016). Furthermore, the size effect should be
positive for all barnacle species. None of these conditions are fulfilled in
our case: the CCL of most turtles was in a narrow range (~40–80 cm),
average abundances of barnacle ranged from just 5 to 40 individuals
per turtle (thus implying low recruitment rates), and correlation of CCL
and abundance was negative for C. testudinaria. With regard to seasonal
effects, it is known that the breeding cycle of barnacles may differ
widely even in species of the same genus (Barnes, 1989). Although
there is no specific information for C. virgatum or L. hillii, the limited
data available for other Lepas spp. in the Mediterranean indicate re-
productive periods in spring or throughout the year (Barnes, 1989).
In summary, we cannot rule out that size-related or seasonal effects
may influence recruitment patterns of the barnacles here studied, but
our data suggest that these potential effects are not confounding the
value of these barnacles as indicators of habitat use by turtles.
4.3. Practical implications
To increase knowledge on sea turtle movements and habitat use, we
firstly encourage researchers worldwide to identify and count in-
dividuals of C. testudinaria, C. virgatum and L. hillii (and other Lepas
spp.) on live turtles, or on turtle carcasses, that are found stranded for
unknown reasons. These indicator barnacles demonstrated a significant
ability to predict the habitat of incidental capture of loggerhead turtles
using CDA (over 80% of success after cross validation), with predictive
success being similar for oceanic and neritic gears. More importantly,
when we applied the discriminant function to a sample of stranded
turtles of unknown origin, the proportions of turtles assigned to puta-
tive longline fisheries and neritic fisheries were surprisingly similar to
those observed in the sample of captured turtles. Obviously, this ana-
lysis assumes that stranded turtles had also interacted with fisheries,
which is not unrealistic: fishery interactions is, by far, the most frequent
cause of mortality for loggerhead sea turtles in the western
Mediterranean (Aguilar et al., 1995; Carreras et al., 2004; Camiñas
et al., 2006; Tomás et al., 2008).
Secondly, we recommend researchers to measure the size, ideally of
all individuals of indicator species, or at least of the smallest and largest
individual of each species. Size data can be converted to crude age
estimations that are highly informative. On the one hand, the presence
of small (=young) individuals of any of these barnacle species on turtle
carcasses may indicate the habitat where the turtle died and/or drifted
afterwards, and for how long (see Magni et al., 2015). This forensic
information is especially useful for stranded carcasses, particularly if
combined with other sources of evidence (diet, pathology, etc.). On the
other hand, large (=old) barnacles may assist in tracing the use of
different habitats by live turtles. For instance, one of the loggerheads
included in this study (CCL: 74 cm) harboured individuals of both C.
testudinaria (range of estimated ages: 19–34 months) and L. hillii (range
of estimated ages: 5–60 days). Based on these data, we could infer that
this turtle visited neritic waters ca. 3 years before death, moved to
oceanic waters at least two months before death, and died there (as the
7
Ecological Indicators 107 (2019) 105672
signs of interaction with longliners confirmed).
In conclusion, epibiotic barnacles are a cost-effective tool to assist in
investigating patterns of habitat use by marine turtles, which are all
threatened and highly vagile species. Information derived from barna-
cles along with other sources of evidence will allow implementing
geographically-tailored conservation strategies to mitigate the impact
of fisheries.
Acknowledgements
We thank the Wildlife Service and the Conselleria d'Agricultura,
Desenvolupament Rural, Emergència Climàtica i Transició Ecològica of
the Generalitat Valenciana for logistic and financial support. Financial
support was also given by the project PROMETEOII/2015/018 of the
Conselleria d'Educació, Investigació, Cultura i Esport of the Generalitat
Valenciana. F. Domènech benefits from a predoctoral grant UV-INV-
PREDOC15-265927. The assistance of the staff of the Marine Zoology
Unit, especially Francisco Javier Badillo, and the Oceanogràfic aqua-
rium of Valencia is much appreciated.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.ecolind.2019.105672.
References
Abalo-Morla, S., Marco, A., Tomás, J., Revuelta, O., Abella, E., Lorenzo, T., Crespo-Picazo,
J.L., Fernández, C., Valdés, F., Arroyo, M.C., Montero, S., Vázquez, C., Eymar, J.,
Esteban, J.A., Pelegrí, J., Belda, E.J., 2018. Survival and dispersal routes of head-
started post-hatchlings of loggerhead sea turtle (Caretta caretta) in the Mediterranean
Sea. Mar. Biol. 165, 5. https://doi.org/10.1007/s00227-018-3306-2.
Aguilar, R., Mas, J., Pastor, X., 1995. Impact of the Spanish swordfish longline fisheries on
the loggerhead sea turtle Caretta caretta population in the western Mediterranean.
12th Annual Workshop on Sea Turtle Biology and Conservation, Jekyll Island,
Georgia, EE.UU.
Álvarez de Quevedo, I., de Haro, A., Pubill, E., Cardona, L., Aguilar, A., 2006. Bottom
trawling is a threat for the conservation of loggerhead sea turtles off north-eastern
Spain. In: Frick, M., Panagopoulou, A., Rees, A.F., Williams, K. (Eds.), 26th Annual
Symposium on Sea Turtle Biology and Conservation, Athens.
Anderson, M.J., Gorley, R.N., Clarke, R.K., 2008. Permanova+ for Primer: Guide to
Software and Statistical Methods. PRIMER-E Ltd, Plymouth, United Kingdom.
Báez, J.C., García-Barcelona, S., Camiñas, J.A., Macías, D., 2019. Fishery strategy affects
the loggerhead sea turtle mortality trend due to the longline bycatch. Fish. Res. 212,
21–28. https://doi.org/10.1016/j.fishres.2018.11.032.
Báez, J.C., Macías, D., Camiñas, J.A., Ortiz De Urbina, J.M., García-Barcelona, S., Bellido,
J.J., Real, R., 2013. Bycatch frequency and size differentiation in loggerhead turtles
as a function of surface longline gear type in the western Mediterranean Sea. J. Mar.
Biol. Assoc. U.K. 93, 1423–1427.
Báez, J.C., Real, R., Camiñas, J.A., 2007a. Differential distribution within longline
transects of loggerhead turtles and swordfish captured by the Spanish Mediterranean
surface longline fishery. J. Mar. Biol. Assoc. U.K. 87 (3), 801–803.
Báez, J.C., Real, R., García-Soto, C., de la Serna, J., Macías, D., Camiñas, J.A., 2007b.
Loggerhead turtle by-catch depends on distance to the coast, independent of fishing
effort: implications for conservation and fisheries management. Mar. Ecol. Prog. Ser.
338, 249–256.
Barnes, M., 1989. Egg production in cirripedes. Oceanogr. Mar. Biol. Annu. Rev. 27,
91–166.
Bolten, A.B., 2003. Variation in sea turtle life history patterns: neritic vs. oceanic de-
velopmental stages. In: In: Lutz, P.L., Musick, J., Wyneken, J. (Eds.), The Biology of
Sea Turtles, volume II. CRC Press, Boca Raton, FL, pp. 243–257.
Burrows, M.T., Jenkins, S.R., Robb, L., Harvey, R., 2010. Spatial variation in size and
density of adult and post-settlement Semibalanus balanoides: effects of oceano-
graphic and local conditions. Mar. Ecol. Prog. Ser. 398, 207–219.
Camiñas, J.A., Báez, J.C., Valeiras, X., Real, R., 2006. Differential loggerhead bycatch and
direct mortality due to surface longlines according to boat strata and gear type. Sci.
Mar. 70, 661–665.
Cardona, L., Revellés, M., Carreras, C., San Félix, M., Gazo, M., Aguilar, A., 2005. Western
Mediterranean immature loggerhead turtles: habitat use in spring and summer as-
sessed through satellite tracking and aerial surveys. Mar. Biol. 147, 583–591.
Cardona, L., Revellés, M., Parga, M.L., Tomás, J., Aguilar, A., Alegre, F., Raga, A., Ferrer,
X., 2009. Habitat use by loggerhead sea turtles Caretta caretta off the coast of eastern
Spain results in a high vulnerability to neritic fishing gear. Mar. Biol. 156,
2621–2630. https://doi.org/10.1007/s00227-009-1288-9.
Carreras, C., Cardona, L., Aguilar, A., 2004. Incidental catch of the loggerhead turtle
Caretta caretta off the Balearic Islands (western Mediterranean). Biol. Conserv. 117,
321–329.
S. Ten, et al.
Carreras, C., Pascual, M., Cardona, L., Marco, A., Bellido, J., Castillo, J., Tomás, J., Raga,
J., Sanfélix, M., Fernández, G., Aguilar, A., 2011. Living Together but Remaining
Apart: Atlantic and Mediterranean Loggerhead Sea Turtles (Caretta caretta) in Shared
Feeding Grounds. J. Heredity 102 (6), 666–677.
Casale, P., Abbate, G., Freggi, D., Conte, N., Oliverio, M., Argano, R., 2008. Foraging
ecology of loggerhead sea turtles Caretta caretta in the central Mediterranean Sea:
evidence for a relaxed life history model. Mar. Ecol. Prog. Ser. 372, 265–276.
Casale, P., Broderick, A.C., Freggi, D., Mencacci, R., Fuller, W.J., Godley, B.J., Luschi, P.,
2012a. Long-term residence of juvenile loggerhead turtles to foraging grounds: a
potential conservation hotspot in the Mediterranean. Aquat. Conserv. Mar.
Freshwater Ecosyst. 22, 144–154. https://doi.org/10.1002/aqc.2222.
Casale, P., D’Addario, M., Freggi, D., Argano, R., 2012b. Barnacles (Cirripedia, Thoracica)
and associated epibionts from sea turtles in the central Mediterranean. Crustaceana
85, 533–549.
Casale, P., Freggi, D., Basso, R., Argano, R., 2004. Epibiotic barnacles and crabs as in-
dicators of Caretta caretta distribution and movements in the Mediterranean Sea. J.
Mar. Biol. Assoc. U.K. 84, 1005–1006.
Casale, P., Mazaris, A., Freggi, D., 2011. Estimation of age at maturity of loggerhead sea
turtles Caretta caretta in the Mediterranean using length-frequency data. Endanger
Species Res. 13 (2), 123–129.
Cheang, C., Tsang, L., Chu, K., Cheng, I., Chan, B., 2013. Host-specific phenotypic plas-
ticity of the turtle barnacle Chelonibia Testudinaria: a widespread generalist rather
than a specialist. PLoS One 8, e57592.
Clarke, K.R., Gorley, R.N., 2006. Primer v6: User Manual/Tutorial. PRIMER-E Ltd,
Plymouth, United Kingdom.
Clusa, M., Carreras, C., Pascual, M., Gaughrana, S.J., Piovano, S., Fernández, G., Tomás,
J., Giacoma, C., Raga, J.A., Aguilar, A., Cardona, L., 2016. Potential bycatch impact
on distinct sea turtle populations is dependent on fishing ground rather than gear
type in the Mediterranean Sea. Mar. Biol. 163, 1–10. https://doi.org/10.1007/
s00227-016-2875-1.
Clusa, M., Carreras, C., Pascual, M., Gaughran, S.J., Piovano, S., Giacoma, C., Fernández,
G., Levy, Y., Tomás, J., Raga, J.A., Maffucci, F., Hochscheid, S., Aguilar, A., Cardona,
L., 2014. Fine-scale distribution of juvenile Atlantic and Mediterranean loggerhead
turtles (Caretta caretta) in the Mediterranean Sea. Mar. Biol. 161, 509–519. https://
doi.org/10.1007/s00227-013-2353-y.
Dalley, R., Crisp, D.J., 1981. Conchoderma: a fouling hazard to ships underway. Mar.
Biol. Lett. 2, 141–152.
Di Bello, A., Valastro, C., Staffieri, F., Crovace, A., 2006. Contrast radiography of the
gastrointestinal tract in sea turtles. Vet. Radiol. Ultrasound 47 (4), 351–354.
Doell, S., Connolly, R., Limpus, C., Pearson, R., van de Merwe, J., 2017. Using growth
rates to estimate age of the sea turtle barnacle Chelonibia testudinaria. Mar. Biol.
164, 222.
Domènech, F., Álvarez de Quevedo, I., Merchán, M., Revuelta, O., Vélez-Rubio, G., Bitón,
S., Cardona, L., Tomás, J., 2015a. Incidental catch of marine turtles by Spanish
bottom trawlers in the western Mediterranean. Aquat. Conserv. Mar. Freshwater
Ecosyst. 25, 678–689. https://doi.org/10.1002/aqc.2463.
Domènech, F., Badillo, F.J., Tomás, J., Raga, J.A., Aznar, F.J., 2015b. Epibiont commu-
nities of loggerhead marine turtles (Caretta caretta) in the western Mediterranean:
influence of geographic and ecological factors. J. Mar. Biol. Assoc. U.K. 95, 851–861.
Eckert, S.A., Moore, J.E., Dunn, D.C., Van Buiten, R.S., Eckert, K.L., Halpin, P.N., 2008.
Modeling loggerhead turtle movement in the Mediterranean: importance of body size
and oceanography. Ecol. Appl. 18 (2), 290–308.
Engqvist, L., 2005. The mistreatmen of covariable interaction terms in linear model
analyses of behavioural and evolutionary ecology studies. Anim. Behav. 70, 967–971.
Evans, F., 1958. Growth and maturity of the barnacles Lepas hillii and Lepas anatifera.
Nature 182, 1245–1246.
Ewers-Saucedo, C., Arendt, M.D., Wares, J.P., Rittschof, D., 2015. Growth, mortality, and
mating group size of an androdioecious barnacle: implications for the evolution of
dwarf males. J. Crustac. Biol. 35, 166–176.
Fraser, C., Nikula, R., Waters, J., 2011. Oceanic rafting by a coastal community. Proc. R.
Soc. Lond. 278 (1706), 649–655. https://doi.org/10.1098/rspb.2010.1117.
Frick, M.G., Pfaller, J.B., 2013. Sea turtle epibiosis. In: In: Wyneken, J., Lohmann, K.J.,
Musick, J.A. (Eds.), The Biology of Sea Turtles, vol. 3. CRC Press, Boca Ratón, pp.
399–426.
Fuller, W., Broderick, A., Enever, R., Thorne, P., Godley, B., 2010. Motile homes: a
comparison of the spatial distribution of epibiont communities on Mediterranean Sea
turtles. J. Nat. Hist. 44 (27–28), 1743–1753.
Gibbons, M., Richardson, A., 2008. Patterns of jellyfish abundance in the North Atlantic.
Hydrobiologia 616 (1), 51–65. https://doi.org/10.1007/s10750-008-9593-8.
Gil, M., Pfaller, J., 2016. Oceanic barnacles act as foundation species on plastic debris:
implications for marine dispersal. Sci. Rep. 6 (1).
Goatley, C., Hoey, A., Bellwood, D., 2012. The role of turtles as coral reef macro-
herbivores. PLoS One 7 (6), e39979. https://doi.org/10.1371/journal.pone.0039979.
Gómez de Segura, A., Tomás, J., Pedraza, S.N., Crespo, E.A., Raga, J.A., 2006. Abundance
and distribution of the endangered loggerhead turtle in Spanish Mediterranean wa-
ters and the conservation implications. Anim. Conserv. 9, 199–206.
González, J., Martín, L., Herrera, R., González-Lorenzo, G., Espino, F., Barquín-Diez, J.,
Southward, A., 2012. Cirripedia of the Canary Islands: distribution and ecological
notes. J. Mar. Biol. Assoc. U.K. 92 (1), 129–141.
Gramentz, D., 1988. Prevalent epibiont sites on Caretta caretta in the Mediterranean Sea.
Naturalista sicil 12 (1–2), 33–46.
Hamann, M., Godfrey, M., Seminoff, J., Arthur, K., Barata, P., Bjorndal, K., Bolten, A.,
Broderick, A., Campbell, L., Carreras, C., Casale, P., Chaloupka, M., Chan, S., Coyne,
M., Crowder, L., Diez, C., Dutton, P., Epperly, S., FitzSimmons, N., Formia, A.,
Girondot, M., Hays, G., Cheng, I., Kaska, Y., Lewison, R., Mortimer, J., Nichols, W.,
Reina, R., Shanker, K., Spotila, J., Tomás, J., Wallace, B., Work, T., Zbinden, J.,
8
Ecological Indicators 107 (2019) 105672
Godley, B., 2010. Global research priorities for sea turtles: informing management
and conservation in the 21st century. Endanger Species Res. 11 (3), 245–269.
Hannan, L., Roth, J., Ehrhart, L., Weishampel, J., 2007. Dune vegetation fertilization by
nesting sea turtles. Ecology 88 (4), 1053–1058. https://doi.org/10.1890/06-0629.
Harper, D.E., 1995. Fouling of towed seismic streamers off central Africa by the lepa-
domorph barnacle Conchoderma Virgatum. Crustaceana 68 (6), 779–781. https://
doi.org/10.1163/156854095x00287.
Hayashi, R., 2009. New host records of the turtle barnacle, Cylindrolepas sinica: a case
study of sea turtles’ behaviour and their epibionts. Mar. Biodivers. Rec. 2, e165.
https://doi.org/10.1017/S1755267209990947.
Hearne, E., Johnson, R., Gulick, A., Candelmo, A., Bolten, A., Bjorndal, K., 2019. Effects of
green turtle grazing on seagrass and macroalgae diversity vary spatially among
seagrass meadows. Aquat. Bot. 152, 10–15. https://doi.org/10.1016/j.aquabot.2018.
09.005.
Hinojosa, I., Boltana, S., Lancellotti, D., Macaya, E., Ugalde, P., Valdivia, N., Vasquez, N.,
Newman, W.A., Thiel, M., 2006. Geographic distribution and description of four
pelagic barnacles along the south east Pacific coast of Chile – a zoogeographical
approximation. Rev. Chil. Hist. Nat. 79, 13–27.
Høeg, J.T., Maruzzo, D., Okano, K., Glenner, H., Chan, B.K., 2012. Metamorphosis in
balanomorphan, pedunculated, and parasitic barnacles: a video-based analysis.
Integr. Comp. Biol. 52, 337–347.
Inatsuchi, A., Yamato, S., Yusa, Y., 2010. Effects of temperature and food availability on
growth and reproduction in the neustonic pedunculate barnacle Lepas anserifera.
Mar. Biol. 157, 899–905.
IUCN-SSC Marine Turtle Specialist Group. 2019. Red List. [online] Available at: https://
iucn-mtsg.org/about/structure-role/red-list/.
Kitsos, M., Christodoulou, M., Kalpakis, S., Noidou, M., Koukouras, A., 2003. Cirripedia
Thoracica associated with Caretta caretta (Linnaeus, 1758) in the northern Aegean
Sea. Crustaceana 76 (4), 403–409.
Koch, A.U., Willan, C.C., 2007. Natator depressus (flatback sea turtle) habitat. Herp. Rev.
38, 448.
Laurent, L., 1988. Observations pélagiques de la caouanne, Caretta caretta Linnaeus
(Chelonii, Cheloniidae) en Méditerranée occidentale. Bull. Soc. Herpétol. France 45,
9–16.
López, B., Macaya, E., Tala, F., Tellier, F., Thiel, M., 2017. The variable routes of rafting:
stranding dynamics of floating bull kelp Durvillaea antarctica (Fucales,
Phaeophyceae) on beaches in the SE Pacific. J. Phycol. 53 (1), 70–84.
Magni, P.A., Venn, C., Aquila, I., Pepe, F., Ricci, P., Di Nunzio, C., Ausania, F., Dadour,
I.R., 2015. Evaluation of the floating time of a corpse found in a marine environment
using the barnacle Lepas anatifera L. (Crustacea: Cirripedia: Pedunculata). Forensic
Sci. Int. 247, e6–e10.
Monzón-Argüello, C., Rico, C., Naro-Maciel, E., Varo-Cruz, N., López, P., Marco, A.,
López-Jurado, L.F., 2010. Population structure and conservation implications for the
logger-head sea turtle of the Cape Verde Islands. Conserv. Gen. 11, 1871–1884.
Patel, B., 1959. The influence of temperature on the reproduction and moulting of Lepas
anatifera L. under laboratory conditions. J. Mar. Biol. Assoc. U.K. 38, 589–597.
Rawson, P.D., Macnamee, R., Frick, M.G., Williams, K.L., 2003. Phylogeography of the
coromulid barnacle, Chelonibia testudinaria, from loggerhead sea turtles, Caretta
caretta. Mol. Ecol. 12, 2697–2706.
Rees, A.F., Alfaro-Shigueto, J., Barata, P.C.R., Bjorndal, K.A., Bolten, A.B., Bourjea, J.,
Broderick, A.C., Campbell, L.M., Cardona, L., Carreras, C., Casale, P., Ceriani, S.A.,
Dutton, P.H., Eguchi, T., Formia, A., Fuentes, M.M.P.B., Fuller, W.J., Girondot, M.,
Godfrey, M.H., Hamann, M., Hart, K.M., Hays, G.C., Hochscheid, S., Kaska, Y.,
Jensen, M.P., Mangel, J.C., Mortimer, J.A., Naro-Maciel, E., Ng, C.K.Y., Nichols, W.J.,
Phillott, A.D., Reina, R.D., Revuelta, O., Schofield, G., Seminoff, J.A., Shanker, K.,
Tomás, J., van de Merwe, J.P., Van Houtan, K.S., Vander Zanden, H.B., Wallace, B.P.,
Wedemeyer-Strombel, K.R., Work, T.M., Godley, B.J., 2016. Are we working towards
global research priorities for management and conservation of sea turtles? Endanger
Species Res. 31, 337–382.
Reiczigel, J., 2003. Confidence intervals for the binomial parameter: some new con-
siderations. Stat. Med. 22, 611–621.
Reiczigel, J., Rozsa, L., Reiczigel, A., Fabian, I., 2013. Quantitative Parasitology (QPweb).
Available at. www2.univet.hu/qpweb.
Revellés, M., Camiñas, J.A., Cardona, L., Parga, M., Tomás, J., Aguilar, A., Alegre, F.,
Raga, A., Bertolero, A., Oliver, G., 2008. Tagging reveals limited exchange of im-
mature loggerhead sea turtles (Caretta caretta) between regions in the western
Mediterranean. Sci. Mar. 72 (3), 511–518.
Revellés, M., Cardona, L., Aguilar, A., Borrel, A., Fernández, G., San Félix, M., 2007a.
Stable C and N isotope concentration in several tissues of the loggerhead sea turtle
Caretta caretta from the western Mediterranean and dietary implications. Sci. Mar.
71 (1), 87–93.
Revellés, M., Cardona, L., Aguilar, A., San Félix, M., Fernández, G., 2007b. Habitat use by
immature loggerhead sea turtles in the Algerian Basin (western Mediterranean):
swimming behaviour, seasonality and dispersal pattern. Mar. Biol. 151, 1501–1515.
Rózsa, L., Reiczigel, J., Majoros, G., 2000. Quantifying parasites in samples of hosts. J.
Parasitol. 86, 228–232.
Santos, B., Kaplan, D., Friedrichs, M., Barco, S., Mansfield, K., Manning, J., 2018.
Consequences of drift and carcass decomposition for estimating sea turtle mortality
hotspots. Ecol. Indic. 84, 319–336.
Sazima, I., Grossman, A., Sazima, C., 2004. Hawksbill turtles visit mustached barbers:
cleaning symbiosis between Eretmochelys imbricata and the shrimp, Stenopus his-
pidus. Biota Neotrop. 4 (1), 1–6.
Scaravelli, D., Affronte, M., Costa, F., 2003. Analysis of epibiont presence on
Carettacaretta from Adriatic Sea. In: Margaritoulis, D.Y., Demetropoulos, A. (Eds.),
Proceedings of the First Mediterranean Conference on Marine Turtles, Barcelona
Convention – Bern Convention – Bonn Convention (CMS), Nicosia, pp. 221–225.
S. Ten, et al.
Schiffer, P., Herbig, H., 2016. Endorsing Darwin: global biogeography of the epipelagic
goose barnacles Lepas spp. (Cirripedia, Lepadomorpha) proves cryptic speciation.
Zool. J. Linn. Soc. 177 (3), 507–525. https://doi.org/10.1111/zoj.12373.
Schofield, G., Katselidis, K.A., Dimopoulos, P., Pantis, J.D., Hays, G.C., 2006. Behaviour
analysis of the loggerhead sea turtle Caretta caretta from direct in-water observation.
Endanger Species Res. 2, 71–79.
Skerman, T.M., 1958. Rate of growth in two species of Lepas (Cirripedia). NZ. J. Sci. 1,
402–411.
Sloan, K., Zardus, J.D., Jones, M.L., 2014. Substratum fidelity and early growth in
Chelonibia testudinaria, a turtle barnacle especially common on debilitated logger-
head (Caretta caretta) sea turtles. Bull. Mar. Sci. 90 (2), 581–597. https://doi.org/10.
5343/bms.2013.1033.
Southward, A., 1987. In: Barnacle Biology, pp. 329–360.
Stamper, M., Harms, C., Epperly, S., Braun-McNeill, J., Avens, L., Stoskopf, M., 2005.
Relationship between barnacle epibiotic load and hematologic parameters in log-
gerhead sea turtles (Caretta caretta), a comparison between migratory and residential
animals in Pamlico Sound, North Carolina. J. Zoo Wildl. Med. 36 (4), 635–641.
https://doi.org/10.1638/04-074.1.
Tomás, J., Aznar, F., Raga, J., 2001. Feeding ecology of the loggerhead turtle Caretta
caretta in the western Mediterranean. J. Zool. 255 (4), 525–532. https://doi.org/10.
1017/s0952836901001613.
Ecological Indicators 107 (2019) 105672
Tomás, J., Gozalbes, P., Raga, J., Godley, B., 2008. Bycatch of loggerhead sea turtles:
insights from 14 years of stranding data. Endanger Species Res. 5, 161–169.
Valente, A., Marco, I., Parga, M., Lavin, S., Alegre, F., Cuenca, R., 2008. Ingesta passage
and gastric emptying times in loggerhead sea turtles (Caretta caretta). Res. Vet. Sci.
84 (1), 132–139.
Wallace, B., DiMatteo, A., Hurley, B., Finkbeiner, E., Bolten, A., Chaloupka, M.,
Hutchinson, B., Abreu-Grobois, F., Amorocho, D., Bjorndal, K., Bourjea, J., Bowen, B.,
Dueñas, R., Casale, P., Choudhury, B., Costa, A., Dutton, P., Fallabrino, A., Girard, A.,
Girondot, M., Godfrey, M., Hamann, M., López-Mendilaharsu, M., Marcovaldi, M.,
Mortimer, J., Musick, J., Nel, R., Pilcher, N., Seminoff, J., Troëng, S., Witherington,
B., Mast, R., 2010. Regional management units for marine turtles: a novel framework
for prioritizing conservation and research across multiple scales. PLoS One 5 (12),
e15465. https://doi.org/10.1371/journal.pone.0015465.
Wallace, B.P., Kor, C.Y., Dimatteo, A.D., Lee, T., Crowder, L.B., Lewison, R.L., 2013.
Impacts of fisheries bycatch on marine turtle populations worldwide: toward con-
servation and research priorities. Ecosphere 4, 1–49. https://doi.org/10.1890/ES12-
00388.1.
Zardus, J., Hadfield, M., 2004. Larval development and complemental males in
Chelonibia Testudinaria, a barnacle commensal with sea turtles. J. Crustac. Biol. 24
(3), 409–421.