Dietary-based developmental plasticity

royalsocietypublishing.org/journal/rspb
affects juvenile survival in an
aquatic detritivore
Moritz D. Lürig1,2,3 and Blake Matthews2
Research 1
Department of Biology, Lund University, 22362 Lund, Sweden
2
Cite this article: Lürig MD, Matthews B. 2021 Department of Fish Ecology and Evolution, and 3Department of Aquatic Ecology, Eawag, Seestrasse 79,
6047 Kastanienbaum, Switzerland
Dietary-based developmental plasticity affects
juvenile survival in an aquatic detritivore. MDL, 0000-0002-8175-6234

Proc. R. Soc. B 288: 20203136.

Developmental plasticity is ubiquitous in natural populations, but the
https://doi.org/10.1098/rspb.2020.3136 underlying causes and fitness consequences are poorly understood. For con-
Downloaded from https://royalsocietypublishing.org/ on 09 October 2023

sumers, nutritional variation of juvenile diets is probably associated with

plasticity in developmental rates, but little is known about how diet quality
can affect phenotypic trajectories in ways that might influence survival to
Received: 21 December 2020 maturity and lifetime reproductive output. Here, we tested how the diet
Accepted: 19 January 2021 quality of a freshwater detritivorous isopod (Asellus aquaticus), in terms of
elemental ratios of diet (i.e. carbon : nitrogen : phosphorus; C : N : P), can
affect (i) developmental rates of body size and pigmentation and (ii)
variation in juvenile survival. We reared 1047 individuals, in a full-sib
split-family design (29 families), on either a high- (low C : P, C : N) or low-
Subject Category: quality (high C : P, C : N) diet, and quantified developmental trajectories of
Ecology body size and pigmentation for every individual over 12 weeks. Our diet
contrast caused strong divergence in the developmental rates of pigmenta-
Subject Areas: tion but not growth, culminating in a distribution of adult pigmentation
ecology spanning the broad range of phenotypes observed both within and among
natural populations. Under low-quality diet, we found highest survival at
Keywords: intermediate growth and pigmentation rates. By contrast, survival under
high-quality diet survival increased continuously with pigmentation rate,
development, stoichiometry, fitness, life
with longest lifespans at intermediate growth rates and high pigmentation
history, phenotypic plasticity, diet quality rates. Building on previous work which suggests that visual predation med-
iates the evolution of cryptic pigmentation in A. aquaticus, our study shows
how diet quality and composition can generate substantial phenotypic vari-
Author for correspondence: ation by affecting rates of growth and pigmentation during development in
the absence of predation.
Moritz D. Lürig
e-mail: [email protected]

1. Introduction
Developmental plasticity, when the phenotypic expression of genotypes depends
on the environmental conditions during development, is ubiquitous in animals
[1–3]. There are several mechanisms by which environmental conditions can
affect the phenotypic trajectories of individual juveniles [4,5], and several ways
in which such developmental plasticity can affect fitness variation: for example,
juveniles can experience physiological trade-offs that manifest in lowered per-
formance, such as reduced locomotion [6,7] or maintenance of basic body
functions [8], that might ultimately increase mortality prior to adulthood [1,9].
Over an individual’s lifetime, the environmental dependence of phenotypic
expression can weaken (e.g. irreversible developmental plasticity), and, in
some cases, can culminate in adult phenotypes that are maladaptive. Cryptic
Electronic supplementary material is available coloration, for example, is often determined during early developmental
online at https://doi.org/10.6084/m9.figshare. environments in response to potentially imperfect environmental cues about
predation risk in adult environments [10,11]. Despite the ubiquity of develop-
c.5291646.
mental plasticity, surprisingly little is known about the ecological factors

© 2021 The Author(s) Published by the Royal Society. All rights reserved.
 affecting divergence in developmental trajectories and the isopod A. aquaticus, we manipulated stoichiometric ratios 2
consequences of these trajectories for fitness variation. and availability of pigmentation precursors (i.e. tryptophan)

royalsocietypublishing.org/journal/rspb
The dietary quality of resources throughout juvenile and tracked individual growth and pigmentation rates, as
development is probably an important cause of developmental well as survival, of over 1000 individuals from 29 families.
plasticity, because of its potentially large effects on the Specifically, our rearing experiment allowed us to investigate
expression of morphological, physiological and behavioural (i) the extent of developmental plasticity in growth and pig-
traits of adults [12,13]. Across their lifetimes, organisms need mentation caused by our diet manipulations, and (ii) how
to balance the allocation of acquired resources for growth, such variation in developmental rates of growth and pigmen-
maintenance and reproduction [1,2,14]. Especially during tation can jointly affect the survival of juveniles, in the absence
early life, when investments in somatic growth are high of predators or their cues [27,33]. Based on previous work
[15,16], developmental trajectories might be more susceptible regarding the physiological mechanisms of isopod develop-
to variation in both resource quantity and quality [17,18]. ment [27,36,38], we expected to find higher pigmentation
The stoichiometric composition of essential elements (carbon, rates under a high-quality (= high-protein) diet. Moreover,
nitrogen and phosphorus) varies broadly among primary we anticipated associations between developmental rates of

Proc. R. Soc. B 288: 20203136

producers within and across ecosystems [19], and is a useful growth and pigmentation, partly because high-quality diets
proxy of variation in diet quality of consumers [20]. Substantial often covary with pigmentation precursors—a covariation
mismatches between consumers and their diets are common that we attempted to disentangle with our manipulation of
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[21–23], and if they occur early in development, they might tryptophan. Our results confirm pronounced developmen-
be an important ecological cause of plasticity [6,10,24] and of tal plasticity in pigmentation, and, to a lesser degree, in
fitness variation [25]. growth rates, and underscore the need to consider diet- or
The effects of diet variation on developmental trajectories resource-based developmental plasticity as an important
are likely to have important fitness consequences for consumers source of phenotypic variation, which may affect fitness
in general [3,26], and for detritivores in particular [27]. Dietary- before reproduction or selection from predation later in life.
based developmental plasticity can vary from maladaptive to
adaptive depending on the specific ecological context [3,28].
For example, high-quality diets that are available during juven- 2. Material and methods
ile development may allow organisms to reduce predation risk
(e.g. by outgrowing vulnerable stages or sizes [6], maturing ear- (a) Asellus aquaticus
lier [29] or expressing adult phenotypes that increase mating The freshwater isopod A. aquaticus is common in benthic commu-
success [30]). For detritivores, who have adapted in various nities across Europe and parts of Asia [41]. The small crustaceans
ways to low-quality food throughout their lifetime [31], we (mature animals are 4–15 mm; figure 1) are found in many differ-
might expect nutrition to be an important source of individual ent microhabitats, like beds of Chara tomentosa, Phragmites australis
(reed) or bare sand [33,34,36], and are considered to play a signifi-
variation in both developmental trajectories and fitness in
cant role in freshwater food webs [33,36,41]. While A. aquaticus
natural populations [32]. However, few studies (either of detri-
can feed on fresh plant material, they often prefer substrates
tivores or other consumers) have quantified how the link colonized with microbiota (i.e. bacteria and fungi; figure 1d ) on
between fitness variation and developmental trajectories of leaf litter or decaying macrophytes [36,42–44]. Feeding on
individuals depends on the nutritional quality of diets. fungal and microbial biofilms may help alleviate stoichiometric
The detritivorous freshwater isopod Asellus aquaticus is a mismatches between A. aquaticus and their nutritionally poor
useful model to explore how dietary variation can affect phe- detritial diets [36,43]. Moreover, the amino acid tryptophan,
notypic variation throughout juvenile development. Previous which is essential for the main pigment in A. aquaticus, is known
work in Swedish lakes has shown habitat-specificity of adult to vary strongly across various detrital resources [40], but
isopod pigmentation and body size [33,34]. The matching of neither the effects of tryptophan or nutrition have been investi-
body pigmentation with habitat backgrounds has been pri- gated in the context of isopod life history and development.
Here, we manipulate both diet quality and tryptophan availability
marily interpreted in the context of the evolution of crypsis
to explore the link between variation in developmental trajectories
in response to visual predation [33–35]. However, A. aquaticus
and juvenile survival.
also exhibits diet-based plasticity in terms of both growth
rate [36] and accumulation rates of pigmentation through
development [27]. At birth, isopods completely lack pigmen- (b) Common garden experiment
tation and become increasingly pigmented as they grow [27]. (i) Contrasts and food preparation
The development of pigmentation of A. aquaticus is cumulat- Using a common garden experiment, we quantified the extent of
ive and irreversible through development [37], and may be variation in developmental rates of growth and pigmentation,
linked to environmental sources of tryptophan, an amino and their effects on survival in A. aquaticus in response to diet com-
acid that is a metabolic precursor for the pigment xanthom- position (stoichiometric quality and tryptophan availability). To
matin [38,39]. Tryptophan varies strongly among detrital do so, we exposed 1047 juvenile isopods from 29 families shortly
resources of A. aquaticus [40], but neither the effects of tryp- after their birth (1–3 days) to four different dietary contrasts: high
tophan nor the dietary quality of resources has been elemental ratios (C : P and C : N, hereafter low-quality (LQ) diet)
and low elemental ratios (hereafter high-quality diet (HQ)), as
investigated in the context of survival variation of A. aquaticus
well as each of these diet combinations crossed with a supplement
through development.
(or not) of tryptophan. We measured growth, pigmentation and
Here, building on our previous work [27], we perform a survival of each individual over the course of 12 weeks. For
large laboratory experiment to test how varying dietary each family, half of the juveniles were randomly assigned to
environments affect developmental trajectories of juveniles, either low or high diet quality (full-sib/split-family design). For
and investigate the joint effects of diet and divergent develop- the eight families with the highest number of offspring (50–60
mental trajectories for juvenile fitness. Using the freshwater juveniles), we crossed the diet quality treatment with a
 3
(a) (d)

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5 mm

(e)

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1000

(b)
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C : P ratio
500

(c)
0
0 10 20 30 40
C : N ratio
isopods black alder leaf LQ-diet –T
(animal) black alder leaf (+fungi) HQ-diet –T
Chara tomentosa LQ-diet +T
Myriophyl/um spicatum HQ-diet +T

Figure 1. Phenotypic variation in pigmentation in the freshwater isopod A. aquaticus can be determined by diet. (a) Random sample of isopods taken from beds of
Chara tomentosa in Lake Lucerne at Kastanienbaum (measured with a flatbed scanner, brightness adjusted to match images from camera stand; size scale is for a–c).
(b,c) Example of an isopod reared under (b) low-quality and (c) high-quality diet (both no tryptophan, photographed with a camera stand). The levels of adult isopod
pigmentation measured throughout the diet manipulation fall well within the range of isopod pigmentation found in nature (figure 2d ) [27]. (d ) Isopods feeding on
fungi that form on the surface of alder leaves in standing water. (e) Elemental composition of various natural food items that isopods encounter in Lake Lucerne, as well
as the artificial diets used in this experiment (LQ, low quality/high elemental ratio; HQ, high quality/low elemental ratio; −T, without tryptophan supplement; +T, with
tryptophan supplement). This panel also shows the elemental composition of isopods collected from Lake Lucerne (black diamond). Elemental ratios are scaled by the
molar mass of the respective elements. The data for the figure can be found in electronic supplementary material, table S1. (Online version in colour.)

supplemental tryptophan treatment: in these eight families, 40 isopods were randomly distributed across jars (50 ml, PE),
juveniles were randomly distributed among high- and low-qual- which contained filtered lake water and a pellet of either of the
ity treatments, and the remaining 10–20 individuals among the diet types. We placed the jars inside racks that were arranged
two treatments with tryptophan supplement. For the high-quality randomly inside a flow-through water trough to buffer against
diet, we used 80% dry yeast (Saccharomyces cerevisiae) and 20% fluctuations in temperature. The set-up was maintained at 20°C
potato starch that was autoclaved together with agar and filtered with a 16 : 8 h light dark cycle, and temperature was controlled
lake water into a paste that was dried and cut into pellets (dry every day. We took pictures of all live isopods from each block
weight 1.2 ± 0.1 g). The low-quality diet was prepared in the every three weeks. Using small pipettes (for isopods bigger
same way, but with 20% yeast and 80% starch. For the tryptophan than approx. 5 mm, we used soft steel forceps), we transferred
supplement, we added 0.1 g of tryptophan per 1 g of food sub- an individual from its tube into a small container with lake
strate. We constructed these diets so as to capture some of the water, and from there onto a flat tray containing lake water
broad range of stoichiometric variation that isopods encounter underneath a camera mounted on a camera stand. After taking
in nature, from high-quality macrophyte detritus to low-quality the picture, we transferred each isopod into a new (autoclaved)
terrestrial detritus (figure 1). Our tryptophan manipulation unin- tube with fresh lake water and a new food pellet. We repeated
tentionally lowered the C : P of this diet treatment (figure 1e), but this procedure with every individual, yielding up to five
this effect was small relative to the overall diet contrast. phenotypic measurements for each developmental trajectory.

(ii) Experimental set-up and procedure (iii) Isopod pictures and phenotyping
We used juvenile isopods from a total of 29 successful matings We took pictures of isopods using a camera stand with a digital
(for details on isopod collection and breeding, see electronic single lens reflex camera (Canon) and a 100 mm macro lens
supplementary material) and started the common garden exper- (Tamron). The tray was uniformly illuminated with an LED spot
iment in three temporal blocks. From each family, juvenile ring (Leica). We ensured that each isopod specimen was flat on
 the tray, without movement or curling up. To quantify pigmenta- strongly affected by diet quality. Tryptophan only resulted 4
tion and body size of isopods from the digital images, we applied in significantly higher pigmentation rates under low-quality
computer vision techniques. For this purpose, we used the python

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diet (significant interaction diet quality × tryptophan; table 1
package phenopype [45]. It uses thresholding algorithms to seg- and figures 2 and 3). As indicated by the path analysis
ment isopods from the image background, to then extract the
(figure 3) and GAM2 (table 1 and figure 2), pigmentation
phenotypic information from the pixels marking the animal
rates were lowest when juveniles were reared under low-
(dorsal region of isopod torso = carapace, excluding legs and
quality diet and in the absence of the tryptophan supplement.
antennae). The greyscale values from these pixels were averaged
and converted to a pigmentation scale from 0 (greyscale value On the other hand, the tryptophan supplement resulted in
of 255) to 1 (greyscale value of 0). Body size was measured as car- slightly higher pigmentation rates under low-quality diet,
apace length, excluding legs and antennae. Previous work has but not under high-quality diet. This was indicated by a sig-
confirmed that phenopype results are highly correlated with nificant interactive effect of diet and tryptophan in GAM2
measurements of the same images using ImageJ (linear correlation (table 1 and figure 2) and in the path analysis (figure 3). Over-
between methods: slope = 0.98, R 2 = 0.97 [27]). all, and despite the presence of significant variation at the
family level for growth and pigmentation rates (see random

Proc. R. Soc. B 288: 20203136

effect of family in table 1; electronic supplementary material,
(c) Statistical analyses figure S2), the diet contrast resulted in clear divergence in
(i) Common garden experiment the build-up of pigmentation through development
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We tested for effects of diet composition and tryptophan supple- (figure 2b). For a given body size, these diet-induced differ-
ment on developmental rates of body size and pigmentation, as ences in pigmentation are comparable in magnitude to the
well as survival over the course of the experiment using a series
observed habitat-specific variation in nature (figure 2d ).
of generalized additive mixed models (GAM), using the ‘gamm’
Multiple lines of analysis indicate that there were complex
function in mgcv [46]. We fitted separate models each for body
size (GAM1; table 1) and pigmentation (log transformed,
interactions between diet quality and developmental rates
GAM2), with time separated by diet contrast as the fixed effect that affected survival of isopods. We found that survival of
and a thin plate spline term with time in weeks. Furthermore, juvenile isopods during the experiment depended strongly on
we fit a GAM with a binomial distribution family to test for both diet and tryptophan supplement: survival was much
differences in survival as a binary dependent variable, and higher on low-quality diets, and further increased by the
fixed effect and spline terms identical to the developmental tryptophan supplement. However, under a high-quality diet,
rate models (GAM3; table 1). All three models contained the tryptophan supplement did not affect survival (GAM3;
nested random terms for family and individual, and used diet table 1 and figure 3). Using the path analysis, we found that
as a parametric component in the spline terms. higher concurrent rates of growth and pigmentation also had
In a further step, we tested for effects of diet composition
a negative impact on survival independent of diet, as indicated
and of juvenile phenotypes right after birth on growth and pig-
by the interaction term (figure 3d). For a more in-depth analysis
mentation rates and survival by performing a path analysis
using Bayesian multilevel modelling [47]. In a single model, we
of the full three-way interaction of diet, growth rate and pig-
implemented three hierarchical levels, and included family as mentation rate, we used a multivariate additive framework,
the grouping term, allowing us to estimate relative effect sizes where we tested diet-specific relationships between both devel-
of developmental rates and starting conditions on lifespan opmental rates (GAM4; figure 4 and table 1). This analysis
under all diet treatment contrasts (see electronic supplementary revealed two distinct ‘survival surfaces’: under low-quality
material, table S2 for details). We applied both types of analysis diet, a single, high survival peak existed at intermediate
in a complementary fashion: with separate additive models, we growth and pigmentation rates. Survival under high quality
accounted for the nonlinearity in developmental rates, and was overall lower and varied nonlinearly across a wide range
with the path analysis, we were able to disentangle complex of both developmental rates (figure 4), as indicated by a signifi-
interactions linking rearing conditions and juvenile traits through
cant nonlinear interaction of diet and rates (table 1). Specifically,
development with survival variation.
survival on high-quality diet peaked at intermediate growth
To test for interactions between growth and pigmentation on
survival, we also applied a more complex multivariate GAM. To
rates and high rates of pigmentation (figure 4).
do so, we first converted measurements of body size and pigmen-
tation up until week 6 (dashed line in figure 2) to a single linear
slope per individual isopod (hereafter growth and pigmentation
rate, respectively). We chose to calculate slopes from this time 4. Discussion
frame, because pigmentation and growth increased linearly to Our experiment confirms and expands the results of a pre-
this point, and isopod survival up to this point was high. We vious study [27] that found diet-based developmental
then implemented an additive model (GAM4) with the ‘gam’ plasticity in pigmentation, and weak diet-based plasticity in
function from mgcv, using lifespan (in weeks) as the dependent growth in A. aquaticus. In the current paper, we found that
variable, single thin plate spline terms for growth and pigmenta-
growth of juvenile isopods was only weakly affected by our
tion rate, and a tensor smooth product term to test for the
manipulation of diet stoichiometry and the tryptophan sup-
interaction (table 1). The model included family as a random
effect, and the spline and tensor term included diet as a parametric
plement (figures 2a and 3a and table 1). The growth rates
component (see electronic supplementary material, for details). we measured are comparable to previous rearing exper-
iments that used naturally occurring food items [36],
confirming that the caloric content and nutritional balance
of the pellets that we provided ad libitum were an appropri-
3. Results ate rearing environment. Maintaining high growth rates on
We found that growth rates were only weakly affected by low-quality food might be an important mechanism in natu-
diet quality and tryptophan supplement (GAM 1; table 1 ral habitats to escape (outgrow) gape limited predators (e.g.
and figures 2 and 3), whereas rates of pigmentation were juvenile fish) or have a higher chance of escaping slow
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Table 1. Statistical results of generalized additive models. Models GAM1–GAM3 tested for an effect of diet quality content on growth, pigmentation and survival (figure 2), GAM4 tested for interactive effects of diet quality, growth
rates, pigmentation rates on survival of isopods (figure 4). Reported are results for linear (fixed effect) and nonlinear (smooth term) part of the model (tprs, thin plate regression spline; tp, tensor product). For each model, the degrees
of freedom for the fixed effect term are 1, and the number of knots for each smooth function is 3. Significance of random effects was tested with a likelihood ratio test. Significant (less than 0.05) and marginally significant (less than
0.1) results are in bold.

response smooth random

model variable fixed effect F p- value smooth term function edf F p-value effect d.f. Chisq p-value

GAM1 log(Length) diet 4.644 0.031 high-quality – T Tprs 2 4739.25 >0.001 individual 1 89.921 >0.001
tryptophan 3.434 0.064 high-quality +T tprs 1.99 603.079 >0.001 family 1 495.419 >0.001
diet × tryptophan 2.202 0.138 low-quality – T tprs 2 7036.52 >0.001 block 1 199.2 >0.001
low-quality +T tprs 2 1196.43 >0.001
GAM2 log(Pigmentation) diet 221.96 >0.001 high-quality – T tprs 1.96 1426.96 >0.001 individual 1 61.161 >0.001
tryptophan 2.735 0.098 high-quality +T tprs 1 271.881 >0.001 family 1 541.715 >0.001
diet × tryptophan 7.003 0.008 low-quality –T tprs 1.87 1179.35 >0.001 block 1 111.844 >0.001
low-quality +T tprs 1.9 267.761 >0.001
GAM3 survival diet 37.109 >0.001 high-quality T tprs 1.97 342.591 >0.001 individual 1 3318.86 >0.001
tryptophan 2.721 0.099 high-quality +T tprs 1.51 51.396 >0.001 family 1 384.212 >0.001
diet × tryptophan 7.71 0.006 low-quality −T tprs 1.95 324.69 >0.001 block 1 644.953 >0.001
low-quality +T tprs 1 58.669 >0.001
GAM4 survival diet 107.56 <0.001 high-quality × growth rate tprs 1.96 14.856 >0.001 family 1 23.466 0.217
growth rate 652.88 <0.002 low-quality × growth rate tprs 1.94 4.39 0.014 block 1 60.419 >0.001
pigmentation rate 246.89 <0.003 high-quality × pigmentation rate tprs 1 23.212 >0.001
diet × growth rate 108.07 <0.004 low-quality × pigmentation rate tprs 1.98 6.501 0.002
diet × pigmentation rate 66.537 <0.005 high-quality × growth rate × tp 3.21 7.755 >0.001
pigmentation rate
growth rate × pigmentation 2.709 0.1 low-quality+growth rate × tp 1 1.187 0.276
rate pigmentation rate
diet × growth rate × 2.778 0.09
pigmentation rate
5

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 (a) (b) 6
2.0

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−1.0
1.5
log(length (mm))

log(pigmentation)
−1.5
1.0

0.5 −2.0

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0 −2.5

0 3 6 9 12 0 3 6 9 12
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week week
(c) (d)
1.00 0.8

0.75
0.6
pigmentation
survival

0.50

0.4
0.25
Chara (CH)
Chara (SE)
reed (SE)
0 0.2
0 3 6 9 12 2 4 6 8
week length (mm)
low-quality −T low-quality +T
high-quality −T high-quality +T
Figure 2. Treatment-level model estimates (symbols) and family-level developmental trajectories (lines). The symbols with error bars show model estimates for log-
transformed length (a, GAM1), log-transformed pigmentation (b, GAM2) and survival (c, GAM3) for both diet contrasts (diet quality = circles, tryptophan = tri-
angles) at a given time point (details on the model statistics are given in table 1). Each line shows the family-level average of body size, pigmentation or
survival at a given time point. Solid lines indicate only protein manipulation; dashed lines indicate averages for the part of the families that were reared
under tryptophan supplement. The vertical line in a and b indicates the cut-off of values used for the multivariate additive model (t1–t3, GAM4). (d ) The untrans-
formed treatment-level averages for length and pigmentation at each time point (same symbol and colour coding as in a–c), and length and pigmentation of wild
caught isopods from different habitats. Differences in length and pigmentation due to the diet manipulation at the end of this experiment resemble phenotypic
variation in isopods from two different habitats in southern Sweden (SE, reed, black points; Chara tomentosa, dark grey points). Moreover, developmental trajectories
we measured in this experiment fall within the range of phenotypes of isopods collected from Lake Lucerne in Switzerland (CH, Chara tomentosa, light grey points).
(Online version in colour.)

moving invertebrate predators (e.g. odonate larvae) when rates of pigmentation, and also higher final levels pigmenta-
they are larger [35]. Although our diet contrast spanned tion at the end of the experiment. This is in agreement with a
beyond the range of natural food items that we measured previous study [27] and provides additional support for plas-
in our study population (figure 1), our treatments with ticity of pigmentation during juvenile development, which
high stoichiometric mismatch (i.e. high C : P/C : N) was is irreversible for adult isopods [33]. Indeed, our dietary
sufficient near natural growth [36] and pigmentation rates [27]. manipulations recapitulated the entire phenotypic range of
Pigmentation rates were strongly affected by our manipu- pigmentation for a given body size in the Lake Lucerne popu-
lation of diet stoichiometry (figures 2b and 3b and table 1): lation (see figures 1a–c and 2d) [27]. While variation among
when reared under high-quality diet (low C : P, C : N) juvenile families in the extent of phenotypic divergence probably
isopods from a majority of families (22 out of 29, electronic results from a mixture of genetic and environmental factors,
supplementary material, figure S2) showed greatly increased our experimental design can neither quantify additive genetic
 (c) 7
low-quality diet 0.38
(a)

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0.16
low-quality diet + (b)
0.27 growth rate
tryptophan
0.15 0.27
survival
tryptophan 0.17 pigmentation rate 0.25
during experiment
0.41
growth rate × (d)
juvenile length 0.32
pigmentation rate

juvenile pigmentation

(a) (b) (c) (d)

0.4 0.4 0.4

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1.0
pigmentation rate
growth rate

0.2 0.2 0.2 0.5

survival

survival
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0
0 0 0
–0.5

LQ HQ LQ HQ LQ HQ –2 –1 0 1
growth rate
LQ–T LQ+T
pigmentation rate 0.5 0 –0.5
HQ–T HQ+T

Figure 3. Path analysis using Bayesian multilevel modelling to investigate the effects of diet quality and tryptophan manipulation. Significant effects are indicated
by coloured arrows (green, positive; red, negative; grey, not significant (overlap of the posterior with zero)), effect sizes are given by number on arrows. Panels
illustrate the effects of the factorial manipulation of elemental composition (diet quality) and tryptophan on growth, pigmentation and survival rates (a–c, respect-
ively), as well as an interactive effect of growth and pigmentation rates on survival across all diet manipulations (d—full three-way interaction between diet, growth
and pigmentation rates are analysed by GAM 4 and shown in figure 4). Details on the path analysis are given in electronic supplementary material (table S2).
(Online version in colour.)

low-quality high-quality
0.015
12
predicted survival (weeks)

9
pigmentation rate (units d–1)

0.010
0 0.005 0.010
pigmentation rate (units d–1)

0.005

0 0.025 0.050 0.075 0 0.025 0.050 0.075

growth rate (mm d–1)

survival during low-quality –tryptophan

experiment (weeks) 3 6 9 12 high-quality +tryptophan

Figure 4. Survival landscapes modelled from the interaction of diet quality, growth rate and pigmentation rate (GAM4). Each point denotes an individual isopod
(black, quality contrast; grey, tryptophan contrast). Diet-specific surfaces are model estimates from GAM4 with survival during experiment as the dependent and diet-
specific growth and pigmentation rates between start and week 6 as the independent variable (see table 1 for details, GAM4). The blue (low-protein) and orange
lines (high-protein) show the predicted survival for a fixed growth rate of 0.05 mm d−1 over a range of pigmentation rates: under low-protein diet, a peak for high
survival is forming at intermediate growth and pigmentation rates, whereas under high-protein diet, survival increases linearly with pigmentation rate. (Online
version in colour.)
 variance of plasticity nor test for transgenerational plasticity Our experiment provided evidence for nonlinear inter- 8
(e.g. paternal effects). Even so, the high reproducibility of actions between diet quality and developmental rates that

royalsocietypublishing.org/journal/rspb
phenotypic divergence within families exposed to contrasting strongly affected juvenile survival. Specifically, under a low-
diets provides strong evidence for diet-based developmental quality diet (high C : P, C : N), survival was constrained
plasticity in our study population. around a single peak centred at intermediate growth and
Our supplement of tryptophan to both high- and low- pigmentation rates. By comparison, under a high-quality diet
quality diets showed small, but significant positive effects on (low C : P, C : N), high survival was observed over a broader
pigmentation rates, but only for isopods reared on low-quality range of growth and pigmentation rates, albeit with a
diet (figure 3b and table 1). It is well known that the addition tendency for high survival at intermediate growth rates.
of tryptophan to diets can increase pigmentation in insects. Previous work on other organisms has also observed broader
For example, larvae of cabbage butterflies (Pieris brassicae) survival landscapes on high versus low-quality food
reared on tryptophan-limited artificial foods have reduced [13,16,53]. However, this was not the case in our study (figure 4
wing pigmentation compared to larvae reared on tryptophan- inset): high-fitness under low-quality diet was constrained to a
rich foods [48]. Typically, organisms acquire tryptophan from single peak of moderate growth and pigmentation rates,

Proc. R. Soc. B 288: 20203136

protein-rich diets [49], and the yeast we used to create the whereas high-quality diet did not show a distinct high-fitness
high-quality diet (i.e. S. cerevisiae) is known to contain trypto- peak. This could either be due to the aforementioned negative
phan [50]. Therefore, the faster development of pigmentation consequences of protein breakdown, or to physiological stress
Downloaded from https://royalsocietypublishing.org/ on 09 October 2023

we observed in the low C : P diet could be partly explained by from accelerated rates of development [13,58].
higher levels of tryptophan originating from yeast. Previous work on populations of A. aquaticus in southern
A general result from our experiment was that juvenile Sweden has proposed that visual predation by predators is an
survival depended strongly on the developmental rates of important agent of selection, driving rapid evolution of cryptic
both growth and pigmentation, albeit in complex ways. body coloration in A. aquaticus [33,34]. Specifically, in shallow
Both the significant interaction in the path analysis (figure 3d) lakes, visual predators are thought to cause the evolution
and the multivariate additive model (figure 4) suggest that of darker isopods in dark stands of reed, and lighter isopods
fast-growing individuals had a lower likelihood of survival in light beds, of Chara tomentosa. However, the phenotypic
when they also had high rates of pigmentation (figure 3d). differences stemming from our diet manipulation caused
Previous work has suggested that elevated growth rates pigmentation differences as large as the phenotypic differen-
in A. aquaticus are associated with higher energy expendi- tiation observed in southern Sweden populations (figure 2d),
ture, and consequently, higher metabolism and resource but in the absence of predators or background variation.
requirements [51], which may explain why fast-growing indi- Additionally, we observe substantial variation in the slope and
viduals have higher mortality rates. Elevated dietary protein intercept of family-level reaction norms (electronic supplemen-
content has also been shown to reduce survival in other study tary material, figure S2) and a negative relationship between
systems [52,53], which is thought to be caused by energetic developmental trajectories and survival (figures 3 and 4 and
expenditure associated with protein digestion and potentially table 1). This suggests an important link between factors affect-
harmful breakdown products [37,49]. Moreover, it is possible ing development, and the phenotypic evolution of cryptic body
that a specific composition of the gut microbial community is coloration. In the light of this work, we need more direct tests of
required to digest certain proteins [54]. Still, only surprisingly the putative agents of selection driving phenotypic evolution
little is known about the direct effects of protein consumption and their mechanisms (e.g. macrophytes as diet and shelter).
for aquatic isopods and particularly A. aquaticus, given that The fact that we found elevated pigmentation rates under
many detrital food items may contain high amounts of low elemental ratios and tryptophan supplement adds com-
protein (figure 1). plexity to our understanding about how visual predators
Decreased survival under high developmental rates may might mediate the evolution of pigmentation in A. aquaticus
also be due to resource competition antagonisms within the (figure 2d). Certain macrophytes contain tryptophan in rela-
developing organism [15], namely if isopods experience tively high levels [40], but the breakdown of proteins
physiological costs of maintaining high rates of both containing tryptophan and their digestions may result in tox-
growth and pigmentation [13,17,18]. The relative consistency icity [37,49]. Ommochrome synthesis may be a mechanism
of growth rates across all treatment combinations suggests to bind excess tryptophan to pigment granules, while isopods
that the development of body size is more conserved than can take advantage of any high-quality biomass instead of
pigmentation [27]. Indeed, somatic growth, the correlated feeding selectively. Such ‘local excretion’ (i.e. the formation
development of thoracic and other tissues during early onto- of inert pigments from soluble tryptophan) might be adaptive
geny and before reaching maximum body size, is one main in arthropods to avoid toxicity of high-protein/low-elemental-
dimension of resource allocation in animals, followed by ratio diets [37]. Although not a direct test, our path analysis
physiological maintenance and reproduction [1,9,55]. How- provides some support for this hypothesis, as it shows
ever, depending on the resources available during early higher survival under high pigmentation rates and lower
ontogeny, development of secondary characteristics like growth rates (figure 3d). Such mechanisms do not exclude
ornaments, weapons or pigmentation can vary in compari- the possibility for the evolution of cryptic pigmentation, but
son to body size, due to the necessity to develop fully we need a better understanding of sources of tryptophan
sized body parts and organs to ensure their functionality in natural diets, and the associated costs of acquiring and
[56,57]. It is possible that during early ontogeny of A. aqua- using tryptophan to synthesize xanthommatin. Parasites,
ticus, resource allocation to growth is prioritized over the although known to affect pigmentation in A. aquaticus [39],
development of isopod pigmentation when stoichiometric unlikely played a role in our study because the isopods were
mismatches between consumers and their diet are high reared in filtered lake water and the diets were autoclaved
[15,19,25]. during their preparation.
 In our study, we explored the links between variation in could improve our understanding about the underlying 9
stoichiometric composition of diet, plasticity of developmental sources and fitness consequences of developmental plasticity

royalsocietypublishing.org/journal/rspb
rates and fitness of juveniles (figures 1, 2d and 4). Diet in natural populations.
stoichiometry and its potential mismatch with organisms’
nutritional requirements is increasingly acknowledged to Data accessibility. All relevant data, code and instructions for reproduc-
play a fundamental role in shaping life history and develop- tion of all shown results are available from the Dryad Digital
ment [12,24,59]. Our study illustrates the environmental Repository: https://doi.org/10.5061/dryad.1vhhmgqrt [63].
dependence of links between developmental rates and fitness Authors’ contributions. M.D.L. and B.M. conceived the idea and designed
variation in a natural population of detritivores. Such exper- the experiment, M.D.L. conducted all experimental and analytical
work. Both authors contributed equally to writing the manuscript.
iments, particularly if they are designed to test elemental
Competing interests. The authors declare that no conflicts of interest
stoichiometry and nutritional geometry theory [6,12], could affected the work on this paper.
be particularly insightful for consumers, including detritivores Funding. M.D.L. was funded by the Center for Adaptation to a Chan-
[60–62], that are likely to encounter stoichiometric mismatches ging Environment (ACE) at ETH Zürich, and by the Department of
through development [21–23]. Ultimately, such approaches Aquatic Ecology at Eawag.

Proc. R. Soc. B 288: 20203136

References
Downloaded from https://royalsocietypublishing.org/ on 09 October 2023

1. Stearns SC. 1992 The evolution of life histories. color preference for effective crypsis. Evol. Lett. 3, stoichiometry and nutritional geometry with
Oxford, UK: Oxford University Press. 313–320. (doi:10.1002/evl3.113) homeostasis concepts and integrative models of
2. West-Eberhard MJ. 2005 Developmental plasticity 11. Edelaar P, Jovani R, Gomez-Mestre I. 2017 Should I organism nutrition. Funct. Ecol. 31, 286–296.
and the origin of species differences. Proc. Natl change or should I go? Phenotypic plasticity and (doi:10.1111/1365-2435.12707)
Acad. Sci. USA 102(Suppl. 1), 6543–6549. (doi:10. matching habitat choice in the adaptation to 21. Martinson HM, Schneider K, Gilbert J, Hines JE,
1073/pnas.0501844102) environmental heterogeneity. Am. Nat. 190, Hambäck PA, Fagan WF. 2008 Detritivory:
3. Nettle D, Bateson M. 2015 Adaptive developmental 506–520. (doi:10.1086/693345) stoichiometry of a neglected trophic level. Ecol. Res.
plasticity: what is it, how can we recognize it and 12. Lee KP, Simpson SJ, Clissold FJ, Brooks R, Ballard 23, 487–491. (doi:10.1007/s11284-008-0471-7)
when can it evolve? Proc. R. Soc. B 282, 20151005. JWO, Taylor PW, Soran N, Raubenheimer D. 2008 22. Frainer A, Jabiol J, Gessner MO, Bruder A, Mckie BG.
(doi:10.1098/rspb.2015.1005) Lifespan and reproduction in Drosophila: new 2016 Stoichiometric imbalances between detritus
4. Fischer S, Bohn L, Oberhummer E, Nyman C, insights from nutritional geometry. Proc. Natl Acad. and detritivores are related to shifts in ecosystem
Taborsky B. 2017 Divergence of developmental Sci. USA 105, 2498–2503. (doi:10.1073/pnas. functioning. Oikos 125, 861–871. (doi:10.1111/oik.
trajectories is triggered interactively by early social 0710787105) 02687)
and ecological experience in a cooperative breeder. 13. Lee W-S, Monaghan P, Metcalfe NB. 2013 23. Halvorson HM, Sperfeld E, Evans-White MA. 2017
Proc. Natl Acad. Sci. USA 114, E9300–E9307. Experimental demonstration of the growth rate— Quantity and quality limit detritivore growth:
(doi:10.1073/pnas.1705934114) lifespan trade-off. Proc. R. Soc. B 280, 20122370. mechanisms revealed by ecological stoichiometry
5. Groothuis TGG, Taborsky B. 2015 Introducing (doi:10.1098/rspb.2012.2370) and co-limitation theory. Ecology 98, 2995–3002.
biological realism into the study of developmental 14. Naguib M, Podos J, Simmons LW, Barrett L, Healy (doi:10.1002/ecy.2026)
plasticity in behaviour. Front. Zool. 12(Suppl. 1), S6. SD, Zuk M (eds). 2017 Advances in the study of 24. Acharya K, Kyle M, Elser JJ. 2004 Biological
(doi:10.1186/1742-9994-12-S1-S6) behavior, vol. 49. Amsterdam, The Netherlands: stoichiometry of Daphnia growth: an
6. Lee W-S, Monaghan P, Metcalfe NB. 2010 The Elsevier Science. See https://www.sciencedirect.com/ ecophysiological test of the growth rate hypothesis.
trade-off between growth rate and locomotor bookseries/advances-in-the-study-of-behavior/vol/ Limnol. Oceanogr. 49, 656–665. (doi:10.4319/lo.
performance varies with perceived time until 49/suppl/C. 2004.49.3.0656)
breeding. J. Exp. Biol. 213, 3289–3298. (doi:10. 15. West-Eberhard MJ. 2003 Developmental plasticity 25. Leal MC, Seehausen O, Matthews B. 2017 The
1242/jeb.043083) and evolution. Oxford, UK: Oxford University Press. ecology and evolution of stoichiometric phenotypes.
7. Niewiarowski PH, Angilletta Jr MJ. 2008 See Https://market.android.com/details?id=book- Trends Ecol. Evol. 32, 108–117. (doi:10.1016/j.tree.
Countergradient variation in embryonic growth and 7DQNTPYaHlYC. 2016.11.006)
development: do embryonic and juvenile 16. Verberk WCEP, Siepel H, Esselink H. 2008 Life- 26. Klepsatel P, Knoblochová D, Girish TN, Dircksen H,
performances trade off? Funct. Ecol. 22, 895–901. history strategies in freshwater macroinvertebrates. Gáliková M. 2020 The influence of developmental
(doi:10.1111/j.1365-2435.2008.01441.x) Freshw. Biol. 53, 1722–1738. (doi:10.1111/j.1365- diet on reproduction and metabolism in Drosophila.
8. Chen E-H, Hou Q-L, Wei D-D, Jiang H-B, Wang J-J. 2427.2008.02035.x) BMC Evol. Biol. 20, 93. (doi:10.1186/s12862-020-
2017 Phenotypic plasticity, trade-offs and gene 17. Metcalfe NB, Monaghan P. 2001 Compensation 01663-y)
expression changes accompanying dietary restriction for a bad start: grow now, pay later? Trends 27. Lürig MD, Best RJ, Svitok M, Jokela J, Matthews B.
and switches in Bactrocera dorsalis (Hendel) Ecol. Evol. 16, 254–260. (doi:10.1016/S0169- 2019 The role of plasticity in the evolution of cryptic
(Diptera: Tephritidae). Sci. Rep. 7, 1988. (doi:10. 5347(01)02124-3) pigmentation in a freshwater isopod. J. Anim. Ecol.
1038/s41598-017-02106-3) 18. Metcalfe NB, Monaghan P. 2003 Growth versus 88, 612–623. (doi:10.1111/1365-2656.12950)
9. Reznick D. 2013 Evolution of life histories. In The lifespan: perspectives from evolutionary ecology. 28. Morris M, Rogers SM. 2013 Overcoming maladaptive
Princeton guide to evolution (eds JB Losos, DA Exp. Gerontol. 38, 935–940. (doi:10.1016/S0531- plasticity through plastic compensation. Curr. Zool.
Baum, DJ Futuyma, HE Hoekstra, RE 5565(03)00159-1) 59, 526–536. (doi:10.1093/czoolo/59.4.526)
Lenski, AJ Moore, CL Peichel, D Schluter, 19. Elser JJ et al. 2000 Nutritional constraints in 29. Plaistow SJ, Lapsley CT, Beckerman AP, Benton TG.
MC Whitlock), pp. 268–275. Princeton, NJ: terrestrial and freshwater food webs. Nature 408, 2004 Age and size at maturity: sex, environmental
Princeton University Press. 578–580. (doi:10.1038/35046058) variability and developmental thresholds.
10. van Bergen E, Beldade P. 2019 Seasonal plasticity in 20. Sperfeld E, Wagner ND, Halvorson HM, Malishev M, Proc. R. Soc. B 271, 919–924. (doi:10.1098/rspb.
anti-predatory strategies: matching of color and Raubenheimer D. 2017 Bridging ecological 2004.2682)
 30. Kodric-Brown A, Sibly RM, Brown JH. 2006 The 41. Sworobowicz L, Grabowski M, Mamos T, Burzyński 52. Fontana L, Partridge L. 2015 Promoting health and 10
allometry of ornaments and weapons. Proc. Natl A, Kilikowska A, Sell J, Wysocka A. 2015 Revisiting longevity through diet: from model organisms to

royalsocietypublishing.org/journal/rspb
Acad. Sci. USA 103, 8733–8738. (doi:10.1073/pnas. the phylogeography of Asellus aquaticus in Europe: humans. Cell 161, 106–118. (doi:10.1016/j.cell.
0602994103) insights into cryptic diversity and spatiotemporal 2015.02.020)
31. Nalepa CA, Bignell DE, Bandi C. 2001 Detritivory, diversification. Freshw. Biol. 60, 1824–1840. 53. Le Couteur DG, Solon-Biet S, Cogger VC, Mitchell SJ,
coprophagy, and the evolution of digestive (doi:10.1111/fwb.12613) Senior A, de Cabo R, Raubenheimer D, Simpson SJ.
mutualisms in Dictyoptera. Insectes Soc. 48, 42. Rossi L. 1985 Interactions between invertebrates 2016 The impact of low-protein high-carbohydrate
194–201. (doi:10.1007/PL00001767) and microfungi in freshwater ecosystems. Oikos 44, diets on aging and lifespan. Cell. Mol. Life Sci. 73,
32. Senior AM, Nakagawa S, Lihoreau M, Simpson SJ, 175–184. (doi:10.2307/3544059) 1237–1252. (doi:10.1007/s00018-015-2120-y)
Raubenheimer D. 2015 An overlooked consequence 43. Graça MA, Maltby L, Calow P. 1993 Importance of 54. Madsen L, Myrmel LS, Fjære E, Liaset B, Kristiansen
of dietary mixing: a varied diet reduces fungi in the diet of Gammarus pulex and Asellus K. 2017 Links between dietary protein sources, the
interindividual variance in fitness. Am. Nat. 186, aquaticus. I. Feeding strategies. Oecologia 93, gut microbiota, and obesity. Front. Physiol. 8, 1047.
649–659. (doi:10.1086/683182) 139–144. (doi:10.1007/BF00321203) (doi:10.3389/fphys.2017.01047)
33. Hargeby A, Johansson J, Ahnesjö J. 2004 Habitat- 44. Bohmann I. 2005 Coarse detritus in oligotrophic 55. Mousseau TA, Fox CW. 1998 Maternal effects as

Proc. R. Soc. B 288: 20203136

specific pigmentation in a freshwater isopod: lake littoral zones—utilization by invertebrates and adaptations. Oxford, UK: Oxford University Press.
adaptive evolution over a small spatiotemporal contribution to carbon flow. PhD thesis, University 56. Bonduriansky R, Day T. 2003 The evolution of static
scale. Evolution 58, 81–94. (doi:10.1111/j.0014- of Kalmar, Sweden. allometry in sexually selected traits. Evolution 57,
Downloaded from https://royalsocietypublishing.org/ on 09 October 2023

3820.2004.tb01575.x) 45. Lürig MD. 2018 phenopype—a phenotyping 2450–2458. (doi:10.1111/j.0014-3820.2003.

34. Hargeby A, Stoltz J, Johansson J. 2005 Locally pipeline for python. See https://doi.org/10.5281/ tb01490.x)
differentiated cryptic pigmentation in the freshwater zenodo.3483222. 57. Frankino WA, Zwaan BJ, Stern DL, Brakefield PM.
isopod Asellus aquaticus. J. Evol. Biol. 18, 713–721. 46. Wood SN. 2011 Fast stable restricted maximum 2005 Natural selection and developmental
(doi:10.1111/j.1420-9101.2004.00837.x) likelihood and marginal likelihood estimation of constraints in the evolution of allometries. Science
35. Eroukhmanoff F, Hargeby A, Arnberg NN, Hellgren O, semiparametric generalized linear models. J. R. Stat. 307, 718–720. (doi:10.1126/science.1105409)
Bensch S, Svensson EI. 2009 Parallelism and historical Soc. B 73, 3–36. (doi:10.1111/j.1467-9868.2010. 58. McCarthy ID, Houlihan Dominic F, Carter CG. 1994
contingency during rapid ecotype divergence in an 00749.x) Individual variation in protein turnover and growth
isopod. J. Evol. Biol. 22, 1098–1110. (doi:10.1111/j. 47. Bürkner P-C. 2018 Advanced Bayesian multilevel efficiency in rainbow trout, Oncorhynchus mykiss
1420-9101.2009.01723.x) modeling with the R package brms. R J. 10, (Walbaum). Proc. R. Soc. Lond. B 257, 141–147.
36. Marcus JH, Sutcliffe DW, Willoughby LG. 1978 395–411. (doi:10.32614/RJ-2018-017) (doi:10.1098/rspb.1994.0107)
Feeding and growth of Asellus aquaticus (Isopoda) 48. Kayser H. 1979 Ommochrome formation and 59. Elser JJ, O’brien WJ, Dobberfuhl DR, Dowling TE.
on food items from the littoral of Windermere, kynurenine excretion in Pieris brassicae: relation to 2000 The evolution of ecosystem processes: growth
including green leaves of Elodea canadensis. Freshw. tryptophan supply on an artificial diet. J. Insect rate and elemental stoichiometry of a key herbivore
Biol. 8, 505–519. (doi:10.1111/j.1365-2427.1978. Physiol. 25, 641–646. (doi:10.1016/0022- in temperate and arctic habitats. J. Evol. Biol. 13,
tb01473.x) 1910(79)90113-6) 845–853. (doi:10.1046/j.1420-9101.2000.00215.x)
37. Linzen B. 1974 The Tryptophan → Ommochrome 49. Arganda S, Bouchebti S, Bazazi S, Le Hesran S, Puga 60. Rietsma CS, Valiela I, Sylvester-Serianni A. 1982
pathway in insects. In Advances in insect physiology C, Latil G, Simpson SJ, Dussutour A. 2017 Parsing Food preferences of dominant salt marsh herbivores
(eds JE Treherne, MJ Berridge, VB Wigglesworth), the life-shortening effects of dietary protein: effects and detritivores. Mar. Ecol. 3, 179–189. (doi:10.
pp. 117–246. New York, NY: Academic Press. of individual amino acids. Proc. R. Soc. B 284, 1111/j.1439-0485.1982.tb00382.x)
38. Needham AE, Brunet PC. 1957 The integumental 20162052. (doi:10.1098/rspb.2016.2052) 61. Friberg N, Jacobsen D. 1994 Feeding plasticity of
pigment of Asellus. Cell. Mol. Life Sci. 13, 207–209. 50. Miozzari G, Niederberger P, Hütter R. 1978 two detritivore-shredders. Freshw. Biol. 32,
(doi:10.1007/BF02157167) Tryptophan biosynthesis in Saccharomyces cerevisiae: 133–142. (doi:10.1111/j.1365-2427.1994.tb00873.x)
39. Oetinger DF, Nickol BB. 1981 Effects of Acanthocephalans control of the flux through the pathway. J. Bacteriol. 62. Bloor MC. 2011 Dietary preference of Gammarus
on pigmentation of freshwater isopods. J. Parasitol. 67, 134, 48–59. (doi:10.1128/JB.134.1.48-59.1978) pulex and Asellus aquaticus during a laboratory
672–684. (doi:10.2307/3280441) 51. Peeters ETHM, Camu JM, Beijer JAJ, Scheffer M, breeding programme for ecotoxicological studies.
40. Muztar AJ, Slinger SJ, Burton JH. 1978 The chemical Gardeniers JJP. 2002 Response of the waterlouse Int. J. Zool. 2011, 1–5. (doi:10.1155/2011/294394)
composition of aquatic macrophytes. II. Amino acid Asellus aquaticus to multiple stressors: effects of 63. Lürig MD, Matthews B. 2021 Data from: Dietary-
composition of the protein and non-protein current velocity and mineral substratum. J. Aquat. based developmental plasticity affects juvenile
fractions. Can. J. Plant Sci. 58, 843–849. (doi:10. Ecosyst. Stress Recov. 9, 193–203. (doi:10.1023/ survival in an aquatic detritivore. Dryad Digital
4141/cjps78-123) A:1021218721123) Repository. (doi:10.5061/dryad.1vhhmgqrt)