Toxicity and Starvation Induce Major Trophic Isotope Variation in Daphnia Individuals: A Diet Switch Experiment Using Eight Phytoplankton Species of Differing Nutritional Quality.

Stable isotope values can express resource usage by organisms, but their precise interpretation is predicated using a controlled experiment-based validation process. Here, we develop a stable isotope tracking approach towards exploring resource shifts in a key primary consumer species Daphnia magna. We used a diet switch experiment and model fitting to quantify the stable carbon (δ13C) and nitrogen (δ15N) isotope turnover rates and discrimination factors for eight dietary sources of the plankton species that differ in their cellular organization (unicellular or filamentous), pigment and nutrient compositions (sterols and polyunsaturated fatty acids), and secondary metabolite production rates. We also conduct a starvation experiment. We evaluate nine tissue turnover models using Akaike's information criterion and estimate the repetitive trophic discrimination factors. Using the parameter estimates, we calculate the hourly stable isotope turnover rates. We report an exceedingly faster turnover value following dietary switching (72 to 96 h) and a measurable variation in trophic discrimination factors. The results show that toxic stress and the dietary quantity and quality induce trophic isotope variation in Daphnia individuals. This study provides insight into the physiological processes that underpin stable isotope patterns. We explicitly test multiple alternative dietary sources and fasting and discuss the parameters that are fundamental for field- and laboratory-based stable isotope studies.

Dietary nutrient allocation to somatic tissue synthesis in emerging subimago freshwater mayfly Ephemera danica.

BACKGROUND: The relative importance of nutrients derived from different sources for tissue synthesis is crucial for predicting a species responds to changes in food availability. The ecological and physiological strategies that govern the incorporation and routing of nutrients for reproduction are often well understood. However, the role and adaptive value of both species and individual variation during early life-stage remain elusive. In freshwater systems, dietary nutrient allocation to somatic tissue should be favoured when dietary source peaks and resource limitation may hinder flexible resource allocation. We used carbon and nitrogen stable isotopes (δ13C and δ15N) to examine metabolic nutrient routing and resource allocation from four dietary sources used to biosynthesize three somatic tissues of emerging subimago Ephemera danica. Aquatic emerging insects, such as the mayfly E. danica, are well suited for such studies. This is because, while burrowing nymph phase is a detritivores feeders with several early life-stages of metamorphosis, adult insects do not feed during this period but do utilize energy. RESULTS: Constructed models to predict percent proportional contribution of source to tissue showed that terrestrial detritus was the dominant nutrient source for abdomen, head and wing with mean values of 57%, 65% and 73%, respectively. There was evidence for differential resource allocation, as insect partitioned periphyton and sediment (but also seston) elements for tissue synthesis. Utilizing individual-specimen based relationship in isotope value; we derived tissue specific isotopic niche estimates, for the different tissue-source combinations. CONCLUSIONS: Results indicate that tissue selection is crucial for isotopic ecological measurements in arthropods. Mayfly has long been used as bio-indicator of freshwater ecosystems and their larvae show rapid response to environmental changes. In light of the recent evidence of drastic reduction in flying insect mass in Germany, developing a system using isotopic tools to trace nutrient flow in this important taxon will assist conservation and management efforts.

A European Multi Lake Survey dataset of environmental variables, phytoplankton pigments and cyanotoxins.

Under ongoing climate change and increasing anthropogenic activity, which continuously challenge ecosystem resilience, an in-depth understanding of ecological processes is urgently needed. Lakes, as providers of numerous ecosystem services, face multiple stressors that threaten their functioning. Harmful cyanobacterial blooms are a persistent problem resulting from nutrient pollution and climate-change induced stressors, like poor transparency, increased water temperature and enhanced stratification. Consistency in data collection and analysis methods is necessary to achieve fully comparable datasets and for statistical validity, avoiding issues linked to disparate data sources. The European Multi Lake Survey (EMLS) in summer 2015 was an initiative among scientists from 27 countries to collect and analyse lake physical, chemical and biological variables in a fully standardized manner. This database includes in-situ lake variables along with nutrient, pigment and cyanotoxin data of 369 lakes in Europe, which were centrally analysed in dedicated laboratories. Publishing the EMLS methods and dataset might inspire similar initiatives to study across large geographic areas that will contribute to better understanding lake responses in a changing environment.

Seasonal bat activity related to insect emergence at three temperate lakes.

Knowledge of aquatic food resources entering terrestrial systems is important for food web studies and conservation planning. Bats, among other terrestrial consumers, often profit from aquatic insect emergence and their activity might be closely related to such events. However, there is a lack of studies which monitor bat activity simultaneously with aquatic insect emergence, especially from lakes. Thus, our aim was to understand the relationship between insect emergence and bat activity, and investigate whether there is a general spatial or seasonal pattern at lakeshores. We assessed whole-night bat activity using acoustic monitoring and caught emerging and aerial flying insects at three different lakes through three seasons. We predicted that insect availability and seasonality explain the variation in bat activity, independent of the lake size and characteristics. Spatial (between lakes) differences of bat activity were stronger than temporal (seasonal) differences. Bat activity did not always correlate to insect emergence, probably because other factors, such as habitat characteristics, or bats' energy requirements, play an important role as well. Aerial flying insects explained bat activity better than the emerged aquatic insects in the lake with lowest insect emergence. Bats were active throughout the night with some activity peaks, and the pattern of their activity also differed among lakes and seasons. Lakes are important habitats for bats, as they support diverse bat communities and activity throughout the night and the year when bats are active. Our study highlights that there are spatial and temporal differences in bat activity and its hourly nocturnal pattern, that should be considered when investigating aquatic-terrestrial interactions or designing conservation and monitoring plans.

Dual-tracer-based isotope turnover rates in a highly invasive mysid Limnomysis benedeni from Lake Constance.

Understanding the ecological patterns of invasive species and their habitats require an understanding of the species' foraging ecology. Stable carbon (δ13C) and nitrogen (δ15N) isotope values provide useful information into the study of animal ecology and evolution, since the isotope ratios of consumers reflect consumer's dietary patterns. Nevertheless, the lack of species- and element-specific laboratory-derived turnover rates could limit their application. Using a laboratory-based dual stable isotope tracer approach (Na15 NO 3 and NaH13 CO 3), we evaluated the δ15N and δ13C isotope turnover rates in full-grown adult invasive Limnomysis benedeni from Lake Constance. We provide δ15N and δ13C turnover rates based on nonlinear least-squares regression and posterior linear regression models. Model precisions and fit were evaluated using Akaike's information criterion. Within a couple of days, the δ15N and δ13C of mysids began to change. Nevertheless, after about 14 days, L. benedeni did not reach equilibrium with their new isotope values. Since the experiment was conducted on adult subjects, it is evident that turnover was mainly influenced by metabolism (in contrast to growth). Unlike traditional dietary shifts, our laboratory-based dual stable isotope tracer approach does not shift the experimental organisms into a new diet and avoids dietary effects on isotope values. Results confirm the application of isotopic tracers to label mysid subpopulations and could be used to reflect assimilation and turnover from the labeled dietary sources. Field-based stable isotope studies often use isotopic mixing models commonly assuming diet-tissue steady state. Unfortunately, in cases where the isotopic composition of the animal is not in equilibrium with its diet, this can lead to highly misleading conclusions. Thus, our laboratory-based isotopic incorporation rates assist interpretation of the isotopic values from the field and provide a foundation for future research into using isotopic tracers to investigate invasion ecology.

Long-term development of hypolimnetic oxygen depletion rates in the large Lake Constance.

This study investigates over 30 years of dissolved oxygen dynamics in the deep interior of Lake Constance (max. depth: 250 m). This lake supplies approximately four million people with drinking water and has undergone strong re-oligotrophication over the past decades. We calculated depth-specific annual oxygen depletion rates (ODRs) during the period of stratification and found that 50% of the observed variability in ODR was already explained by a simple separation into a sediment- and volume-related oxygen consumption. Adding a linear factor for water depth further improved the model indicating that oxygen depletion increased substantially along the depth. Two other factors turned out to significantly influence ODR: total phosphorus as a proxy for the lake's trophic state and mean oxygen concentration in the respective depth layer. Our analysis points to the importance of nutrient reductions as effective management measures to improve and protect the oxygen status of such large and deep lakes.

The Effect of Parasite Infection on Stable Isotope Turnover Rates of δ15N, δ13C and δ34S in Multiple Tissues of Eurasian Perch Perca fluviatilis.

Stable isotope analysis of commercially and ecologically important fish can improve understanding of life-history and trophic ecology. However, accurate interpretation of stable isotope values requires knowledge of tissue-specific isotopic turnover that will help to describe differences in the isotopic composition of tissues and diet. We performed a diet-switch experiment using captive-reared parasite-free Eurasian perch (Perca fluviatilis) and wild caught specimens of the same species, infected with the pike tapeworm Triaenophorus nodulosus living in host liver tissue. We hypothesize that metabolic processes related to infection status play a major role in isotopic turnover and examined the influence of parasite infection on isotopic turn-over rate of carbon (δ13C), nitrogen (δ15N) and sulphur (δ34S) in liver, blood and muscle. The δ15N and δ13C turnovers were fastest in liver tissues, followed by blood and muscle. In infected fish, liver and blood δ15N and δ13C turnover rates were similar. However, in infected fish, liver and blood δ13C turnover was faster than that of δ15N. Moreover, in infected subjects, liver δ15N and δ13C turnover rates were three to five times faster than in livers of uninfected subjects (isotopic half-life of ca.3-4 days compared to 16 and 10 days, respectively). Blood δ34S turnover rate were about twice faster in non-infected individuals implying that parasite infection could retard the turnover rate of δ34S and sulphur containing amino acids. Slower turnover rate of essential amino acid could probably decrease individual immune function. These indicate potential hidden costs of chronic and persistent infections that may have accumulated adverse effects and might eventually impair life-history fitness. For the first time, we were able to shift the isotope values of parasites encapsulated in the liver by changing the dietary source of the host. We also report variability in isotopic turnover rates between tissues, elements and between infected and parasite-free individuals. These results contribute to our understanding of data obtained from field and commercial hatcheries; and strongly improve the applicability of the stable isotope method in understanding life-history and trophic ecology of fish populations.

Induced responses to grazing by an insect herbivore (Acentria ephemerella) in an immature macrophyte (Myriophyllum spicatum): an isotopic study.

While the mechanisms by which adult terrestrial plants deploy constitutive and induced responses to grazing pressure are well known, the means by which young aquatic plants defend themselves from herbivory are little studied. This study addresses nitrogen transport in the aquatic angiosperm Myriophyllum spicatum in response to herbivore exposure. Nitrogen tracers were used to monitor nitrogen uptake and reallocation in young plants in response to grazing by the generalist insect herbivore Acentria ephemerella. Total nitrogen content (N%) and patterns of nitrogen uptake and allocation (δ(15)N) were assessed in various plant tissues after 24 and 48 h. Following 24 h exposure to herbivore damage (Experiment 1), nitrogen content of plant apices was significantly elevated. This rapid early reaction may be an adaptation allowing the grazer to be sated as fast as possible, or indicate the accumulation of nitrogenous defense chemicals. After 48 h (Experiment 2), plants' tips showed depletion in nitrogen levels of ca. 60‰ in stem sections vulnerable to grazing. In addition, nitrogen uptake by grazed and grazing-prone upper plant parts was reduced and nutrient allocation into the relatively secure lower parts increased. The results point to three conclusions: (1) exposure to an insect herbivore induces a similar response in immature M. spicatum as previously observed in mature terrestrial species, namely a rapid (within 48 h) reduction in the nutritional value (N%) of vulnerable tissues, (2) high grazing intensity (100% of growing tips affected) did not limit the ability of young plants to induce resistance; and (3) young plants exposed to herbivory exhibit different patterns of nutrient allocation in vulnerable and secure tissues. These results provide evidence of induced defense and resource reallocation in immature aquatic macrophytes which is in line with the responses shown for mature aquatic macrophytes and terrestrial plants.

Tracking diet preferences of bats using stable isotope and fatty acid signatures of faeces.

Stable isotope and fatty acid signatures of biomaterials can provide important information about the dietary niche of animals. Stable isotope and fatty acid signatures differ between aquatic and terrestrial food webs, and therefore can be used to assess the aquatic and terrestrial contributions to the diets of species. We studied faecal samples of three co-occurring bat species with known differences in feeding preferences. The aim was to assess whether stable isotope and fatty acid signatures of faeces can be used to determine feeding preferences. We used bat faeces because they can be easily and non-invasively collected. We hypothesised that faeces stable isotope and fatty acid signatures will reveal the terrestrial, aquatic and mixed feeding niches of Myotis myotis, M. daubentonii, and M. mystacinus, respectively. As predicted, the faeces of M. myotis were characterized by higher δ(13)C values and higher concentrations of linoleic acid and total ω6 polyunsaturated fatty acids (PUFAs), which are typically higher in terrestrial food webs. The faeces of M. daubentonii had higher δ(15)Ν values and higher concentrations of docosahexaenoic acid and total ω3 PUFAs, characteristic features of aquatic systems. Myotis mystacinus faeces had intermediate δ(15)Ν values and concentrations of both types of fatty acids. Our results show that analysing stable isotope and/or fatty acid signatures of faeces provides a promising, non-invasive tool to study the feeding ecology of bats and to assess aquatic-terrestrial interactions.

Absence of sterols constrains food quality of cyanobacteria for an invasive freshwater bivalve.

The accumulation of cyanobacterial biomass may severely affect the performance of aquatic consumers. Here, we investigated the role of sterols in determining the food quality of cyanobacteria for the invasive clam Corbicula fluminea, which has become a common benthic invertebrate in many freshwater ecosystems throughout the world. In standardized growth experiments, juvenile clams were fed mixtures of different cyanobacteria (Anabaena variabilis, Aphanothece clathrata, Synechococcus elongatus) or sterol-containing eukaryotic algae (Cryptomonas sp., Nannochloropsis limnetica, Scenedesmus obliquus). In addition, the cyanobacterial food was supplemented with different sterols. We provide evidence that somatic growth of C. fluminea on cyanobacterial diets is constrained by the absence of sterols, as indicated by a growth-enhancing effect of sterol supplementation. Thus, our findings contribute to our understanding of the consequences of cyanobacterial mass developments for benthic consumers and highlight the importance of considering sterols as potentially limiting nutrients in aquatic food webs.

Alteration of kairomone-induced antipredator response of the freshwater amphipod Gammarus roeseli by sediment type.

The importance of sediment type and of kairomones released by juvenile burbot (Lota Iota) on behavioral responses of the freshwater amphipod Gammarus roeseli was studied in laboratory experiments. In the absence of kairomones. G. roeseli preferred coarse substrates; the highest preference was shown for the most coarse sediment. In the presence of kairomones, the amphipods preferred the less-coarse substrate pebbles over gravel. This difference is an effective antipredator response; G. roeseli is able to hide efficiently in the interstices of the pebbles. Juvenile burbot have a low foraging efficiency on pebbles and preferentially forage in the interstices of coarser sediments. G. roeseli responded differently to kairmones in different sediment types. The amphipods decreased their activity only in pebbles. In gravel, the rate of exploration behavior increased, and on a bare surface, the area containing fish kairomones was avoided.

A model approach to the population dynamics of the rotifer Brachionus rubens in two-stage chemostat culture.

A model, based on energy-flow considerations, is presented which describes the population dynamics of Brachionus rubens in the second stage of a two-stage algalrotifer chemostat. The rotifers are foodlimited with substrate-inhibition occurring at high algal densities. The model shows two stable states: steady state with constant density of rotifers and washout of the animals. Which one of the stable states is reached depends on the initial conditions.Empirical data are in general agreement with the model. Deviations may be explained by the fact that the data underlying the model calculations are based on a different food alga (Chlorella vulgaris) than the one used in the experiments (Monoraphidium minutum).The observed population growth rate reaches a maximum value of 0.84 (1/day) at algal densities of 3-4. 106 cells/ml. It decreases at higher algal densities. The egg ratio is related linearily to algal density without being reduced at high algal densities.