Growth and fatty acid composition of pikeperch (Sander lucioperca L., 1758) larvae under altered feeding protocol including the copepod Apocyclops panamensis (Marsh, 1913).

Alternative live feeds for small and sensitive fish early life stages such as pikeperch (Sander lucioperca L., 1758) can improve the larval quantity, quality and performance in aquaculture. Therefore, this study evaluated the cyclopoid copepod Apocyclops panamensis (Marsh, 1913) as live feed for pikeperch larviculture from day 11 post hatch (dph) in two independent experiments. In both experiments, pikeperch larvae had the highest specific growth rate (SGR) when they fed on Brachionus plicatilis until dph 11 and A. panamensis until dph 16-18. SGR was related to a decrease in total fatty acids (FAs), saturated FAs and monounsaturated FAs in pikeperch larvae, indicating their use as energy for growth. Within the polyunsaturated FAs, docosahexaenoic acid (DHA) increased in larvae fed with A. panamensis and coincided with the highest SGR suggesting that DHA is accumulated in larvae as structural FA. Our study demonstrated a suitable pikeperch larval fatty acid composition for growth after feeding A. panamensis compared with Artemia sp. from dph 11 until dph 16 and previously fed with B. plicatilis. Moreover, it highlighted the importance of the dietary PUFAs in pikeperch rearing, specifically of linoleic acid (LA) from dph 4 until dph 11 and of DHA from dph 11 onwards.

Omega-3 versus Omega-6: Are We Underestimating the Ecological Significance of Arachidonic Acid in Aquatic Systems?

The long-chain polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (EPA, ω-3, or n-3) and arachidonic acid (ARA, ω-6 or n-6) are known to have distinct physiological functions, yet can both support growth and reproduction of consumers, raising the question of whether EPA and ARA are ecologically substitutable dietary resources. We explored the relative importance of EPA and ARA for the growth and reproduction of the freshwater keystone herbivore Daphnia in a life-history experiment. Both PUFA were supplemented in a concentration-dependent manner to a PUFA-free diet, separately and in combination (50% EPA: 50% ARA mixture). The growth-response curves obtained with EPA, ARA, and the mixture were virtually congruent and the thresholds for PUFA limitation did not differ, indicating that EPA (n-3) and ARA (n-6) were substitutable dietary resources under the applied experimental conditions. The actual requirements for EPA and ARA might change with growth conditions, e.g., under the influence of parasites or pathogens. The higher retention of ARA in Daphnia suggests that EPA and ARA are subject to different turnover rates, which also implies different physiological functions. Studies on the ARA requirements of Daphnia could provide valuable information on the presumably underestimated ecological importance of ARA in freshwater food webs.

Photoperiodic adaptation of aanat and clock gene expression in seasonal populations of Daphnia pulex.

Changes in day-length entrain the endogenous clock of organisms leading to complex responses to photoperiod. In long-lived organisms experiencing several seasons this response of the clock to photoperiod is phenotypically plastic. However, short-lived organisms often experience a single season without pronounced changes in day-length. For those, a plastic response of the clock to different seasons would not necessarily be adaptive. In aquatic ecosystems, zooplankton species like Daphnia live only for some weeks, i.e. one week up to ca. two months. However, they often show a succession of clones that are seasonally adapted to environmental changes. Here, we found that 16 Daphnia clones per each of three seasons ( = 48 clones) from the same pond and year differed in clock gene expression with a homogenous gene expression pattern in ephippia-hatched spring clones and a bimodal expression pattern in summer and autumn populations indicating an ongoing adaptation process. We clearly demonstrate that spring clones were adapted to a short, and summer clones to a long photoperiod. Furthermore, we found that gene expression of the melatonin-synthesis enzyme AANAT was always lowest in summer clones. In the Anthropocene, Daphnia's clock might be disturbed by light-pollution and global warming. Since Daphnia is a key-organism in trophic carbon transfer, a disruption of its clock rhythm would be devastating for the stability of freshwater ecosystems. Our results are an important step in understanding the adaptation of Daphnia's clock to environmental changes.

Outcomes and Risk Factors in Microsurgical Forefoot Reconstruction.

BACKGROUND: Defects at the forefoot frequently require microsurgical reconstruction; however, reconstructive failure can lead to results inferior to primary amputation. The purpose of this study was to identify independent factors affecting surgical outcomes and hospitalization time in these patients. METHODS: All patients that underwent free flap reconstruction of the forefoot between 2008 and 2019 were reviewed retrospectively. Statistical evaluation included binary logistic regression and correlation analysis. RESULTS: A total of 93 free flap procedures were performed in 87 patients. The most common defect etiologies were acute trauma (30 cases; 32.3%), diabetic foot syndrome (20 cases; 21.5%), and infection (17 cases; 18.3%). Muscle flaps were used in 50 cases (53.8%) and fasciocutaneous flaps were used in 43 cases (46.2%). Major complications occurred in 24 cases (25.8%) including 11 total flap losses and 2 partial flap losses. Minor complications occurred in 38 cases (40.9%). Patients aged 60 years or above were at significant higher risk of major complications (p = 0.029). Use of fasciocutaneous flaps (odds ratio [OR]: 14.341; p = 0.005), arterial hypertension (OR: 18.801; p = 0.014), and operative time (min) (OR: 1.010; p = 0.029) were identified as individual risk factors for major complications. Two venous anastomoses significantly reduced the risk of major complications (OR: 0.078; p = 0.022). Multiresistant bacterial wound colonization (OR: 65.152; p < 0.001) and defect size (OR: 1.007; p = 0.045) were identified as independent risk factors for minor complications. The median hospital stay was 28 days (7-85 days). Age significantly correlated with the length of hospital stay (r = 0.405, p < 0.01). CONCLUSION: Our study identified independent risk factors that might help to make individual decisions whether to target microsurgical forefoot reconstruction or primary amputation. Two venous anastomoses should be performed whenever feasible, and muscle free flaps should be preferred in patients at higher risk of major surgical complications.

Photosynthetic activity in both algae and cyanobacteria changes in response to cues of predation.

A plethora of adaptive responses to predation has been described in microscopic aquatic producers. Although the energetic costs of these responses are expected, with their consequences going far beyond an individual, their underlying molecular and metabolic mechanisms are not fully known. One, so far hardly considered, is if and how the photosynthetic efficiency of phytoplankton might change in response to the predation cues. Our main aim was to identify such responses in phytoplankton and to detect if they are taxon-specific. We exposed seven algae and seven cyanobacteria species to the chemical cues of an efficient consumer, Daphnia magna, which was fed either a green alga, Acutodesmus obliquus, or a cyanobacterium, Synechococcus elongatus (kairomone and alarm cues), or was not fed (kairomone alone). In most algal and cyanobacterial species studied, the quantum yield of photosystem II increased in response to predator fed cyanobacterium, whereas in most of these species the yield did not change in response to predator fed alga. Also, cyanobacteria tended not to respond to a non-feeding predator. The modal qualitative responses of the electron transport rate were similar to those of the quantum yield. To our best knowledge, the results presented here are the broadest scan of photosystem II responses in the predation context so far.

A sterol-mediated gleaner-opportunist trade-off underlies the evolution of grazer resistance to cyanobacteria.

The human-caused proliferation of cyanobacteria severely impacts consumers in freshwater ecosystems. Toxicity is often singled out as the sole trait to which consumers can adapt, even though cyanobacteria are not necessarily toxic and the lack of nutritionally critical sterols in cyanobacteria is known to impair consumers. We studied the relative significance of toxicity and dietary sterol deficiency in driving the evolution of grazer resistance to cyanobacteria in a large lake with a well-documented history of eutrophication and oligotrophication. Resurrecting decades-old Daphnia genotypes from the sediment allowed us to show that the evolution and subsequent loss of grazer resistance to cyanobacteria involved an adaptation to changes in both toxicity and dietary sterol availability. The adaptation of Daphnia to changes in cyanobacteria abundance revealed a sterol-mediated gleaner-opportunist trade-off. Genotypes from peak-eutrophic periods showed a higher affinity for dietary sterols at the cost of a lower maximum growth rate, whereas genotypes from more oligotrophic periods showed a lower affinity for dietary sterols in favour of a higher maximum growth rate. Our data corroborate the significance of sterols as limiting nutrients in aquatic food webs and highlight the applicability of the gleaner-opportunist trade-off for reconstructing eco-evolutionary processes.

Ecological impacts of photosynthetic light harvesting in changing aquatic environments: A systematic literature map.

Underwater light is spatially as well as temporally variable and directly affects phytoplankton growth and competition. Here we systematically (following the guidelines of PRISMA-EcoEvo) searched and screened the published literature resulting in 640 individual articles. We mapped the conducted research for the objectives of (1) phytoplankton fundamental responses to light, (2) effects of light on the competition between phytoplankton species, and (3) effects of climate-change-induced changes in the light availability in aquatic ecosystems. Among the fundamental responses of phytoplankton to light, the effects of light intensity (quantity, as measure of total photon or energy flux) were investigated in most identified studies. The effects of the light spectrum (quality) that via species-specific light absorbance result in direct consequences on species competition emerged more recently. Complexity in competition arises due to variability and fluctuations in light which effects are sparsely investigated on community level. Predictions regarding future climate change scenarios included changes in in stratification and mixing, lake and coastal ocean darkening, UV radiation, ice melting as well as light pollution which affect the underwater light-climate. Generalization of consequences is difficult due to a high variability, interactions of consequences as well as a lack in sustained timeseries and holistic approaches. Nevertheless, our systematic literature map, and the identified articles within, provide a comprehensive overview and shall guide prospective research.

Phenotypic Diversity and Plasticity of Photoresponse Across an Environmentally Contrasting Family of Phytoflagellates.

Organisms often employ ecophysiological strategies to exploit environmental conditions and ensure bio-energetic success. However, the many complexities involved in the differential expression and flexibility of these strategies are rarely fully understood. Therefore, for the first time, using a three-part cross-disciplinary laboratory experimental analysis, we investigated the diversity and plasticity of photoresponsive traits employed by one family of environmentally contrasting, ecologically important phytoflagellates. The results demonstrated an extensive inter-species phenotypic diversity of behavioural, physiological, and compositional photoresponse across the Chlamydomonadaceae, and a multifaceted intra-species phenotypic plasticity, involving a broad range of beneficial photoacclimation strategies, often attributable to environmental predisposition and phylogenetic differentiation. Deceptively diverse and sophisticated strong (population and individual cell) behavioural photoresponses were observed, with divergence from a general preference for low light (and flexibility) dictated by intra-familial differences in typical habitat (salinity and trophy) and phylogeny. Notably, contrasting lower, narrow, and flexible compared with higher, broad, and stable preferences were observed in freshwater vs. brackish and marine species. Complex diversity and plasticity in physiological and compositional photoresponses were also discovered. Metabolic characteristics (such as growth rates, respiratory costs and photosynthetic capacity, efficiency, compensation and saturation points) varied elaborately with species, typical habitat (often varying more in eutrophic species, such as Chlamydomonas reinhardtii), and culture irradiance (adjusting to optimise energy acquisition and suggesting some propensity for low light). Considerable variations in intracellular pigment and biochemical composition were also recorded. Photosynthetic and accessory pigments (such as chlorophyll a, xanthophyll-cycle components, chlorophyll a:b and chlorophyll a:carotenoid ratios, fatty acid content and saturation ratios) varied with phylogeny and typical habitat (to attune photosystem ratios in different trophic conditions and to optimise shade adaptation, photoprotection, and thylakoid architecture, particularly in freshwater environments), and changed with irradiance (as reaction and harvesting centres adjusted to modulate absorption and quantum yield). The complex, concomitant nature of the results also advocated an integrative approach in future investigations. Overall, these nuanced, diverse, and flexible photoresponsive traits will greatly contribute to the functional ecology of these organisms, addressing environmental heterogeneity and potentially shaping individual fitness, spatial and temporal distribution, prevalence, and ecosystem dynamics.

Field studies on breeding sites of Culicoides LATREILLE (Diptera: Ceratopogonidae) in agriculturally used and natural habitats.

Culicoides are vectors of pathogens mainly of veterinary importance. To establish targeted vector control measures, it is paramount to comprehend the ecological factors determining their distribution. Therefore, we used emergence traps to sample eight biotopes and assess their potential as breeding sites. Part one of the study investigates agricultural habitats, part two compares four biotopes of a forest-dominated area with less anthropogenic influence, including a physicochemical analysis of soil moisture, pH value and organic content. Thirteen culicoid species were collected, with a strong dominance of the Obsoletus Complex on meadows, and with Culicoides punctatus (MEIGEN), Culicoides pictipennis (STAEGER) and the Obsoletus Complex, to be the most abundant species in the natural habitats. Several co-existing species were found, some of them not having been described before. Our results suggest that ungrazed meadows seem unsuitable as breeding sites. Only the influence of livestock creates adequate conditions for certain midge species. The alder on fen site contained most culicoid species with the highest species diversity. Our study clearly indicates that knowledge of species-specific preferences for environmental habitat conditions (choice of breeding site) in connection to soil conditions is crucial to understand the biology and phenology of midges and their role as vectors of pathogens.

Geographic clines in Daphnia magna's circadian clock gene expression: Local adaptation to photoperiod.

Nearly all organisms show daily and seasonal physiological and behavioural responses that are necessary for their survival. Often these responses are controlled by the rhythmic activity of an endogenous clock that perceives day length. Day length differs not only between seasons but also along latitudes, with different seasonal day lengths between the north and the south. Both seasonal and latitudinal differences in day length are discussed to be perceived/processed by the endogenous clock. Some species are distributed over a wide range of latitudes; it should be highly adaptive for these species to be able to time physiological responses (e.g. migration behaviour and diapause) according to the organisms' respective photoperiod, i.e. their respective seasonal and latitudinal day length. The mediator of day length is the indoleamine hormone melatonin which is synthesized by melatonin-producing enzymes (AANAT and HIOMT). These enzymes are in turn controlled by an endogenous clock. The ubiquitous aquatic keystone organism Daphnia possess clock and melatonin synthesis genes that are rhythmically expressed over 24hours. We were able to show that the 24-h rhythm of D. magna's clock persists in constant darkness and is thus truly circadian. In one particular photoperiod, all D. magna clones produced a similar melatonin concentration due to a fixed AANAT activity. However, we have demonstrated that clones originating from different latitudes are adapted to their respective photoperiod by showing a geographic cline in clock and downstream melatonin synthesis gene expression. These findings hint at the problem locally adapted organisms face when they are forced to leave their respective photoperiod, e.g. because of climate change-driven range-expansion. If such a species is incapable of adjusting its endogenous clock to an unknown photoperiod, it will likely become extinct.

Inter- and intraspecific differences in rotifer fatty acid composition during acclimation to low-quality food.

Biochemical food quality constraints affect the performance of consumers and mediate trait variation among and within consumer species. To assess inter- and intraspecific differences in fatty acid retention and conversion in freshwater rotifers, we provided four strains of two closely related rotifer species, Brachionus calyciflorus sensu stricto and Brachionus fernandoi, with food algae differing in their fatty acid composition. The rotifers grazed for 5 days on either Nannochloropsis limnetica or Monoraphidium minutum, two food algae with distinct polyunsaturated fatty acid (PUFA) profiles, before the diets were switched to PUFA-free Synechococcus elongatus, which was provided for three more days. We found between- and within-species differences in rotifer fatty acid compositions on the respective food sources and, in particular, highly specific acclimation reactions to the PUFA-free diet. The different reactions indicate inter- but also intraspecific differences in physiological traits, such as PUFA retention, allocation and bioconversion capacities, within the genus Brachionus that are most likely accompanied by differences in their nutritional demands. Our data suggest that biochemical food quality constraints act differently on traits of closely related species and of strains of a particular species and thus might be involved in shaping ecological interactions and evolutionary processes. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Plant-soil feedback effects altered by aboveground herbivory explain plant species abundance in the landscape.

Relatively little is known about how plant-soil feedbacks (PSFs) may affect plant growth in field conditions where factors such as herbivory may be important. Using a potted experiment in a grassland, we measured PSFs with and without aboveground insect herbivory for 20 plant species. We then compared PSF values to plant landscape abundance. Aboveground herbivory had a large negative effect on PSF values. For 15 of 20 species, PSFs were more negative with herbivory than without. This occurred because plant biomass on "home" soils was smaller with herbivory than without. PSF values with herbivory were correlated with plant landscape abundance, whereas PSF values without herbivory were not. Shoot nitrogen concentrations suggested that plants create soils that increase nitrogen uptake, but that greater shoot nitrogen values increase herbivory and that the net effect of positive PSF and greater aboveground herbivory is less aboveground biomass. Results provided clear evidence that PSFs alone have limited power in explaining species abundances and that herbivory has stronger effects on plant biomass and growth on the landscape. Our results provide a potential explanation for observed differences between greenhouse and field PSF experiments and suggest that PSF experiments need to consider important biotic interactions, like aboveground herbivory, particularly when the goal of PSF research is to understand plant growth in field conditions.

Fitness response variation within and among consumer species can be co-mediated by food quantity and biochemical quality.

In natural heterogeneous environments, the fitness of animals is strongly influenced by the availability and composition of food. Food quantity and biochemical quality constraints may affect individual traits of consumers differently, mediating fitness response variation within and among species. Using a multifactorial experimental approach, we assessed population growth rate, fecundity, and survival of six strains of the two closely related freshwater rotifer species Brachionus calyciflorus sensu stricto and Brachionus fernandoi. Therefore, rotifers fed low and high concentrations of three algal species differing in their biochemical food quality. Additionally, we explored the potential of a single limiting biochemical nutrient to mediate variations in population growth response. Therefore, rotifers fed a sterol-free alga, which we supplemented with cholesterol-containing liposomes. Co-limitation by food quantity and biochemical food quality resulted in differences in population growth rates among strains, but not between species, although effects on fecundity and survival differed between species. The effect of cholesterol supplementation on population growth was strain-specific but not species-specific. We show that fitness response variations within and among species can be mediated by biochemical food quality. Dietary constraints thus may act as evolutionary drivers on physiological traits of consumers, which may have strong implications for various ecological interactions.

The relative importance of plant-soil feedbacks for plant-species performance increases with decreasing intensity of herbivory.

Under natural conditions, aboveground herbivory and plant-soil feedbacks (PSFs) are omnipresent interactions strongly affecting individual plant performance. While recent research revealed that aboveground insect herbivory generally impacts the outcome of PSFs, no study tested to what extent the intensity of herbivory affects the outcome. This, however, is essential to estimate the contribution of PSFs to plant performance under natural conditions in the field. Here, we tested PSF effects both with and without exposure to aboveground herbivory for four common grass species in nine grasslands that formed a gradient of aboveground invertebrate herbivory. Without aboveground herbivores, PSFs for each of the four grass species were similar in each of the nine grasslands-both in direction and in magnitude. In the presence of herbivores, however, the PSFs differed from those measured under herbivory exclusion, and depended on the intensity of herbivory. At low levels of herbivory, PSFs were similar in the presence and absence of herbivores, but differed at high herbivory levels. While PSFs without herbivores remained similar along the gradient of herbivory intensity, increasing herbivory intensity mostly resulted in neutral PSFs in the presence of herbivores. This suggests that the relative importance of PSFs for plant-species performance in grassland communities decreases with increasing intensity of herbivory. Hence, PSFs might be more important for plant performance in ecosystems with low herbivore pressure than in ecosystems with large impacts of insect herbivores.

Feeding in the frequency domain: coarser-grained environments increase consumer sensitivity to resource variability, covariance and phase.

Theory predicts that resource variability hinders consumer performance. How this effect depends on the temporal structure of resource fluctuations encountered by individuals remains poorly understood. Combining modelling and growth experiments with Daphnia magna, we decompose the complexity of resource fluctuations and test the effect of resource variance, supply peak timing (i.e. phase) and co-limiting resource covariance along a gradient from high to low frequencies reflecting fine- to coarse-grained environments. Our results show that resource storage can buffer growth at high frequencies, but yields a sensitivity of growth to resource peak timing at lower ones. When two resources covary, negative covariance causes stronger growth depression at low frequencies. However, negative covariance might be beneficial at intermediate frequencies, an effect that can be explained by digestive acclimation. Our study provides a mechanistic basis for understanding how alterations of the environmental grain size affect consumers experiencing variable nutritional quality in nature.

Temperature-induced changes in body lipid composition affect vulnerability to oxidative stress in Daphnia magna.

Optimizing physiological functions at different temperatures includes shifts in the lipid composition of ectothermic animals. These shifts may be associated with changes in lipid peroxidation in response to oxidative stress, because lipids differ in their susceptibility to oxidative damage. Polyunsaturated fatty acids (PUFA) are particular prone to peroxidation. Here, we analyzed changes in the fatty acid composition, cholesterol content and the level of oxidative damage as thiobarbituric reactive substances (TBARS) in Daphnia magna as a function of acclimation temperature. The total fatty acid content was highest in cold-acclimated animals. The relative share of most PUFA decreased with increasing acclimation temperature. In contrast, the contribution of saturated and monounsaturated fatty acids (SFA and MUFA) increased with acclimation temperature, although the latter to a lower extent. Cholesterol content remained unchanged. The level of oxidative damage was lowest in animals reared at warm temperatures, most likely reflecting their lowest content of PUFA. Heat exposure (1 h at 33 °C) caused the highest increase in lipid peroxidation in cold-acclimated animals, containing more PUFA. Our data suggest that cold-induced adjustments in the body lipid composition increase the vulnerability of zooplankton to heat-induced oxidative stress. In particular, animals performing diel vertical migration may be highly susceptible to temperature-induced lipid damage.

Nitrate or ammonium: Influences of nitrogen source on the physiology of a green alga.

In freshwaters, algal species are exposed to different inorganic nitrogen (Ni) sources whose incorporation varies in biochemical energy demand. We hypothesized that due to the lesser energy requirement of ammonium ( NH 4 + )-use, in contrast to nitrate ( NO 3 - )-use, more energy remains for other metabolic processes, especially under CO2- and phosphorus (Pi) limiting conditions. Therefore, we tested differences in cell characteristics of the green alga Chlamydomonas acidophila grown on NH 4 + or NO 3 - under covariation of CO2 and Pi-supply in order to determine limitations, in a full-factorial design. As expected, results revealed higher carbon fixation rates for NH 4 + -grown cells compared to growth with NO 3 - under low CO2 conditions. NO 3 - -grown cells accumulated more of the nine analyzed amino acids, especially under Pi-limited conditions, compared to cells provided with NH 4 + . This is probably due to a slower protein synthesis in cells provided with NO 3 - . In contrast to our expectations, compared to NH 4 + -grown cells NO 3 - -grown cells had higher photosynthetic efficiency under Pi-limitation. In conclusion, growth on the Ni-source NH 4 + did not result in a clearly enhanced Ci-assimilation, as it was highly dependent on Pi and CO2 conditions (replete or limited). Results are potentially connected to the fact that C. acidophila is able to use only CO2 as its inorganic carbon (Ci) source.

Inter-site variability of the osteocyte lacunar network in the cortical bone underpins fracture susceptibility of the superolateral femoral neck.

BACKGROUND: The osteocytic lacunar network is considered to be an integral player in the regulation of bone homeostasis, and reduction in osteocytes is associated with reduced bone strength. Here, we analyzed site-specific patterns in osteocyte characteristics and matrix composition in the cortical compartment of the femoral neck to reveal the structural basis of its fragility. METHODS: Cross-sections of the human femoral neck - one of the most common fracture sites - were acquired from 12 female cadavers (age 34-86 years) and analyzed with backscattered scanning electron microscopy and high-resolution micro-computed tomography (µ-CT). The 2D/3D density and size of the osteocyte lacunae as well as bone mineral density distribution (BMDD) were measured in two regions subject to different biomechanical loads in vivo: the inferomedial (medial) region (habitually highly loaded in compression) and the superolateral (lateral) region (lower habitual loading intensity). Using quantitative polarized light microscopy, collagen fiber orientation was quantified in these two regions, accordingly. RESULTS: In 2D measurements, the inferomedial region displayed lower mineralization heterogeneity, 19% higher osteocyte lacunar density (p = 0.005), but equal lacunar size compared to the superolateral region. 3D measurements confirmed a significantly higher osteocyte lacunar density in the inferomedial region (p = 0.015). Osteocyte lacunar density decreased in aged individuals, and inter-site differences were reduced. Site-specific osteocyte characteristics were not accompanied by changes in collagen fiber orientation. CONCLUSIONS: Since osteocyte characteristics may provide valuable insights into bone mechanical competence, the variations in osteocyte properties might reflect the increased fracture susceptibility of the superolateral neck.