Aquarium Experiments Research Articles

Виведення 137Cs з організму карася сріблястого (Carassius gibelio) за різної температурі води у природних умовах Чорнобильської зони відчуження

After the accident at the Chernobyl nuclear power plant, the activity concentration of radionuclides in fish reached hundreds of kBq kg-1. Determination in the dynamics of the content of radionuclides in the body of fish is necessary for the radiation protection of humans and the environment. The data presented in the literature are extremely contradictory and such large differences in the main parameters of radionuclide metabolism in fish require clarification in nature conditions. The aim of this work was to determine the rate of excretion (biological half-life) of 137Cs from the body of silver Prussian carp (Carassius gibelio) at different water temperatures (1-29 °С) depending on the weight of the fish (20±5 and 40±6 g) in real conditions of the Chernobyl Exclusion Zone. As a result of experimental studies, the values of the biological half-life of 137Cs for silver carp was 193-495 days at water temperature 3.7±0.9 °C and 63-92 days at water temperature 22±4 °С were obtained. Due to an increase in the weight of fish (biodilution) at water temperatures > 13 °C, the half-life of the activity concentration of 137Cs in the muscle tissue of fish decreased to 39-58 day-1. The obtained data correspond to the previously obtained values in aquarium experiments. The results obtained in this work show that the level of radioactive contamination of fish in winter will change slightly compared to summer time, which is also confirmed by experimental data obtained in aquarium experiments.

Responses of branching reef corals Acropora digitifera and Montipora digitata to elevated temperature and pCO2.

Anthropogenic emission of CO2 into the atmosphere has been increasing exponentially, causing ocean acidification (OA) and ocean warming (OW). The “business-as-usual” scenario predicts that the atmospheric concentration of CO2 may exceed 1,000 µatm and seawater temperature may increase by up to 3 °C by the end of the 21st century. Increases in OA and OW may negatively affect the growth and survival of reef corals. In the present study, we separately examined the effects of OW and OA on the corals Acropora digitifera and Montipora digitata, which are dominant coral species occurring along the Ryukyu Archipelago, Japan, at three temperatures (28 °C, 30 °C, and 32 °C) and following four pCO2 treatments (400, 600, 800, and 1,000 µatm) in aquarium experiments. In the OW experiment, the calcification rate (p = 0.02), endosymbiont density, and maximum photosynthetic efficiency (Fv/Fm) (both p < 0.0001) decreased significantly at the highest temperature (32 °C) compared to those at the lower temperatures (28 °C and 30 °C) in both species. In the OA experiment, the calcification rate decreased significantly as pCO2 increased (p < 0.0001), whereas endosymbiont density, chlorophyll content, and Fv/Fm were not affected. The calcification rate of A. digitifera showed greater decreases from 30 °C to 32 °C than that of M. digitata. The calcification of the two species responded differently to OW and OA. These results suggest that A. digitifera is more sensitive to OW than M. digitata, whereas M. digitata is more sensitive to OA. Thus, differences in the sensitivity of the two coral species to OW and OA might be attributed to differences in the endosymbiont species and high calcification rates, respectively.

Lacticaseibacillus casei in Diet of Juvenile Convict Cichlid Fish (Amatitlania nigrofasciata): Evaluating Growth Performance, Digestive Enzyme Activities, Immune Responses, and Stress Resistance.

In this study, growth performance, body composition, digestive enzymes activity, mucosal and immunological parameters, cultivable bacterial populations, and stress resistance were investigated in juvenile convict cichlid fish (Amatitlania nigrofasciata) that received a dietary supplement containing 0 (control), 5×106 (LC1), 5×107 (LC2), and 5×108CFUg-1 diet (LC3) Lacticaseibacillus casei PB-LC39. Two hundred and forty fish (2.44±0.04g) were assigned to twelve experimental aquariums and feed ad libitum three times a day for 8weeks. After final sampling, final body weight, percentage of weight gain (WG %), specific growth rate (SGR), food conversion ratio (FCR), and protein content of whole-body composition were significantly higher (P<0.05) in fish fed LC1 diet than other treatments. Total protease, amylase, and lipase activities were also significantly higher (P<0.05) in fish fed LC1 diet than other groups. Total protein from serum and mucus, lysozyme activity, total immunoglobulin (Ig), and serum globulin were significantly increased (P<0.05) in fish fed LC1 diet compared with other groups. Moreover, total counts of lactic acid bacteria (LAB) in fish gut were significantly higher (P<0.05) by different levels of L. casei PB-LC39 than the control group. Recovery rates of fish fed the probiotic cells, after an air-dive test, was significantly increased (P<0.05) compared with the control group. Therefore, the results showed that L. casei PB-LC39 resulted in improving growth, health status, and stress resistance of fish during the rearing of juvenile convict cichlid fish.

Male reproductive strategies in two species of spider crabs, Leurocyclus tuberculosus and Libinia spinosa

Reproductive strategies vary according to season length, individual reproductive traits, and factors associated with the social context such as density and sex ratio of the individuals. The social context predicts the intensity of sexual selection due to its effect on mate choice and intra-sexual competition. Moreover, it exacerbates the costs derived from sexual conflict and allows for the existence of alternative reproductive tactics to counteract such costs. However, these mechanisms are still underexplored in many animal taxa. Majoidea “spider crabs” show a diversity of life history, and behavioral and morphological adaptations for reproduction, which have evolved as an outcome of competition to maximize reproductive success. We aimed to analyze in this study, male reproductive strategies through mating behavior under different social contexts, density of individuals, and males size differences in two species of “spider crabs”, Leurocyclus tuberculosus and Libinia spinosa. The experiments were as follows: one female and one male per aquarium (Experiment 1); a female and three males of similar sizes per aquarium (Experiment 2); and a female and three males of distinct sizes per aquarium (Experiment 3). The experiments showed that mating behavior and duration of mate guarding differ between species and among different social contexts. The presence of males with similar or different sizes showed a relevant different behavior, the female copulated with more than one male when males were similar in size (E2), and copulated only with the largest male when males were different in size (E3). In mate guarding, the shape and the use of the chelipeds had an important role: the male of Leurocyclus tuberculosus grasped the female with the chelipeds and did not fight with other males, but the male of Libinia spinosa surrounded the female with the chelipeds and fought with other males. The longest duration of mate guarding was during precopulatory guarding. Precopulatory guarding was present in all the experiments and its duration did not present significant differences among the experiments. However, in both species, the experiments differ in the duration of postcopulatory guarding until spawning. Gonopod morphology revealed important differences between species and are probably associated with sperm competition. The experiments under distinct social contexts linked to the use and shape of the chelipeds and the morphology of the gonopods, could allow to understand the functional significance of the different reproductive strategies.

Sea urchin repelling Tannin– FeIII complex coating for ocean macroalgal afforestation

Intense seaweed grazing by sea urchins has destroyed kelp forests and accelerated the transformation of these forests into barren areas known as urchin barrens. Once the sea urchins occupy the barren ground, it becomes more challenging to restore the kelp forests. Although phlorotannin, a primary herbivore defense chemical secreted by kelp, has been reported to discourage feeding activities of marine herbivores but the direct application of naturally extracted phlorotannin does not effectively repel sea urchins. In this study, we applied a simple and green Tannin-FeIII (TA-FeIII) coating on substrates as a sea urchin repellent using a cheap, ecofriendly tannin (TA) obtained from biomass as an alternative to phlorotannin. In a model aquarium experiment, most of the sea urchins (Anthocidaris crassispina) in the tank evaded the TA-FeIII-coated substrates. In field tests with 300 sea urchins, the majority of sea urchins could not crawl over the TA-FeIII-coated rope for more than 2 h in contrast to the control group. Hence, the safety, cost-effectiveness, and scalability of the TA-FeIII coating make it a practical candidate to protect the kelp ecosystem from sea urchins.

Increasing confidence for discerning species and population compositions from metabarcoding assays of environmental samples: case studies of fishes in the Laurentian Great Lakes and Wabash River

Community composition data are essential for conservation management, facilitating identification of rare native and invasive species, along with abundant ones. However, traditional capture-based morphological surveys require considerable taxonomic expertise, are time consuming and expensive, can kill rare taxa and damage habitats, and often are prone to false negatives. Alternatively, metabarcoding assays can be used to assess the genetic identity and compositions of entire communities from environmental samples, comprising a more sensitive, less damaging, and relatively time- and cost-efficient approach. However, there is a trade-off between the stringency of bioinformatic filtering needed to remove false positives and the potential for false negatives. The present investigation thus evaluated use of four mitochondrial (mt) DNA metabarcoding assays and a customized bioinformatic Bioinformatic pipeline to increase confidence in species identifications by removing false positives, while achieving high detection probability. Positive controls were used to calculate sequencing error, and results that fell below those cutoff values were removed, unless found with multiple assays. The performance of this approach was tested to discern and identify North American freshwater fishes using lab experiments (mock communities and aquarium experiments) and processing of a bulk ichthyoplankton sample. The method then was applied to field environmental (e) DNA water samples taken concomitant with electrofishing surveys and morphological identifications. This protocol detected 100% of species present in concomitant electrofishing surveys in the Wabash River and an additional 21 that were absent from traditional sampling. Using single 1 L water samples collected from just four locations, the metabarcoding assays discerned 73% of the total fish species that were discerned during four months of an extensive electrofishing river survey in the Maumee River, along with an additional nine species. In both rivers, total fish species diversity was best resolved when all four metabarcoding assays were used together, which identified 35 additional species missed by electrofishing. Ecological distinction and diversity levels among the fish communities also were better resolved with the metabarcoding assays than with morphological sampling and identifications, especially using all four assays together. At the population-level, metabarcoding analyses targeting the invasive round goby Neogobius melanostomus and the silver carp Hypophthalmichthys molitrix identified all population haplotype variants found using Sanger sequencing of morphologically sampled fish, along with additional intra-specific diversity, meriting further investigation. Overall findings demonstrated that the use of multiple metabarcoding assays and custom bioinformatics that filter potential error from true positive detections improves confidence in evaluating biodiversity.

Dataset on body weight and length of rainbow trout, Oncorhynchus mykiss, fed with dihydroquercetin, arabinogalactan or a mixture of both in an aquaria experiment.

The use of natural dietary supplements in aquaculture has received a great deal of attention in recent years. This article provides data describing body weight and length of rainbow trout juveniles fed with natural dietary supplements dihydroquercetin, arabinogalactan or a mixture of both in an aquaria experiment. Before feeding trial, rainbow trout were tagged to identify individuals. Fish grown in tanks were fed one of four diets in duplicate: a basal diet without any supplements (control diet) or a basal diet supplemented with dihydroquercetin (experimental diet 1), arabinogalactan (experimental diet 2) or a mixture of both (experimental diet 3). Our dataset could be used to evaluate the effect of dihydroquercetin, arabinogalactan or a mixture of both on the growth performance of cultivated rainbow trout.

Context-dependency of eelgrass-clam interactions: implications for coastal restoration

Facilitative interactions between co-occurring species sustain diverse communities and constitute a vital functional component of coastal marine ecosystems. In seagrass ecosystems, facilitation ensures the survival and resilience of this important habitat. As seagrass meadows are in decline, innovative restoration strategies incorporating facilitative interactions could open new avenues in marine restoration. Here, we investigated the interactions between eelgrass Zostera marina and the Baltic clam Macoma balthica, and tested whether clams could enhance early survival and biomass increase of transplanted eelgrass shoots in the northern Baltic Sea. We measured eelgrass responses to differing densities of clams, as well as porewater ammonium (NH4+) and phosphate (PO43-) concentrations in field and aquarium experiments. Overall, survival of transplanted plots was high, independent of clam density. Specifically, we found that clams facilitated eelgrass above- and below-ground biomass in low porewater nutrient conditions, potentially through nutrient release, but inhibited growth in high-nutrient conditions, particularly where clams were added at high densities. Our results show the important role of infaunal bivalves for nutrient fluxes within seagrass meadows. Most notably, we highlight the importance of considering and testing context- and density-dependency when studying interspecific interactions, as clams could both benefit and hamper Zostera biomass increase. This becomes particularly crucial when incorporating such interactions in a restoration context.

Shading by marine litter impairs the health of the two Indo-Pacific scleractinian corals Porites rus and Pavona cactus

Marine litter is recognized as one factor affecting coral health. It causes shading, bleaching, physical damage, necrosis, and mortality. This study provides the first evidence that direct contact by plastic and cotton affects coral health within 60 days. In a controlled aquarium experiment, two common Indo-Pacific scleractinian corals, Porites rus and Pavona cactus, were shaded for 60 days by transparent plastic (polypropylene, PP), dark plastic (PP) and cotton. Cotton disintegrated completely after 30 to 42 days, allowing the corals to recover. Transparent plastic became opaque over time due to microfouling, resulting in bleaching of the affected coral parts. Dark plastic had the strongest effect, including bleaching, necrosis and reduced growth within 60 days. Moreover, the two coral species responded differently to the treatments. This is the first report demonstrating that plastic and cotton litter can affect coral health and even cause partial mortality within 60 days.

The Influence of Copper Ions on Cellular Elements of the Coelomic Fluid of Starfish Asterias Rubens L.

This paper is devoted to studying the effect of copper ions on the number and ratio of various subpopulations and functional properties of cells in the coelomic fluid of the starfish Asterias rubens. The experiments were carried out at the White Sea Biological Station named after N.A. Pertsov of the Faculty of Biology of Moscow State University. Starfish were exposed for 96 h to copper chloride (II) in concentrations of Cu2 + ions 0.78, 1.95, and 3.91 μM in aquariums. The number of coelomocytes was significantly increased in starfish kept in aquariums with a maximum concentration of copper ions. The distribution of cell subpopulations was also changed. The proportion of small cells increased from 9 to 15.5%, that of agranulocytes increased from 61 to 75%, and that of granulocytes decreased from 30 to 8.5%. Expression of stress-induced proteins 70 (HSC70/HSP70 ) that was determined by Western blot analysis increased in starfish in all experimental aquariums. The viability of the isolated coelomocytes estimated by the absorption of neutral red (NR) dye increased in starfish in experimental aquariums with copper ion concentrations of 0.78 and 1.95 μM as compared with the control. However, at the concentration of 3.91 μM, NR absorption drastically decreased. Taken together, it was shown that A. rubens responded to the exposure to copper (II) ions by increasing the number of circulating coelomocytes and enhancing the proportion of phagocytes subpopulation. It presumably may serve as a compensatory mechanism in response to the toxic effect of copper (II).

Can environmental DNA be used to detect first arrivals of the cane toad, Rhinella marina, into novel locations?

AbstractEradicating invasive species is difficult, but success is more likely when populations are small after arrival. The cane toad, Rhinella marina, is an invasive pest species that threatens native fauna worldwide. Increasingly, environmental DNA (eDNA) is used as a technique to monitor the presence of invasive species given its power to detect low numbers of individuals. We aimed to investigate eDNA persistence in freshwater at three different temperatures (25, 30 and 35°C) and eDNA detection thresholds for R. marina using controlled experiments in aquaria. For the latter, two water volumes and two cane toad exposure times were used (800 or 200 L volume with 5 or 30 min exposure). A 15‐ml water sample was collected from each replicated aquaria and preserved with 5 ml Longmire's buffer. Environmental DNA was extracted and four technical quantitative PCR replicates were analyzed targeting the cane toad 16S rDNA mitochondrial gene. Environmental DNA decayed rapidly in water and was reliably detected for up to 3 days after cane toad removal, regardless of the temperature treatment. Also, cane toad eDNA was detected in the water after a 5‐min initial exposure of a single individual in 800 L of water. Under the physical parameters tested here, a positive detection means that a cane toad has been in contact with the water body between 1 and 3 days prior to the sampling event. The results of the present study show the importance of eDNA for determining the presence of a species that occurs at low abundance in a small water body, such as at the onset of a cane toad invasion.

Organic eutrophication increases resistance of the pulsating soft coral Xenia umbellata to warming.

Recent research indicates that hard corals in a process that is termed phase shift are often replaced by soft corals in reefs. The simultaneous occurrence of local (i.e. organic eutrophication as highly under-investigated parameter) and global (i.e. ocean warming) factors may facilitate these phase shifts as hard corals are negatively affected by both ocean warming and organic eutrophication. Knowledge about soft coral responses to environmental change remains incomplete, although these organisms are becoming important players in reefs. The present study thus investigated the individual and combined effects of organic eutrophication (as glucose addition) and warming on the ecological data of the pulsating soft coral Xenia umbellata. We assessed health status, growth and pulsation rates of soft corals in a 45 day aquarium experiment, with first manipulation of organic eutrophication (no, low, medium and high glucose addition) over 21 days followed by step-wise increases in water temperature from 26 to 32 °C over 24 days. Findings revealed that glucose addition did not affect health status, growth and pulsation rates of the investigated soft corals. Under simulated ocean warming, soft corals that had experienced organic eutrophication before, maintained significantly higher pulsation rates (averaging 22 beats per minute—bpm) and no mortality compared to the controls that showed a decrease of 56% (averaging 15 bpm) in pulsation rates and mortality of 30% at water temperatures of 32 °C compared to 26 °C. This apparently positive effect of organic eutrophication on the ecological data of soft corals under an ocean warming scenario decreased with increasing water temperature. This study thus indicates that (a) organic eutrophication as additional energy source up to a certain threshold may increase the resistance of soft corals to ocean warming and (b) pulsation rates of soft corals may be used as inexpensive, easily detectable, and non-invasive early warning indicator for ocean warming effects on benthic reef communities. When comparing findings of this study for soft corals with previous results for hard corals, it can be assumed that soft corals under the predicted increases of organic eutrophication and warming gain more and more competitive advantages. This may further facilitate phase shifts from hard to soft corals in warming reefs.

Context-dependent agonistic response to neighbours along territory boundaries in a cichlid fish

Abstract In many territorial animals, the level of aggression exhibited toward conspecifics depends on the magnitude of the threat they pose and/or their familiarity. We conducted aquarium experiments to investigate whether the territorial cichlid fish Julidochromis transcriptus has a context-dependent agonistic response to neighbouring conspecifics. The attacking rate of territory-holding males decreased against neighbour males in an adjacent aquarium over time but increased when stranger males replaced the neighbours in the same aquarium, indicating that males displayed the ‘dear enemy effect’ toward familiar neighbours. Females were not aggressive and the dear enemy effect was not apparent among them. Interestingly, territory-holding males also increased their attacking rate against neighbour males shifted to the opposite aquarium. Territory-holding males will modify their agonistic response to conspecific males according to the relative threat of their opponents in the context of the ‘correct-incorrect boundary paradigm’.

Negative impacts of the sea lice prophylactic emamectin benzoate on the survival of hatchery released salmon smolts in rivers

Emamectin benzoate (EB) is a prophylactic pharmaceutical used to protect Atlantic salmon (Salmo salar) smolts migrating out of rivers and into the ocean against sea lice parasites. Randomized control trials comparing the marine survival of smolts treated with EB to a control group is used to calculate the fraction of marine mortality attributable to sea lice parasitism. However, it is assumed that there is no baseline difference in survival induced by the application of EB treatment. We used a combined laboratory and field study approach to investigate the potential impacts of EB treatment on behaviour and survival of hatchery-reared Atlantic salmon in western Norway. In aquaria experiments, EB-treated salmon smolts did not differ significantly in exploratory behaviour. Fish from treated groups responded similarly to simulated predator attack with spontaneous escape and elevated gill beat rate. Three rivers in the Osterfjord system of western Norway were selected for field experiments, Dale, Vosso, and Modalen. Dale River smolts were treated with intraperitoneal EB injections and had lower probability of detection in a wolf trap downstream of the release site than control smolts. Salmon smolts raised in the Vosso River hatchery were treated with EB delivered in their food and were detected on PIT antennas at the rivermouth of Vosso and Modalen at lower rates than control fish, but only when released at downstream sites. Calculation of risk ratios suggested that the bias in mortality caused by treatment with EB decreased the estimated survival of treated fish from an expected 18%to 46%, reducing the observable negative impact of sea lice on Atlantic salmon smolts in randomized control trials. The results suggest that estimates of the fraction of mortality attributable to sea lice may be underestimated due to lower baseline survival of treated fish caused by treatment and bring urgent attention towards a potential systematic underestimation of the impacts of sea lice on wild salmon.

Environmental DNA analysis shows high potential as a tool for estimating intraspecific genetic diversity in a wild fish population.

Environmental DNA (eDNA) analysis has recently been used as a new tool for estimating intraspecific diversity. However, whether known haplotypes contained in a sample can be detected correctly using eDNA-based methods has been examined only by an aquarium experiment. Here, we tested whether the haplotypes of Ayu fish (Plecoglossus altivelis altivelis) detected in a capture survey could also be detected from an eDNA sample derived from the field that contained various haplotypes with low concentrations and foreign substances. A water sample and Ayu specimens collected from a river on the same day were analysed by eDNA analysis and Sanger sequencing, respectively. The 10L water sample was divided into 20 filters for each of which 15 PCR replications were performed. After high-throughput sequencing, denoising was performed using two of the most widely used denoising packages, unoise3 and dada2. Of the 42 haplotypes obtained from the Sanger sequencing of 96 specimens, 38 (unoise3) and 41 (dada2) haplotypes were detected by eDNA analysis. When dada2 was used, except for one haplotype, haplotypes owned by at least two specimens were detected from all the filter replications. Accordingly, although it is important to note that eDNA-based method has some limitations and some risk of false positive and false negative, this study showed that the eDNA analysis for evaluating intraspecific genetic diversity provides comparable results for large-scale capture-based conventional methods. Our results suggest that eDNA-based methods could become a more efficient survey method for investigating intraspecific genetic diversity in the field.

An Eulerian multimaterial framework for simulating high-explosive aquarium tests

Aquarium tests of cylindrical high-explosive charges provide optical data of the detonation front velocity and shape, propagation of the shock wave in the surrounding water, and expansion rates of the detonation products behind the front. Data from aquarium experiments is often used for calibration of reactive burn models based on phenomenological equations of state (EOS) and reaction rate laws. This paper presents a multimaterial numerical modeling framework to solve the 2D axisymmetric reactive Euler equations for high-explosive aquarium tests, in particular for ammonium nitrate - fuel oil (ANFO) explosives. An extension of the Ghost Fluid Method (GFM) is used to handle the dynamic material interfaces for the ANFO explosion products, the charge-confining material (polymethyl methacrylate PMMA), and the surrounding water. This study analyzes the sensitivity of calculations (both computational efficiency and numerical accuracy) to different algorithms for the material interface models including the original GFM versus Riemann solver-based strategies. A novel method for defining the left and right states in the interfacial Riemann problem eliminates the need for sorting or nodal interpolation during the projection along the material interface. Numerical tests indicate that populating the interface node values using the Riemann solution mitigate the overheating error observed in steady-state calculations. Solution convergence and computational efficiency are explored as a function of the spatial and temporal order of the schemes. Results from the computational model with analytical equations of state and fitted reaction rate parameters show very good quantitative agreement with experimentally observed detonation front velocity, reaction products expansion, and shock wave propagation in the surrounding water for a cylindrical ANFO charge encased in PMMA. The proposed modeling framework, in conjunction with experimental tests, provides a reliable tool to assess equations of state and reaction rate expressions for reactive burn models of confined high explosives.

An Untargeted Metabolomic Approach for Microphytobenthic Biofilms in Intertidal Mudflats

Microphytobenthic (MPB) biofilms in intertidal muddy sediments play important ecological functions in coastal ecosystems. These biofilms are mainly composed of epipelic diatoms but also prokaryotes, with a dominance of bacteria, which excrete diverse extracellular polymeric substances (EPS) according to their environment. While numerous studies have investigated the main components of these EPS matrices via traditional colorimetric assays, their fine composition, notably in specialized metabolites, is still largely unknown. A better chemical characterization of these MPB biofilms is necessary, especially regarding the numerous functions their chemical components play for microorganisms (e.g. motility, cell protection, defense mechanisms, chemical communication), but also for coastal systems (e.g. primary production, sediment stabilization, larval settlement of some invertebrates with high economical value). An alternative approach to traditional analyses is the use of untargeted metabolomic techniques, which have not yet been applied to such MPB biofilms. The objectives of the present study were to (a) propose a protocol for metabolic fingerprinting by LC-MS and GC-MS for metabolites analysis in polar and non-polar fractions in MPB biofilms extracted from mudflat sediment and to (b) apply this protocol to a case study: the effect of light exposure on the metabolomic fingerprint of the MPB biofilm community. We compared three extraction methods using different mixes of solvents and selected a methanol/chloroform mix (1:1), which gave better results for both techniques and fractions. We then applied the selected protocol to our case study using a short-term light exposure experiment in aquaria (7 days). The present study is the first using a detailed untargeted metabolomic approach on MPB biofilms from mudflat sediment and will provide a solid baseline for further work in this area

COPPER EFFECT ON COMPONENTS OF THE ANTIOXIDANT SYSTEM OF THE DIGESTIVE GLAND IN THE FRESHWATER MUSSEL ANODONTA CYGNEA

The effects of copper accumulation and its impact on the functioning of some components of the antioxidant system (AOS) – CAT, GST, GSH – in the digestive gland of the freshwater mussel Anodonta cygnea were studied in an aquarium experiment. After a 12‑day acclimation, the mussels were divided among aquariums with Cu²⁺ concentrations of 0, 5, 50, 100, and 250 μg/L. Hepatopancreas tissue samples were taken for analysis after 1, 3, and 7 days. Copper ions content, GST and CAT activity, as well as GSH concentration were determined in the tissue. The accumulation process was shown to be doseand time-dependent. The metal accumulated in this concentration was not lethal. The rate of the metal accumulation in the studied organ increased along with the concentration of copper ions in the water. The activity of AOS enzymes in the swan mussel is modified as an adaptation in response to elevated copper concentrations to neutralize the products of oxidative reactions. Changes in the activity of AOS enzymes (CAT and GST) were observed mainly on the seventh day of the experiment at the highest copper concentrations tested. GST proved to be the most stable enzyme. The GSH level in the digestive gland changed 1 day after the start of the experiment, corroborating this tripeptide is involved in the first wave of protection against xenobiotics. Copper ions have a cumulative effect. The changes in enzyme activity observed on day 7 at Cu²⁺ concentration of 250 μg/L suggest that a longer exposure to copper, even in low concentrations, can be detrimental.

Damselfishes alleviate the impacts of sediments on host corals.

Mutualisms play a critical role in ecological communities; however, the importance and prevalence of mutualistic associations can be modified by external stressors. On coral reefs, elevated sediment deposition can be a major stressor reducing the health of corals and reef resilience. Here, we investigated the influence of severe sedimentation on the mutualistic relationship between small damselfishes (Pomacentrus moluccensis and Dascyllus aruanus) and their coral host (Pocillopora damicornis). In an aquarium experiment, corals were exposed to sedimentation rates of approximately 100 mg cm−2 d−1, with and without fishes present, to test whether: (i) fishes influence the accumulation of sediments on coral hosts, and (ii) fishes moderate partial colony mortality and/or coral tissue condition. Colonies with fishes accumulated much less sediment compared with colonies without fishes, and this effect was strongest for colonies with D. aruanus (fivefold less sediment than controls) as opposed to P. moluccensis (twofold less sediment than controls). Colonies with symbiont fishes also had up to 10-fold less sediment-induced partial mortality, as well as higher chlorophyll and protein concentrations. These results demonstrate that fish mutualisms vary in the strength of their benefits, and indicate that some mutualistic or facilitative interactions might become more important for species health and resilience at high-stress levels.

Reliable eDNA detection and quantification of the European weather loach (Misgurnus fossilis).

The European weather loach (Misgurnus fossilis) is a cryptic and poorly known fish species of high conservation concern. The species is experiencing dramatic population collapses across its native range to the point of regional extinction. Although environmental DNA (eDNA)-based approaches offer clear advantages over conventional field methods for monitoring rare and endangered species, accurate detection and quantification remain difficult and quality assessment is often poorly incorporated. In this study, we developed and validated a novel digital droplet PCR (ddPCR) eDNA-based method for reliable detection and quantification, which allows accurate monitoring of M. fossilis across a number of habitat types. A dilution experiment under laboratory conditions allowed the definition of the limit of detection (LOD) and the limit of quantification (LOQ), which were set at concentrations of 0.07 and 0.14 copies μl-1 , respectively. A series of aquarium experiments revealed a significant and positive relationship between the number of individuals and the eDNA concentration measured. During a 3 year survey (2017-2019), we assessed 96 locations for the presence of M. fossilis in Flanders (Belgium). eDNA analyses on these samples highlighted 45% positive detections of the species. On the basis of the eDNA concentration per litre of water, only 12 sites appeared to harbour relatively dense populations. The other 31 sites gave a relatively weak positive signal that was typically situated below the LOQ. Combining sample-specific estimates of effective DNA quantity (Qe ) and conventional field sampling, we concluded that each of these weak positive sites still likely harboured the species and therefore they do not represent false positives. Further, only seven of the classified negative samples warrant additional sampling as our analyses identified a substantial risk of false-negative detections (i.e., type II errors) at these locations. Finally, we illustrated that ddPCR outcompetes conventional qPCR analyses, especially when target DNA concentrations are critically low, which could be attributed to a reduced sensitivity of ddPCR to inhibition effects, higher sample concentrations being accommodated and higher sensitivity obtained.