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Selective contactless manipulation of organisms with intrinsic mobility from heterogeneous mixture is essential for biomedical engineering and microbiology. Acoustic manipulation, compared to its optical, magnetic, and electrostatic counterparts, provides superior bio-compatibility and additive-free properties. In this study, we present an acoustic manipulation system capable of selectively trapping, translating, rotating, and orienting individual organisms from in-Petri dish organism mixture using a phased transducer array and microscope, by dynamically steering the acoustic field. Specifically, using brine shrimp and zebrafish populations as example, the to-be-manipulated organisms with different sizes or morphologies can be manually designated by the user in microscopic image and interactively localized. Thereafter, the selected organisms can be automatically trapped from the heterogeneous mixture using a multiple focal point-based acoustic field steering method. Finally, the trapped organisms can be translated, rotated, and oriented in regard to the user's distinct manipulation objectives in instant response. In different tasks, closed-loop positioning and real-time motion planning control are performed, highlighting the innovation in terms of automation and accuracy of our manipulation technique. The results demonstrate that our acoustic manipulation system and acoustic field steering method enable selective, stable, precision, real-time, and in-Petri dish manipulation of organisms from heterogeneous mixture.

Diapause exhibited by embryos of Artemia franciscana is accompanied by severe arrest of respiration. A large fraction of this depression is attributable to downregulation of trehalose catabolism that ultimately restricts fuel to mitochondria. This study now extends knowledge on the mechanism by revealing metabolic depression is heightened by inhibitions within mitochondria. Compared with that in embryo lysates during post-diapause, oxidative phosphorylation (OXPHOS) capacity P is depressed during diapause when either NADH-linked substrates (pyruvate and malate) for electron transfer (electron transfer capacity, E) through respiratory Complex I or the Complex II substrate succinate are used. When pyruvate, malate and succinate were combined, respiratory inhibition by the phosphorylation system in diapause lysates was discovered as judged by P/E flux control ratios (two-way ANOVA; F1,24=38.78; P<0.0001). Inhibition was eliminated as the diapause extract was diluted (significant interaction term; F2,24=9.866; P=0.0007), consistent with the presence of a diffusible inhibitor. One candidate is long-chain acyl-CoA esters known to inhibit the adenine nucleotide translocator. Addition of oleoyl-CoA to post-diapause lysates markedly decreased the P/E ratio to 0.40±0.07 (mean±s.d.; P=0.002) compared with 0.79±0.11 without oleoyl-CoA. Oleoyl-CoA inhibits the phosphorylation system and may be responsible for the depressed P/E in lysates from diapause embryos. With isolated mitochondria, depression of P/E by oleoyl-CoA was fully reversed by addition of l-carnitine (control versus recovery with l-carnitine, P=0.338), which facilitates oleoyl-CoA transport into the matrix and elimination by β-oxidation. In conclusion, severe metabolic arrest during diapause promoted by restricting glycolytic carbon to mitochondria is reinforced by depression of OXPHOS capacity and the phosphorylation system., Competing Interests: Competing interests E.G. is founder and CEO of Oroboros Instruments (Innsbruck, Austria)., (© 2024. Published by The Company of Biologists Ltd.)

Ocean acidification and microplastic pollution are two of the major ecological concerns. The distribution of large quantities of plastic debris and microplastics all across the oceans emphasises the need to determine the influence of microplastics in ocean acidification and to evaluate its concomitant toxicological effects on aquatic life forms. Studies on the combined impact of both the stressors are very limited, but much needed in the current scenario. Where most of the present-day research use purchased microplastics of defined size and morphology (microspheres, fibres, rods, etc.), the present study employs prepared "true to life microplastics" that resemble the environmental microplastic pollutants in morphology and size heterogeneity. The present study focusses on evaluating the fate and impact of oceanic microplastics on the physiology and development of Artemia salina (Brine shrimp), one among the most ecologically significant zooplankton species. Natural sea water was acidified by controlled perturbation of carbon dioxide using a valve system. The hatching rate of A. salina cysts receded significantly (p < 0.05) upon singular exposures to microplastics and low pH (7.80), whereas combined effect was insignificant. The reactive oxygen species (ROS) elevated as a result of individual exposures to microplastics and low pH. However, only in 0.5 mg mL -1 PE treatments at pH 7.80, an additive impact was reported for ROS activity (p < 0.05). The SOD activities increased significantly but it can be attributed as the individual responses towards exposure to both the stressors. A significant additive impact was not observed for SOD activity (p > 0.05). But during the development, significant morphological anomalies were observed. Changes in the appendages of nauplii and juveniles as a result of combined exposure to microplastics and low pH treatments are significant findings. Our observations suggest that coupled exposure to microplastics and low pH could induce significant oxidative stress in the marine zooplanktons and also adversely affect their normal development. Findings from the current study emphasise the need for further research to understand the coupled toxicological impacts of ocean acidification and predominant pollutants such as microplastics to other marine animals as well., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

A great diversity of crustacean zooplankton found in inland and coastal waters produce embryos that settle into bottom sediments to form an egg bank. Embryos from these banks can remain dormant for centuries, creating a reservoir of genetic diversity. A large body of literature describes the ecological and evolutionary importance of zooplankton egg banks. However, literature on the physiological traits behind dormancy in crustacean zooplankton are limited. Most data on the physiology of dormancy comes from research on one species of anostracan, the brine shrimp, Artemia franciscana. Anoxia-induced dormancy in this species is facilitated by a profound and reversible acidification of the intracellular space. This acidification is accompanied by a reversible depletion of adenosine triphosphate (ATP). The present study demonstrates that acidification of the intracellular space also occurs in concert with a depletion of nucleoside triphosphates (NTPs) in the Antarctic copepod, Boeckella poppei. Like A. franciscana, the depletion of NTPs and acidification are rapidly reversed during aerobic recovery in B. poppei. These data provide the first comparative evidence that extreme dormancy under anoxia in crustacean zooplankton is associated with intracellular acidification and an ability to recover from the depletion of ATP., (© 2023. Springer Nature Limited.)

The use of small aquatic model organisms to investigate the behavioral effects of chemical exposure is becoming an integral component of aquatic ecotoxicology research and neuroactive drug discovery. Despite the increasing use of invertebrates for behavioral phenotyping in toxicological studies and chemical risk assessments, little is known regarding the potential for environmental factors-such as geometry, size, opacity and depth of test chambers-to modulate common behavioral responses. In this work, we demonstrate that test chamber geometry, size, opacity and depth can affect spontaneous, unstimulated behavioral responses of euryhaline crustacean Artemia franciscana first instar larval stages. We found that in the absence of any obvious directional cues, A. franciscana exhibited a strong innate wall preference behavior. Using different test chamber sizes and geometries, we found both increased wall preference and lowered overall distance traveled by the test shrimp in a smaller chamber with sharper-angled vertices. It was also determined through quantifiable changes in the chambers' color that the A. franciscana early larval stages can perceive, differentiate and react to differences in color or perhaps rather to light transmittance of the test chambers. The interaction between innate edge preference and positive phototaxis could be consistently altered with a novel photic stimulus system. We also observed a strong initial preference for depth in A. franciscana first instar larval stages, which diminished through the acclimatization. We postulate that the impact of test chamber designs on neurobehavioral baseline responses warrants further investigation, in particular considering the increased interest in behavioral eco-neurotoxicology applications., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Alkyl-PAHs are the predominant form of PAHs in crude oils which are supposed to demonstrate different toxicities compared to non-alkyl PAHs. Little information is available about the toxicity of alkyl-PAHs on marine Artemia. This study addressed and compared the lethal, behavioral, growth and developmental toxicities of three alkyl-PAHs, namely 3-methyl phenanthrene (3-mPhe), retene (Ret) and 2-methyl anthracene (2-mAnt), to their non-alkyl forms, phenanthrene (Phe) and anthracene (Ant) using Artemia parthenogenetica (nauplii, <24 h) as test organism following a 48 h and a 7 d of exposure, respectively. Benzo-a-pyrene (Bap) was selected as a reference toxicant for the comparison with the above alkyl-PAHs and non-alkyl PAHs. Results showed that for all tested endpoints, A. parthenogenetica nauplii had the highest sensitivity to Bap while Ant had no significant effect on nauplii survival or development within given concentrations. Considering the aqueous freely dissolved PAH concentrations, the 48 h-LC 50 (survival), 48 h-EC 50 (immobility) and 7 d-LC 10 (survival) of Bap were calculated as 0.321, 0.285 and 0.027 μg/L, respectively, which were twofold to fivefold lower than those of Phe, 3-mPhe, Ret, Ant and 2-mAnt. A higher acute toxicity of alkyl-PAHs (3-mPhe and 2-mAnt) than their non-alkyl forms (Phe and Ant) was observed. Not limited to Phe, the common non-polar narcotic mode of action was also observed for Bap, 3-mPhe, Ret and 2-mAnt, which was evident by the inhibited mobility of nauplii. The decreased body lengths were found for all PAH treatments compared to the solvent control, whereas instar retardations were only found in nauplii exposed to Bap, Phe and Ret. Our findings emphasized the sensitivity differences of A. parthenogenetica nauplii to selected alkyl PAHs and non-alkyl PAHs and confirmed the application of lethal, behavioral and growth indicators in the toxicity evaluation of selected PAHs other than Ant. However, the distinct toxicities of these PAHs suggested other toxic modes of action may play more important roles apart from narcotic mode of action and need to be elucidated in future studies. In addition, a strong correlation between the body length and the instar of A. parthenogenetica nauplii was observed for each PAH exposure, suggesting that body length can be representative for both growth and developmental indicators during biological monitoring of PAH pollution in marine environment., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Glyphosate-based formulations are the most commonly used herbicides worldwide with the risk of potential contamination of aquatic bodies. The present study assessed the response of four marine crustaceans to three different brands of herbicides Roundup®Platinum, Efesto® and Taifun® MK CL.T, under two selected temperatures of 20 °C and 30 °C. The harpacticoid copepod Tigriopus fulvus, the anostracan Artemia franciscana, the amphipod Corophium insidiosum and the isopod Sphaeroma serratum were chosen as testing organisms. Effects of herbicides and temperatures were assessed by estimating lethal concentrations. The results showed that the high temperature rises the toxicity of glyphosate with an increase of mortality of all the tested species. This is an important aspect for future risk assessments of pesticides under global climate change scenarios. Efesto® resulted the most toxic brand, showing C. insidiosum the most sensitive with 96 h-LC50 values of 3.25 mg/L acid equivalent (a.e.) at 30 °C and 7.94 mg/L a.e. at 20 °C followed by T. fulvus while A. franciscana and S. serratum were the less sensitive. This study provides important information for assessing the toxic effects of three different brands of glyphosate-based herbicides on non-target marine organisms suggesting that they should be carefully managed to minimize any negative impact on marine organisms., (Published by Elsevier Inc.)

The hatchability, mortality rate, lipid peroxide levels, and swimming speed of Artemia salina have been compared based on short exposures of ZnCl 2 , CdCl 2 , and HgCl 2 in artificial seawater. The hatching tests were carried out for 12, 24, 36, and 48 h at 28 °C. Mortality rate and lipid peroxide (LPO) levels were determined after 24 h of exposure at 28 °C, in the dark, and on living larvae using the FOX method. The swimming speed was determined after 24 h using a microcomputer coupled to a digital camera, with simultaneous treatment of the recorded images every 25 s, at 25 °C, under red-light irradiation. Results showed that Zn caused a gradual inhibition of the hatching for concentrations <900 µmol L -1 ; however, Cd and Hg displayed almost complete inhibition for concentrations ≤100 µmol L -1 . Also, the heavy metals caused a dose-dependent increase of mortality (LD50) in the following order: Zn = 3290 µmol L -1 < Cd = 2206 µmol L -1 < Hg = 15.6 µmol L -1. Furthermore, significant LPO levels were found for Cd (1500-2000 µmol L -1 , p < 0.001) and Hg (5-20 µmol L -1 , p < 0.001). Finally, the swimming speed values increased significantly, for Zn ≈ 2.5 mm s -1 (1500 µmol L -1 , p < 0.001), Cd ≈ 3.5 mm s -1 (2000 µmol L -1 , p < 0.05), and Hg ≈ 4.0 mm s -1 (15 µmol L -1 , p < 0.05), after 24 h exposure. There is a clear dose-dependent toxicity, indicating that Zn, Cd and Hg can induce significant changes in hatchability, mortality, and ethological and biochemical parameters., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Artemia salina cysts collected from the Sebkha of Sidi El Hani hatched and cultivated in the laboratory were investigated. Nauplii were acclimated to laboratory conditions until reproductively mature (Generation 1; G1). Reproductive performance in terms of total cysts and nauplii offspring, days between brood production, and cyst and nauplii numbers was evaluated. Nauplii produced by specimens in the G1 were defined as those of G2 with there also being third (G3) and fourth (G4) generations. The aims were to evaluate Artemia salina in aquaculture conditions with the long-term aim of developing a system for "artemiculture". There was a larger number of cysts or nauplii per brood between G1 (60.7 ± 10.9 nauplii or cysts/female) and subsequent generations, notably G2 (122 nauplii or cysts/female, P <  0.05). The mean number of cysts and nauplii per female increased from the first and last brood. Number of cysts produced per female increased markedly (P <  0.05) from G1 (54.5 ± 14.2) to G3 (128.9 ± 39.2). Percentage of females producing nauplii increased from G1 (20 %) to G4 (100 %). There was, therefore, an increase in percentage of ovoviviparous offspring (nauplii) per female (P <  0.05) from G1 (7%) to G4 (71 %). There were no differences, however, between mean numbers of broods per female, and numbers of days between broods. The results indicate there are variations in reproduction from oviparity to ovoviviparity as consecutive generations of Artemia salina are cultivated. In summary, there was a rapid adaptation of Artemia salina of Sidi El Hani to laboratory conditions., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Reef-building corals rely on both heterotrophy and endosymbiotic dinoflagellate autotrophy to meet their metabolic needs. Those looking to culture these organisms for scientific or industrial purposes must therefore consider both feeding regimes and the light environment. Herein the effects of three photosynthetically active radiation (PAR) levels were assessed in fed and unfed specimens of the model coral Pocillopora acuta that were cultured in a recirculating aquaculture system (RAS). Half of the corals were fed Artemia sp. brine shrimp in a separate feeding tank to prevent biofouling, and fragments were exposed to PAR levels of 105, 157, or 250 μmol quanta m -2  s -1 over a 12-h period each day. All cultured corals survived the 140-day treatment, and the physiological response variables assessed-buoyant weight, specific growth rate, linear extension, color, and Fv/Fm-were significantly influenced by feeding, and, to a lesser extent, light. Specifically, fed corals grew faster and larger, and presented darker pigmentation; corals fed at the highest light levels grew at the fastest rate (6 cm year -1 or 175 mg g -1  week -1 ). Given the high physiological performance observed, we advocate the active feeding of brine shrimp in RAS by those looking to cultivate P. acuta, and likely other corals, over long-term timescales.

In their role as molecular chaperones, heat shock proteins (Hsps) mediate protein folding thereby mitigating cellular damage caused by physiological and environmental stress. Nauplii of the crustacean Artemia franciscana respond to heat shock by producing Hsps; however, the effects of cold shock on Hsp levels in A. franciscana have not been investigated previously. The effect of cold shock at 1 °C followed by recovery at 27 °C on the amounts of ArHsp90, Hsp70, ArHsp40, and ArHsp40-2 mRNA and their respective proteins in A. franciscana nauplii was examined by quantitative PCR (qPCR) and immunoprobing of western blots. The same Hsp mRNAs and proteins were also quantified during incubation of nauplii at their optimal growth temperature of 27 °C. qPCR analyses indicated that the abundance of ArHsp90, Hsp70, and ArHsp40 mRNA remained relatively constant during both cold shock and recovery and was not significantly different compared with levels at optimal temperature. Western blotting revealed that ArHsp90, ArHsp40, and ArHsp40-2 were generally below baseline, but at detectable levels during the 6 h of cold shock, and persisted in early recovery stages before declining. Hsp70 was the only protein that remained constant in quantity throughout cold shock and recovery. By contrast, all Hsps declined rapidly during 6 h when nauplii were incubated continuously at 27 °C optimal temperature. Generally, the amounts of ArHsp90, ArHsp40, and ArHsp40-2 were higher during cold shock/recovery than those during continuous incubation at 27 °C. Our data support the conclusion that low temperature preserves Hsp levels, making them available to assist in protein repair and recovery after cold shock.

Quantum dots (QDs), such as cadmium selenide (CdSe) and lead selenide (PbSe) exhibit excellent optical, magnetic and chemical properties due to their extremely size (ca. 1-10 nm) and are attractive semiconductor nanomaterials for optical studies and energy storage. In this study, aqueous synthesis of CdSe and PbSe QDs in a size range of 2-10 nm was described. Synthesized QDs were characterized using SEM and TEM, DLS, zeta potential, FTIR, EDX and XRD. Highest accumulation (72.5 ± 5.8 mg L -1 ) of PbSe QDs occurred at 10 ppm suspensions. In general accumulation increased up to 48 h exposure then fluctuate tended to decline. For CdSe QDs, accumulation tended to decrease for 72 h exposure except that for 5 ppm groups. For the elimination period, in general, the elimination levels of PbSe and CdSe QDs from exposed individuals decreased (p < 0.05) even it has some fluctuate.

Magnetic silver nanoparticles (MNPAg) are interesting nanotechnology materials with borderless environmental science, that can be used to disinfect water contaminated with pathogenic bacteria. The use of MNPAg leads to increased risk of nanomaterial contamination in the environment, especially natural water sources, with harmful effects on the ecosystem. This study investigating survival and enzyme activity of magnetic O-carboxymethylchitosan loaded silver nanoparticle on Artemia salina. The results showed that mortality increased with increasing concentrations of MNPAg. O-Carboxymethylchitosan loaded silver nanoparticles were found to be more toxic, with a LC 50 of 902.1 mg/L for γ-Fe 2 O 3 /Ag without reducing agent. Accumulation of silver on Artemia salina depends on the type of nanoparticle. Accumulation of nanoparticle containing polymers (carboxymethylchitosan/γ-Fe 2 O 3 /Ag without reducing agent, carboxymethylchitosan/γ-Fe 2 O 3 /Ag reduced with sucrose and carboxymethylchitosan/γ-Fe 2 O 3 /Ag reduced with NaBH 4 ) were found to be higher than γ-Fe 2 O 3 /Ag reduced with NaBH 4 , γ-Fe 2 O 3 /Ag reduced with sucrose and γ-Fe 2 O 3 /Ag without reducing agent under the same experimental conditions. The antioxidant enzyme (CAT, SOD and GST) activities increased slightly following exposure, indicating that the toxic effects are related to oxidative stress. The combined results so far indicate that MNPA does not have the potential to affect aquatic organisms when released into the ecosystem., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Nanoplastics (NPs) have become one of the most serious environmental problems nowadays. The environmental issues linked to NPs are attributed to the effects after ingestion in marine organisms. Due to the incipient and controversial information about the effects of PS NPs on the feeding of organisms, the aim of this work is to assess (i) digestion dynamics of Artemia franciscana when exposed to PS NPs as the lowest concentration of PS NPs reported in toxicity test [0 (control), 0.006 and 0.6 mg·L-1] and possible interferences in the ingestion of microalgae and (ii) the accumulation and depuration of PS NPs by A. franciscana. Artemia were subjected to ingestion experiments [24 h and 3.5 h], in which the organisms were exposed to PS NPs or to PS NPs + microalgae. Post-exposure feeding (24 h exposure and 2 h feeding) and depuration (24 h exposure and 24 h of depuration) were also carried out. More than 90% of the PS NPs were ingested by Artemia and bioaccumulated in the mandible, stomach, gut, tail gut and appendages after 24 h. The ingestion of microalgae was not affected by the presence of the PS NPs. Data of post-exposure feeding indicated that Artemia previously exposed to plastic and/or microalgae presented similar microalgal ingestion (around 70%); the highest microalgal consumption (around 90%) was recorded in the treatment in which Artemia were previously starved (no plastic and no microalgae). The presence of PS NPs in the gut after the depuration experiments indicates that 24 h was not enough to eliminate the PS NPs., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Toxicities of the marine algae Alexandrium minutum and its excreted gonyautoxins (GTXs) to the marine crustacean Artemia salina were investigated. Mortality was observed for neither larvae nor adult A. salina exposed to A. minutum at a density of 5000 cells/mL or 0.5 μM GTX2/3. After exposure, the full transcriptome of adult A. salina was assembled and functionally annotated. A total of 599,286 transcripts were obtained, which were clustered into 515,196 unigenes. Results of the transcriptional effect level index revealed that direct exposure to the toxic algae A. minutum caused greater alterations in the transcriptome than did exposure to the extracellular product GTX2/3. Mechanisms of effects were different between exposure of A. salina to A. minutum cells or GTX2/3. Exposure to A. minutum modulated formation of the ribonucleoprotein complex and metabolism of amino acids and lipids in A. salina. Exposure to GTX2/3 exposure inhibited expression of genes related to metabolism of chitin, which might result in disruption of molting process or disturbed sheath morphogenesis. Overall, effects on transcription observed in this study represent the first report based on application of next generation sequencing techniques to investigate the transcriptomic response of A. salina exposed to an environmentally realistic level of A. minutum or GTX2/3., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The use of vegetable waste and its screening for potential cytotoxicity is of utmost importance to ensure its safe use in the feed industry for fish and other animals. We evaluated aqueous and ethanolic extracts of cooked Araucaria angustifolia seed coats. The Stiasny index for the aqueous and ethanolic extracts was 2.87% ± 0.03% and 60.53% ± 4.79%, respectively. Condensed tannins were 11-fold higher in the ethanolic extract than the aqueous extract. The flavonoid and polyphenol contents were 1.7- and 1.8-fold higher in the ethanolic extract than in the aqueous extract, respectively. The 36 h EC 50 for brine shrimp hatchability was 300.32 µg/mL for the aqueous extract, and 76.60 µg/mL for the ethanolic extract. The 24 h LC 50 was 1405.96 µg/mL for the aqueous extract, and it was 356.32 µg/mL for the ethanolic extract. The aqueous extract was nontoxic to A. salina nauplii, and therefore, it can be used as a possible food additive in fish feed. The results also demonstrated that the different solvents used in the extraction affected the yield and the total phenolic, total flavonoid, and condensed tannin content. Further in vivo and cell line cytotoxicity testing is recommended to substantiate these findings.

Environmental monitoring has demonstrated widespread occurrence of the flame-retardant tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), raising concerns about the impact on aquatic life. Using 1 H NMR and GC-MS metabolomics and 20-day body length experiments, we have determined that exposure to TDCIPP affects Artemia franciscana. The LC 50 for a 48 h TDCIPP exposure was determined to be 37.1 ± 1.3 μM. Acute exposure (48 h) to 20.0 μM did not affect A. franciscana body length but did elicit a metabolic change. Chronic exposure to 0.50 μM TDCIPP caused decreased body length in A. franciscana exposed for 20 days and elicited a metabolic response. Principal component analysis revealed variance between acute and chronic exposure along PC1 (36.4%) and between control and TDCIPP along PC2 (17.4%). One-way ANOVA indicated that 19 metabolites were significantly affected by TDCIPP exposure; namely metabolites of the osmolyte class, including betaine, phosphocholine, gadusol, taurine, glycerol and trehalose - metabolites that are essential osmoprotectants in extremophile species. Other pathways that may be perturbed by TDCIPP exposure include one carbon, glycine, serine, threonine, and glycerophospholipid metabolism., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Micro- (<5 mm) and nanoplastics (<1 μm) are emerging threats for marine ecosystems worldwide. Brine shrimp Artemia is recognized as a suitable model among planktonic species for studying the impact of polystyrene nanoparticles (PS NPs) through short and long-term bioassays. Our study aims to evaluate the time-dependent effects of cationic amino-modified PS-NH 2 (50 nm) in A. franciscana after short- (48 h) and long-term exposure (14 days). For this purpose, nauplii were exposed to a concentration range of PS-NH 2 (0.1, 1, 3 and 10 μg/mL) in natural sea water (NSW), and physiological, biochemical and molecular responses were investigated. Short-term exposure to PS-NH 2 caused a decrease in nauplii growth and affected the development in a concentration-dependent manner, long-term exposure impaired the survival, but not the growth and feeding behavior. Oxidative stress was detected after short term exposure as the decrease in the activity of antioxidant enzymes, and was fully evident in the long-term as lipid peroxidation, suggesting an accumulative effect. The decrease in Cholinesterase (ChE) activity observed indicates possible neurotoxic action of PS-NH 2 . Also, Carboxylesterase (CbE) inhibition by PS-NH 2, described for the first time in this study, anticipates potential effects in biotransformation of exogenous and endogenous compounds, being the crustacean juvenile hormone methyl farnesoate (MF) that regulates development and molting, one candidate. Furthermore, short- and long-term exposure to PS-NH 2 affect the expression of genes involved in cell protection, development and molting. Overall, our results reveal that low PS-NH 2 concentrations induce physiological, biochemical and molecular (changes in gene expression) alterations in Artemia, and point at their potential risk for this model organism, supporting the general concern about nanoplastics occurrences in aquatic environments and their ability to represent an ecological threat for aquatic zooplanktonic species., (Copyright © 2019 Elsevier B.V. All rights reserved.)

As with many biological control agents, generalist predators rarely survive prolonged periods of prey scarcity. Towards improving crop establishment of two major predators used in North America, Orius insidiosus and Dicyphus hesperus, this study examined the role of supplemental foods in achieving greater predator survival and faster development. In controlled environment trials, developmental time and survival were compared for predators offered diets including Ephestia eggs, Artemia cysts, Typha pollen, or combinations of these. Nymphal developmental time was significantly shorter and survival greater for both predators reared on diets that included Ephestia eggs. Interestingly, D. hesperus could successfully complete nymphal development on Artemia cysts whereas O. insidiosus could not, alluding to fundamental physiological differences between these predators. In greenhouse assays, D. hesperus was more abundant after six weeks when offered diets that included Ephestia eggs either alone or in combination with pollen or Artemia cysts relative to other diets. In contrast, only diets of Ephestia eggs, Typha pollen or their combination could significantly increase O. insidiosus crop abundance relative to the unfed control. Together, this work highlights important differences in the relative values of supplemental foods for generalist predators used in crop protection. It is also meaningful in guiding biocontrol practitioners globally in the rapidly growing sector of greenhouse vegetable production.

Embryos of the crustacean Artemia franciscana develop either ovoviviparously or oviparously, yielding swimming larvae (nauplii) or encysted gastrulae (cysts), respectively. Nauplii moult several times and become adults whereas cysts enter diapause, a state of dormancy characterized by exceptionally low metabolism and high stress tolerance. Synthesis of molecular chaperones such as the J-domain proteins ArHsp40 and ArHsp40-2 occurs during embryo development and post-diapause growth of A. franciscana and they influence development and stress tolerance. To further investigate J-domain protein function, ArHsp40 and ArHsp40-2 were each knocked down by RNA interference. Reductions in ArHsp40 and ArHsp40-2 had no effect on adult survival, time to release of cysts and nauplii from females and first-brood size. However, knockdown of both A. franciscana J-domain proteins reduced the longevity and heat tolerance of nauplii, with the loss of ArHsp40 having a greater effect. The knockdown of ArHsp40, but not of ArHsp40-2, caused approximately 50% of cysts to abort diapause entry and hatch without exposure to an exogenous signal such as low temperature and/or desiccation. Cysts lacking ArHsp40 that entered diapause exhibited decreased stress tolerance as did cysts with reduced ArHsp40-2, the latter to a lesser degree. The longevity of nauplii hatching prematurely from cysts was less than for nauplii arising by other means. The results expand our understanding of Hsp40 function in A. franciscana stress tolerance and development, especially during diapause, and they provide the first example of a molecular chaperone that influences diapause entry., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

World production of plastic has dramatically increased from the 1950's and now it reaches approximately 311 million tons per year. The resulting accumulation of small plastic detritus less than 5 mm in size, termed "microplastics", has started threatening the life cycles of marine organisms. Here we show the first evidence that microplastics disturb the initiation of symbiotic relationships in anthozoan-algae symbiosis. We found in both the aposymbiotic sea-anemone Aiptasia sp. and the coral Favites chinensis that the infectivity of symbiotic algae into the host is severely suppressed by microspheres fed either directly or indirectly through microsphere-fed Artemia sp. Similar trends were seen when microplastics collected from commercial facewash were used instead of microspheres. Therefore, ongoing accumulation of microplastics in the ocean might disturb the healthy anthozoan-algae symbiotic relationships, which are cornerstones of the biologically enriched coral reef ecosystem., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Marine sponges and cyanobacteria have a long history of co-evolution, with documented genome adaptations in cyanobionts. Both organisms are known to produce a wide variety of natural compounds, with only scarce information about novel natural compounds produced by cyanobionts. In the present study, we aimed to address their toxicological potential, isolating cyanobacteria ( n = 12) from different sponge species from the coast of Portugal (mainland, Azores, and Madeira Islands). After large-scale growth, we obtained both organic and aqueous extracts to perform a series of ecologically-relevant bioassays. In the acute toxicity assay, using nauplii of Artemia salina , only organic extracts showed lethality, especially in picocyanobacterial strains. In the bioassay with Paracentrotus lividus , both organic and aqueous extracts produced embryogenic toxicity (respectively 58% and 36%), pointing to the presence of compounds that interfere with growth factors on cells. No development of pluteus larvae was observed for the organic extract of the strain Chroococcales 6MA13ti, indicating the presence of compounds that affect skeleton formation. In the hemolytic assay, none of the extracts induced red blood cells lysis. Organic extracts, especially from picoplanktonic strains, proved to be the most promising for future bioassay-guided fractionation and compounds isolation. This approach allows us to classify the compounds extracted from the cyanobacteria into effect categories and bioactivity profiles.

Organisms increasingly encounter higher frequencies of extreme weather events as a consequence of global climate change. Currently, few strategies are available to mitigate climate change effects on animals arising from acute extreme high-temperature events. We tested the capacity of physiological engineering to influence the intra- and multi-generational upper thermal tolerance capacity of a model organism, Artemia , subjected to extreme high temperatures. Enhancement of specific physiological regulators during development could affect thermal tolerance or life-history attributes affecting subsequent fitness. Using experimental Artemia populations, we exposed F0 individuals to one of four treatments: heat hardening (28°C to 36°C, 1°C per 10 min), heat hardening plus serotonin (0.056 μg ml -1 ), heat hardening plus methionine (0.79 mg ml -1 ) and a control treatment. Regulator concentrations were based on previous literature. Serotonin may promote thermal tolerance, acting upon metabolism and life history. Methionine acts as a methylation agent across generations. For all groups, measurements were collected for three performance traits of individual thermal tolerance (upper sublethal thermal limit, lethal limit and dysregulation range) over two generations. The results showed that no treatment increased the upper thermal limit during acute thermal stress, although serotonin-treated and methionine-treated individuals outperformed controls across multiple thermal performance traits. Additionally, some effects were evident across generations. Together, these results suggest that phenotypic engineering provides complex outcomes, and if implemented with heat hardening can further influence performance in multiple thermal tolerance traits, within and across generations. Potentially, such techniques could be up-scaled to provide resilience and stability in populations susceptible to extreme temperature events., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

The current body of work on rearing larval/juvenile zebrafish is based on (1) utilization of freshwater and (2) diurnal light/dark cycle, (3) provision of live feed at modest density, and (4) culture in high visibility environment. We challenged these rearing approaches by maintaining zebrafish under constant light for 46-48 days (days postfertilization [dpf]), while securing continuous feeding in high turbidity and saline (1.8-2.1 parts per thousand) environment for the experiment's duration, allowing 24 h feeding/growth of fish from first exogenous feeding to maturation. There was no evidence of negative effects on zebrafish larvae behavior, growth, survival, and life cycle duration at constant illumination when food was continuously available. Zebrafish were stocked at high initial density (100 larvae/L) in a static system and fed high densities of rotifers (Brachionus plicatilis) (200-400/mL) from 6 to 12 dpf. Fish density was then reduced by 50% and two diet treatments, live rotifers or brine shrimp (Artemia) nauplii (10/mL), followed. Fish were reared on these two diets until first maturation. Performance of adult zebrafish fed live rotifer followed by Artemia nauplii diet was the highest recorded in the literature after 42 dpf, 250 ± 29 (males) and 430 ± 5 mg (females). Use of these rearing conditions, during the entire life cycle, until reproduction, resulted in the shortest ever recorded generation time (from egg to egg) of 43-45 dpf and fertilization rate (1 dpf) of 80.3%-94%.

Bio-active synthetic fabrics based on polyester (PET) and Nylon were manufactured by in-situ formation of Cu-BTC metal organic framework (MOF). In-growth of Cu-BTC within fabrics was accomplished in one pot simple process. The scanning microscope, X-ray diffraction and infrared spectra were all confirmed the formation of Cu-BTC within fabrics structure and reflected the role of fabrics' building unit in the Cu-BTC preparation. The estimated contents of materials onto fabrics were ranged in 97.14-127.33 mg MOF/g fabric and 30.59-40.10 mg Cu/g fabric. After embracing with Cu-BTC, color of fabrics was transformed to greenish-blue. The so-produced Cu-BTC/fabric hybrids were exhibited good biological activities against three different microbial pathogens (E. coli, S. aureus and C. albicans). The minimal inhibitory concentrations from the residual Cu-BTC powder were 65-70, 60-64 and 62-67 mg/mL, for S. aureus, E. coli and C. albicans pathogens, respectively, which were similar to that reported for commercial Cu-BTC. Moreover, no toxicity was observably detected for the released Cu-BTC from fabrics against brine shrimp at 10 mg/mL. These results revealed that, the in-growth of Cu-BTC resulted in production of biocidal synthetic fabrics without any ecotoxic effects at the as-used Cu-BTC content., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Biologically generated turbulence has been proposed as an important contributor to nutrient transport and ocean mixing 1-3 . However, to produce non-negligible transport and mixing, such turbulence must produce eddies at scales comparable to the length scales of stratification in the ocean. It has previously been argued that biologically generated turbulence is limited to the scale of the individual animals involved 4 , which would make turbulence created by highly abundant centimetre-scale zooplankton such as krill irrelevant to ocean mixing. Their small size notwithstanding, zooplankton form dense aggregations tens of metres in vertical extent as they undergo diurnal vertical migration over hundreds of metres 3,5,6 . This behaviour potentially introduces additional length scales-such as the scale of the aggregation-that are of relevance to animal interactions with the surrounding water column. Here we show that the collective vertical migration of centimetre-scale swimmers-as represented by the brine shrimp Artemia salina-generates aggregation-scale eddies that mix a stable density stratification, resulting in an effective turbulent diffusivity up to three orders of magnitude larger than the molecular diffusivity of salt. These observed large-scale mixing eddies are the result of flow in the wakes of the individual organisms coalescing to form a large-scale downward jet during upward swimming, even in the presence of a strong density stratification relative to typical values observed in the ocean. The results illustrate the potential for marine zooplankton to considerably alter the physical and biogeochemical structure of the water column, with potentially widespread effects owing to their high abundance in climatically important regions of the ocean 7 .

Many plants and animals store toxic or unpalatable compounds in tissues that are easily encountered by predators during attack. Defensive compounds can be produced de novo, or obtained from dietary sources and stored directly without selection or modification, or can be selectively sequestered or biotransformed. Storage strategies should be optimized to produce effective defence mechanisms but also prevent autotoxicity of the host. Nudibranch molluscs utilize a diverse range of chemical defences, and we investigated the accumulation and distribution of defensive secondary metabolites in body tissues of 19 species of Chromodorididae nudibranchs. We report different patterns of distribution across tissues, where: 1) the mantle had more or different (but structurally related) compounds than the viscera; 2) all compounds in the mantle were also in the viscera; and 3) the mantle had fewer compounds than the viscera. We found no further examples of species that selectively store a single compound, previously reported in Chromodoris species. Consistent with other studies, we found high concentrations of metabolites in mantle rim tissues compared to the viscera. Using bioassays, compounds in the mantle were more toxic than compounds found in the viscera for Glossodoris vespa Rudman, 1990 and Ceratosoma brevicaudatum Abraham, 1876. In G. vespa, compounds in the mantle were also more unpalatable to palaemonid shrimp than compounds found in the viscera. This indicates that these species may modify compounds to increase bioactivity for defensive purposes and/or selectively store more toxic compounds. We highlight clear differences in the storage of sequestered chemical defences, which may have important implications for species to employ effective defences against a range of predators.

The gradual increase of CO 2 concentration in the atmosphere, absorbed by the ocean surface water through air to sea equilibration termed ocean acidification (OA), leads to the decline of pH in seawater. It is not clear so far how the composition of fatty acids, particular the immune-related, in marine crustacean and the subsequent energy supply in marine ecosystem are affected by OA. The brine shrimp Artemia sinica is an open and common feed that provide essential fatty acids for mariculture. In this study, the fatty acids profiles of brine shrimp cultured under different lower pH levels of CO 2 driven seawater were investigated. The results showed a significant reduction of the proportion of total saturated fatty acids under the pH7.6 within one week. Meanwhile, the percentage of total monounsaturated fatty acids was significantly decreased at day 14 under pH7.8, and this percentage gave a significant increase of proportion within one week under pH7.6. Furthermore, the relative content of total polyunsaturated fatty acids (PUFAs) was found to be clearly increased with exposure to different seawater acidification at day 1, suggesting that the brine shrimp immune response was likely to be affected by acidified seawater as the PUFAs have been well known to be involved in immunomodulatory effects through alterations on cell membrane fluidity/lipid mediators and gene expression of cell signaling pathways. Notably, eicosapentaenoic acid and docosahexaenoic acid, which have essential effect on various physiological processes such as inflammatory cytokines production and cell structural stability, were strongly increased under two lower pH treatments within one week and with the significant increase at day 1 under pH7.6. These data clearly supported the hypothesis that OA might affect fatty acids composition, likely also the innate immunity, in crustacean and the subsequent energy transfer by food-chain system in the marine ecosystem., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Microplastic pollution has been a growing concern in the aquatic environment for several years. The abundance of microplastics in the environment has invariably led them to interact with a variety of different aquatic species. The small size of microplastics may make them bioavailable to a great range of species however, the impact this may have is not fully understood. Much of the research on microplastic pollution has focused on the marine environment and species with little research undertaken in freshwater. Here we examine the effect of microplastics on the freshwater cnidarian, Hydra attenuata. This study also describes the development and use of a bioassay to investigate the impact of microplastic on freshwater organisms. Hydra attenuata play a vital role in the planktonic make up of slow moving freshwater bodies which they inhabit and are sensitive environmental indicators. Hydra attenuata were exposed to polyethylene flakes (<400 ìm) extracted from facewash at different concentrations (Control, 0.01, 0.02, 0.04, 0.08 g mL -1 ). The ecologically relevant endpoint of feeding was measured by determining the amount of prey consumed (Artemia salina) after 30 and 60 min. The amount of microplastics ingested was also recorded at 30 min and 60 min. After which Hydra attenuata were transferred to clean media and observed after 3, 24, 48 & 96 h with changes in their morphology and reproduction (Hydranth numbers) recorded. The results of this study show that Hydra attenuata are capable of ingesting microplastics, with several individuals completely filling their gastric cavities. Significant reductions in feeding rates were observed after 30 min in 0.02 & 0.08 g mL -1 and after 60 min in 0.04 & 0.08 g mL -1 exposures. Exposure to the microplastics caused significant changes to the morphology of Hydra attenuata, however these changes were non-lethal. This study demonstrates that freshwater Hydra attenuata is capable of ingesting microplastics and that microplastic can significantly impact the feeding of freshwater organisms., (Copyright © 2017. Published by Elsevier Ltd.)

Chemical cues from fish, or kairomones, often impact the behavior of zooplankton. These behavioral changes are thought to improve predator avoidance. For example, marine and estuarine crustacean zooplankton become more sensitive to light after kairomone exposure, which likely deepens their vertical distribution into darker waters during the day and thereby reduces their visibility to fish predators. Here, we show that kairomones from an estuarine fish induce similar behavioral responses in adult brine shrimp (Artemia franciscana) from an endorheic, hypersaline lake, Great Salt Lake, Utah, USA. Given downwelling light stimuli, kairomone-exposed A. franciscana induce a descent response upon dimmer light flashes than they do in the absence of kairomones. Using extracellular electroretinogram (ERG) recordings, we also find that kairomones induce physiological changes in the retina that may lead to increased visual sensitivity, suggesting that kairomone-induced changes to photobehavior are mediated at the photoreceptor level. However, kairomones did not induce structural changes within the eye. Although A. franciscana inhabit endorheic environments that are too saline for most fish, kairomones from an estuarine fish amplify photobehavior in these branchiopod crustaceans. The mechanism for this behavioral change has both similarities to and differences from that described in marine malacostracan crustaceans.

In most sexual, diploid eukaryotes, at least one crossover occurs between each pair of homologous chromosomes during meiosis, presumably in order to ensure proper segregation. Well-known exceptions to this rule are species in which one sex does not recombine and specific chromosomes lacking crossover. We review other possible exceptions, including species with chromosome maps of less than 50 cM in one or both sexes. We discuss the idea that low recombination rates may favour sex-asex transitions, or, alternatively may be a consequence of it. We then show that a yet undescribed species of brine shrimp Artemia from Kazakhstan ( A sp. Kazakhstan), the closest known relative of the asexual Artemia parthenogenetica , has one of the shortest genetic linkage maps known. Based on a family of 42 individuals and 589 RAD markers, we find that many linkage groups are considerably shorter than 50 cM, suggesting either no obligate crossover or crossovers concentrated at terminal positions with little effect on recombination. We contrast these findings with the published map of the more distantly related sexual congener, A. franciscana , and conclude that the study of recombination in non-model systems is important to understand the evolutionary causes and consequences of recombination.This article is part of the themed issue 'Evolutionary causes and consequences of recombination rate variation in sexual organisms'., (© 2017 The Author(s).)

The hydrothermal carbonization of sewage sludge has been studied as an alternative technique for the conversion of sewage sludge into value-added products, such as soil amendments. We tested the toxicity of biosolid hydrochar (Sewchar) to earthworms. Additionally, the toxicity of Sewchar process water filtrate with and without pH adjustment was assessed, using brine shrimps as a model organism. For a Sewchar application of 40 Mg ha -1 , the earthworms significantly preferred the side of the vessel with the reference soil (control) over side of the vessel with the Sewchar treatments. There was no acute toxicity of Sewchar to earthworms within the studied concentration range (up to 80 Mg ha -1 ). Regarding the Sewchar process water filtrate, the median lethal concentration (LC50) to the shrimps was 8.1% for the treatments in which the pH was not adjusted and 54.8% for the treatments in which the pH was adjusted to 8.5. The lethality to the shrimps significantly increased as the amount of Sewchar process water filtrate increased. In the future, specific toxic substances in Sewchar and its process water filtrate, as well as their interactions with soil properties and their impacts on organisms, should be elucidated. Additionally, it should be identified whether the amount of the toxic compounds satisfies the corresponding legal requirements for the safe application of Sewchar and its process water filtrate.

Bacterial infections are major constraints in aquaculture farming. These pathogens often adapt to the biofilm mode of growth and resist antibiotic treatments. We have used a non-toxic glycolipid biosurfactant (BS-SLSZ2) derived from a marine epizootic bacterium Staphylococcus lentus to treat aquaculture associated infections in an eco-friendly manner. We found that BS-SLSZ2 contained threose, a four-carbon sugar as the glycone component, and hexadecanoic and octadecanoic acids as the aglycone components. The critical micelle concentration of the purified glycolipid was 18 mg mL-1. This biosurfactant displayed anti-adhesive activity and inhibited biofilm formation by preventing initial attachment of cells onto surfaces. The biosurfactant (at a concentration of 20 μg) was able to inhibit Vibrio harveyi and Pseudomonas aeruginosa biofilms by 80.33 ± 2.16 and 82 ± 2.03%, respectively. At this concentration, it was also able to disrupt mature biofilms of V. harveyi (78.7 ± 1.93%) and P. aeruginosa (81.7 ± 0.59%). The biosurfactant was non-toxic towards Artemia salina. In vivo challenge experiments showed that the glycolipid was effective in protecting A. salina nauplii against V. harveyi and P. aeruginosa infections. This study highlights the significance of marine natural products in providing alternative biofilm controlling agents and decreasing the usage of antibiotics in aquaculture settings., (© FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Using Artemia salina (A. salina) cysts (capsulated and decapsulated) and larvae [instar I (0-24 h), II (24-48 h) and III (48-72 h)] as experimental models, developmental toxicity of oxidized multi-walled carbon nanotubes (O-MWCNTs) was evaluated. Results revealed that hatchability of capsulated and decapsulated cysts was significantly decreased (p < 0.01) following exposure to 600 mg/L for 36 h. Mortality rates were 33.8, 55.7 and 40.7% for instar I, II and III larvae in 600 mg/L. The EC 50 values for swimming inhibition of instar I, II and III were 535, 385 and 472 mg/L, respectively. Instar II showed the greatest sensitivity to O-MWCNTs, and followed by instar III, instar I, decapsulated cysts and capsulated cysts. Effects on hatchability, mortality and swimming were accounted for O-MWCNTs rather than metal catalyst impurities. Body length was decreased with the concentrations increased from 0 to 600 mg/L. O-MWCNTs attached onto the cysts, gill and body surface, resulting in irreversible damages. Reactive oxygen species, malondialdehyde content, total antioxidant capacity and antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) activities were increased following exposure, indicating that the effects were related to oxidative stress. O-MWCNTs were ingested and distributed in phagocyte, lipid vesicle and intestine. Most of the accumulated O-MWCNTs were excreted by A. salina at 72 h, but some still remained in the organism. Data of uptake kinetics showed that O-MWCNTs contents in A. salina were gradually increased from 1 to 48 h and followed by rapidly decreased from 48 to 72 h with a range from 5.5 to 28.1 mg/g. These results so far indicate that O-MWCNTs have the potential to affect aquatic organisms when released into the marine ecosystems., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The populations of Artemia (or brine shrimp) from the Americas exhibit a wide variation in the amount of interphase heterochromatin. There is interest in understanding how this variation affects different parameters, from the cellular to the organismal levels. This should help to clarify the ability of this organism to tolerate brine habitats regularly subject to strong abiotic changes. In this study, we assessed the amount of interphase heterochromatin per nucleus based on chromocenter number (N-CHR) and relative area of chromocenter (R-CHR) in two species of Artemia, A. franciscana (Kellog, 1906) (n=9 populations) and A. persimilis (Piccinelli and Prosdocimi, 1968) (n=3 populations), to investigate the effect on nuclear size (S-NUC). The relationship of the R-CHR parameter with the ionic composition (IC) of brine habitats was also analysed. Our results indicate a significant variation in the amount of heterochromatin both within and between species (ANOVA, p<0.001). The heterochromatin varied from 0.81 ± 1.17 to 12.58 ± 3.78 and from 0.19 ± 0.34% to 11.78 ± 3.71% across all populations, for N-CHR and R-CHR parameters, respectively. N-CHR showed less variation than R-CHR (variation index 15.5-fold vs. 62-fold). At least five populations showed a significant association (p<0.05) between R-CHR and S-NUC, either with negative (four populations, r= from -0.643 to -0.443), or positive (one population, r= 0.367) values.Within each species, there were no significant associations between both parameters (p>0.05). The R-CHR and IC parameters were associated significantly for the magnesium ion (r= 0.496, p<0.05) and also for the chloride, sodium and calcium ions (r = from -0.705 to -0.478, p<0.05). At species level, a significant association between both parameters was also found in A. franciscana populations, for the sulphate and calcium ions, in contrast to A. persimilis. These findings suggest that the amount of interphase heterochromatin modifies the nuclear size in Artemia. Our data also indicate that change in the amount of interphase heterochromatin is in line with the ionic composition of brines. This would be a species-specific phenomenon, whose occurrence may be involved in the ability of this organism to survive in these environments.

The present study deals with the toxicity assessment of two differently synthesized zero valent iron nanoparticles (nZVI, chemical and biological) as well as Fe 2+ ions on Artemia salina at three different initial concentrations of 1, 10, and 100 mg/L of these particles. The assessment was done till 96 h at time intervals of 24 h. EC 50 value was calculated to evaluate the 50% mortality of Artemia salina at all exposure time durations. Between chemically and biologically synthesized nZVI nanoparticles, insignificant differences in the level of mortality were demonstrated. At even 24 h, Fe 2+ ion imparted complete lethality at the highest exposure concentration (100 mg/L). To understand intracellular oxidative stress because of zero valent iron nanoparticles, ROS estimation, SOD activity, GSH activity, and catalase activity was performed which demonstrated that ionic form of iron is quite lethal at high concentrations as compared with the same concentration of nZVI exposure. Lower concentrations of nZVI were more toxic as compared with the ionic form and was in order of CS-nZVI > BS-nZVI > Fe 2+ . Cell membrane damage and bio-uptake of nanoparticles were also evaluated for all three concentrations of BS-nZVI, CS-nZVI, and Fe 2+ using adult Artemia salina in marine water; both of which supported the observations made in toxicity assessment. This study can be further explored to exploit Artemia salina as a model organism and a biomarker in an nZVI prone aquatic system to detect toxic levels of these nanoparticles. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1617-1627, 2017., (© 2017 Wiley Periodicals, Inc.)

Harmful algae may differently affect their primary grazers, causing sub-lethal effects and/or leading to their death. The present study aim to compare the effects of three toxic benthic dinoflagellates on clearance and grazing rates, behavioral changes, and survival of Artemia salina. Feeding assays consisted in 1-h incubations of brine shrimps with the toxic Prorocentrum lima, Gambierdiscus excentricus and Ostreopsis cf. ovata and the non-toxic Tetraselmis sp. Brine shrimps fed unselectively on all toxic and non-toxic algal preys, without significant differences in clearance and ingestion rates. Acute toxicity assays were performed with dinoflagellate cells in two growth phases during 7-h to assess differences in cell toxicity to A. salina. Additionally, exposure to cell-free medium was performed to evaluate its effects on A. salina survival. The behavior of brine shrimps significantly changed during exposure to the toxic dinoflagellates, becoming immobile at the bottom by the end of the trials. Dinoflagellates significantly affected A. salina survival with 100% mortality after 7-h exposure to cells in exponential phase (all treatments) and to P. lima in stationary phase. Mortality rates of brine shrimps exposed to O. cf. ovata and G. excentricus in stationary phase were 91% and 75%, respectively. However, incubations of the brine shrimps with cell-free medium did not affect A. salina survivorship. Significant differences in toxic effects between cell growth phases were only found in the survival rates of A. salina exposed to G. excentricus. Acute exposure to benthic toxic dinoflagellates induced harmful effects on behavior and survival of A. salina. Negative effects related to the toxicity of benthic dinoflagellates are thus expected on their primary grazers making them more vulnerable to predation and vectors of toxins through the marine food webs.

An aerobic marine bacterium Vibrio sp. Y1-5 was screened to achieve efficient nitrate and ammonium removal simultaneously and fix nitrogen in cells without N loss. Approximately 98.0% of nitrate (100mg/L) was removed in 48h through assimilatory nitrate reduction and nitrate reductase was detected in the cytoplasm. Instead of nitrification, the strain assimilated ammonium directly, and it could tolerate as high as 1600mg/L ammonium concentration while removing 844.6mg/L. In addition, ammonium assimilation occurred preferentially in the medium containing nitrate and ammonium with a total nitrogen (TN) removal efficiency of 80.4%. The results of nitrogen balance and Fourier infrared spectra illustrated that the removed nitrogen was all transformed to protein or stored as organic nitrogen substances in cells and no N was lost in the process. Toxicological studies with the brine shrimp species Artemia naupliia indicated that Vibrio sp. Y1-5 can be applied in aquatic ecosystems safely., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Nanosized titanium dioxide (nTiO 2 ) is widespread in many commercial products and several authors investigated its ecotoxicity effects focusing mainly on freshwater environments. Data on saltwater species are still lacking or present contradicting results. We compared for the first time the toxicity of TiCl 4 and nTiO 2 considering standard toxicity tests with microalgae Phaeodactylum tricornutum (growth inhibition test, 1.8-90mg/L) and crustacean Artemia franciscana (mortality test, 0.5-64mg/L). For A. franciscana, two alternative scenarios were considered beside standard protocol: i) darkness; and ii) starvation. About microalgae, results evidenced that effects of TiCl 4 (EC50=63mg/L) were greater than nTiO 2 (no EC50), but IC10 and IC20 were significantly lower suggesting that nTiO 2 is more harmful than TiCl 4 at lower concentrations. The effects of TiCl 4 to crustaceans larvae in all exposure scenarios were lower compared to nTiO 2 (EC50(96h)=15mg/L - standard protocol). During toxicity testing, the absence of light generally lowered nTiO 2 effects while starvation increased the toxicity of both TiCl 4 and nTiO 2 ., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Gradually increasing atmospheric CO 2 partial pressure (pCO 2 ) has caused an imbalance in carbonate chemistry and resulted in decreased seawater pH in marine ecosystems, termed seawater acidification. Anthropogenic seawater acidification is postulated to affect the physiology of many marine calcifying organisms. To understand the possible effects of seawater acidification on the proteomic responses of a marine crustacean brine shrimp (Artemia sinica) three groups of cysts were hatched and further raised in seawater at different pH levels (8.2 as control and 7.8 and 7.6 as acidification stress levels according to the predicted levels at the end of this century and next century, respectively) for 1, 7 and 14 days followed by examination of the protein expression changes via two-dimensional gel electrophoresis. Searches of protein databases revealed that 67 differential protein spots were altered due to lower pH level (7.6 and 7.8) stress in comparison to control groups (pH 8.2) by mass spectrometry. Generally, these differentially expressed proteins included the following: 1) metabolic process-related proteins involved in glycolysis and glucogenesis, nucleotide/amino acid/fatty acid metabolism, protein biosynthesis, DNA replication and apoptosis; 2) stress response-related proteins, such as peroxiredoxin, thioredoxin peroxidase, 70-kDa heat shock protein, Na/K ATPase, and ubiquinol-cytochrome c reductase; 3) immune defence-related proteins, such as prophenoloxidase and ferritin; 4) cytoskeletal-related proteins, such as myosin light chain, TCP1 subunit 2, tropomyosin and tubulin alpha chain; and 5) signal transduction-related proteins, such as phospholipase C-like protein, 14-3-3 zeta, translationally controlled tumour protein and RNA binding motif protein. Taken together, these data support the idea that CO 2 -driven seawater acidification may affect protein expression in the crustacean A. sinica and possibly also in other species that feed on brine shrimp in the ecosystem, particularly marine food webs., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Modeling toxicity of metal mixtures poses unique challenges to the incorporation of bioavailability and metal speciation in metal exposures. Three models (models I, II, and III) were compared in the present study to predict and interpret the toxicity exerted by binary metal mixtures to zebrafish larvae, with the assumption of competition between metals based on the biotic ligand model and toxic potencies of individual metals. In addition, 3 models were developed by substituting binding constants (f MBL ) for internal metal concentrations (C M,int ) to directly delineate single-metal and mixture effects on mortality of zebrafish larvae. The results indicated that the 3 developed models appeared to be much better (p < 0.01) than 3 previous models at assessing the toxicity of different metal mixtures and showed 10% to 20% predictive improvement for each metal combination, with the toxic equivalency factor-based model II showing the best performance at quantifying metal mixture toxicity. The 3 developed models generally provided a reasonable framework and descriptions of bioavailability and additive (or nearly additive) toxicity for a number of binary metal mixtures. Environ Toxicol Chem 2016;35:2721-2733. © 2016 SETAC., (© 2016 SETAC.)

Stressful environments affect life-history components of fitness through (i) instantaneous detrimental effects, (ii) historical (carry-over) effects and (iii) history-by-environment interactions, including acclimation effects. The relative contributions of these different responses to environmental stress are likely to change along life, but such ontogenic perspective is often overlooked in studies of tolerance curves, precluding a better understanding of the causes of costs of acclimation, and more generally of fitness in temporally fine-grained environments. We performed an experiment in the brine shrimp Artemia to disentangle these different contributions to environmental tolerance, and investigate how they unfold along life. We placed individuals from three clones of A. parthenogenetica over a range of salinities during a week, before transferring them to a (possibly) different salinity for the rest of their lives. We monitored individual survival at repeated intervals throughout life, instead of measuring survival or performance at a given point in time, as commonly done in acclimation experiments. We then designed a modified survival analysis model to estimate phase-specific hazard rates, accounting for the fact that individuals may share the same treatment for only part of their lives. Our approach allowed us to distinguish effects of salinity on (i) instantaneous mortality in each phase (habitat quality effects), (ii) mortality later in life (history effects) and (iii) their interaction. We showed clear effects of early salinity on late survival and interactions between effects of past and current environments on survival. Importantly, analysis of the ontogenetic dynamics of the tolerance curve reveals that acclimation affects different parts of the curve at different ages. Adopting a dynamical view of the ontogeny of tolerance curve should prove useful for understanding niche limits in temporally changing environments, where the full sequence of environments experienced by an individual determines its overall environmental tolerance, and how it changes throughout life., (© 2016 The Authors. Journal of Animal Ecology © 2016 British Ecological Society.)

Filter-feeding organisms are often keystone species with a major influence on the dynamics of aquatic ecosystems. Studies of filtering rates in such taxa are therefore vital in order to understand ecosystem functioning and the impact of natural and anthropogenic stressors such as parasites, climate warming and invasive species. Brine shrimps Artemia spp. are the dominant grazers in hypersaline systems and are a good example of such keystone taxa. Hypersaline ecosystems are relatively simplified environments compared with much more complex freshwater and marine ecosystems, making them suitable model systems to address these questions. The aim of this study was to compare feeding rates at different salinities and temperatures between clonal A. parthenogenetica (native to Eurasia and Africa) and the invasive American brine shrimp A. franciscana, which is excluding native Artemia from many localities. We considered how differences observed in laboratory experiments upscale at the ecosystem level across both spatial and temporal scales (as indicated by chlorophyll-a concentration and turbidity). In laboratory experiments, feeding rates increased at higher temperatures and salinities in both Artemia species and sexes, whilst A. franciscana consistently fed at higher rates. A field study of temporal dynamics revealed significantly higher concentrations of chlorophyll-a in sites occupied by A. parthenogenetica, supporting our experimental findings. Artemia parthenogenetica density and biomass were negatively correlated with chlorophyll-a concentration at the spatial scale. We also tested the effect of cestode parasites, which are highly prevalent in native Artemia but much rarer in the invasive species. The cestodes Flamingolepis liguloides and Anomotaenia tringae decreased feeding rates in native Artemia, whilst Confluaria podicipina had no significant effect. Total parasite prevalence was positively correlated with turbidity. Overall, parasites are likely to reduce feeding rates in the field, and their negative impact on host fecundity is likely to exacerbate the difference between grazing rates of native and alien Artemia populations at the ecosystem level. The results of this study provide evidence for the first time that the replacement of native Artemia by A. franciscana may have major consequences for the functioning of hypersaline ecosystems. The strong effect of parasites on feeding rate underlines the importance of taking parasites into account in order to improve our understanding of the functioning of aquatic ecosystems., Competing Interests: The authors have declared that no competing interests exist.

Textile industry has led to severe environmental pollution and is posing a serious threat to the ecosystems. Immobilized biocatalysts have gained importance as potential bio-remediating agent. Manganese peroxidase (MnP) was immobilized onto glutaraldehyde activated chitosan beads by crosslinking and employed for the degradation and detoxification of dyes in textile effluents. The efficiency of chitosan-immobilized MnP (CI-MnP) was evaluated on the basis of decolorization, water quality improvement and toxicity reduction. Maximum color removal of 97.31% was recorded and up to 82.40%, 78.30% and 91.7% reductions in COD, TOC, and BOD were achieved, respectively. The cytotoxicity of bio-treated effluents reduced significantly and 38.46%, 43.47% and 41.83% Allium cepa root length, root count and mitotic index were increased, respectively, whereas brine shrimp nauplii death reduced up to 63.64%. Mutagenicity (Ames test) reduced up to 73.44% and 75.43% for TA98 and TA100 strains, respectively. The CI-MnP retained 60% activity after 10 repeated decolorization batches. The CI-MnP showed excellent efficiency for the bioremediation of textile effluents and can be used for the remediation of toxic agents in wastewater. The monitoring of processed wastewater using bioassays is suggested to evaluate bio-efficiency of treatment method for safe disposal of effluents into water bodies., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of this study was to evaluate the effects of feeding on synbiotic (Pediococcus acidilactici and fructooligosaccharide) enriched adult Artemia franciscana on skin mucus immune responses, stress resistance, intestinal microbiota and growth performance of angelfish (Pterophyllum scalare). Three hundred and sixty fish with initial weight 3.2 ± 0.13 g were randomly divided into twelve aquaria (50 L) assigned to four groups in triplicates. Fish were fed for 7 weeks with dietary treatments, including treatment 1: feeding adult Artemia without enrichment (control group), treatment 2: feeding adult Artemia enriched with lyophilised probiotic P. acidilactici (700 mg L(-1)), 3: feeding adult Artemia enriched with prebiotic fructooligosaccharide (FOS) (100 mg L(-1)), group 4: feeding adult Artemia enriched with synbiotic (P. acidilactici (700 mg L(-1)) + FOS (100 mg L(-1))). Skin mucus immune responses (lysozyme activity, total Immunoglobulin and protease), stress resistance against environmental stress (acute decrease of temperature and increase salinity), intestinal microbiota as well as growth indices were measured at the end of feeding trial. Artemia enriched with synbiotic significantly improved growth performance compared to other treatments (P < 0.05). The highest weight gain and specific growth rate (SGR) was observed in synbiotic fed fish (P < 0.05). Compared to the other treatments, the population of lactic acid bacteria was significantly higher in the intestinal microbiota of fish fed synbiotic supplemented diet (P < 0.05). In the environmental stress challenge test, the maximum resistance to abrupt decrease of temperature (17 °C) or elevation of salinity (12 g per liter) was observed in the synbiotic treatment. Also, the total immunoglobulin and lysozyme activity level of skin mucus was significantly elevated in fish fed Artemia enriched with synbiotic (P < 0.05). These results revealed that feeding angelfish with synbiotic enriched Artemia was more effective than singular enrichment with probiotics or prebiotics., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Sex allocation theory is often hailed as the most successful area of evolutionary theory due to its striking success as a predictor of empirical observations [1]. Most naturally occurring sex ratios can be explained by the principle of equal investment in the sexes [2-4] or by cases of "extraordinary" sex allocation [5]. Deviations from the expected sex ratio are often correlated with weak selection or low environmental predictability (e.g., [6, 7]); true cases of aberrant sex allocation are surprisingly rare [8]. Here, we present a case of long-lasting maladaptive sex allocation, which we discovered in invasive populations of the exclusively sexual brine shrimp Artemia franciscana. A. franciscana was introduced to Southern France roughly 500 generations ago [9]; since then, it has coexisted with the native asexual species Artemia parthenogenetica [10]. Although we expect A. franciscana to produce balanced offspring sex ratios, we regularly observed extremely male-biased sex ratios in invasive A. franciscana, which were significantly correlated to the proportion of asexuals in the overall population. We experimentally proved that both invasive- and native-range A. franciscana overproduced sons when exposed to excess females, without distinguishing between conspecific and asexual females. We conclude that A. franciscana adjust their offspring sex ratio in function of the adult sex ratio but are information limited in the presence of asexual females. Their facultative adjustment trait, which is presumably adaptive in their native range, has thus become maladaptive in the invasive range where asexuals occur. Despite this, it has persisted unchanged for hundreds of generations., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Cohort data are frequently collected to study stage-structured development and mortalities of many organisms, particularly arthropods. Such data can provide information on mean stage durations, among-individual variation in stage durations, and on mortality rates. Current statistical methods for cohort data lack flexibility in the specification of stage duration distributions and mortality rates. In this paper, we present a new method for fitting models of stage-duration distributions and mortality to cohort data. The method is based on a Monte Carlo within MCMC algorithm and provides Bayesian estimates of parameters of stage-structured cohort models. The algorithm is computationally demanding but allows for flexible specifications of stage-duration distributions and mortality rates. We illustrate the algorithm with an application to data from a previously published experiment on the development of brine shrimp from Mono Lake, California, through nine successive stages. In the experiment, three different food supply and temperature combination treatments were studied. We compare the mean duration of the stages among the treatments while simultaneously estimating mortality rates and among-individual variance of stage durations. The method promises to enable more detailed studies of development of both natural and experimental cohorts. An R package implementing the method and which allows flexible specification of stage duration distributions is provided.

Parasites and pollutants can both affect any living organism, and their interactions can be very important. To date, repeated studies have found that parasites and heavy metals or metalloids both have important negative effects on the health of animals, often in a synergistic manner. Here, we show for the first time that parasites can increase host resistance to metalloid arsenic, focusing on a clonal population of brine shrimp from the contaminated Odiel and Tinto estuary in SW Spain. We studied the effect of cestodes on the response of Artemia to arsenic (acute toxicity tests, 24h LC50) and found that infection consistently reduced mortality across a range of arsenic concentrations. An increase from 25°C to 29°C, simulating the change in mean temperature expected under climate change, increased arsenic toxicity, but the benefits of infection persisted. Infected individuals showed higher levels of catalase and glutathione reductase activity, antioxidant enzymes with a very important role in the protection against oxidative stress. Levels of TBARS were unaffected by parasites, suggesting that infection is not associated with oxidative damage. Moreover, infected Artemia had a higher number of carotenoid-rich lipid droplets which may also protect the host through the "survival of the fattest" principle and the antioxidant potential of carotenoids. This study illustrates the need to consider the multi-stress context (contaminants and temperature increase) in which host-parasite interactions occur.

Selective harvesting can cause evolutionary responses in populations via shifts in phenotypic characteristics, especially those affecting life history. Brine shrimp (Artemia franciscana) cysts in Great Salt Lake (GSL), Utah, USA are commercially harvested with techniques that select against floating cysts. This selective pressure could cause evolutionary changes over time. Our objectives are to (1) determine if there is a genetic basis to cyst buoyancy, (2) determine if cyst buoyancy and nauplii mortality have changed over time, and (3) to examine GSL environmental conditions over time to distinguish whether selective harvesting pressure or a trend in environmental conditions caused changes in cyst buoyancy and nauplii mortality. Mating crosses between floating and sinking parental phenotypes with two food concentrations (low and high) indicated there is a genetic basis to cyst buoyancy. Using cysts harvested from 1991-2011, we found cyst buoyancy decreased and nauplii mortality increased over time. Data on water temperature, salinity, and chlorophyll a concentration in GSL from 1994 to 2011 indicated that although water temperature has increased over time and chlorophyll a concentration has decreased over time, the selective harvesting pressure against floating cysts is a better predictor of changes in cyst buoyancy and nauplii mortality over time than trends in environmental conditions. Harvesting of GSL A. franciscana cysts is causing evolutionary changes, which has implications for the sustainable management and harvesting of these cysts. Monitoring phenotypic characteristics and life-history traits of the population should be implemented and appropriate responses taken to reduce the impacts of the selective harvesting.

When prey are scarce, planktivorous fish and other predators feeding on tiny prey should forage within prey-rich patches to attain a net food intake above the ambient mean food concentrations. If they can indeed locate prey-rich patches efficiently, then a patchy distribution of planktonic prey should lead to: (1) an increase in the overall per capita food intake, and (2) greater variability among predators in prey-capture rate due to differences in arrival times. Both phenomena were observed in 34 daily feeding sessions with a cohort of juvenile rudd held in twin experimental systems, each housing the same number of fish free to move in a loop of ten interconnected 200-L tanks. The fish were fed daily with equal numbers of planktonic prey (Artemia nauplii), offered either in a homogeneous or patchy distribution. To simulate low and high temperatures that represent potential global warming scenarios, the feeding protocol was replicated at 16, 21 and 26 °C, on each occasion following a 3-day period of fish acclimation. Up to 40-70 % of fish in the system with the patchy prey distribution assembled rapidly in the high-prey-density tank, the capture rate of first arrivals being up to 60 prey min(-1) at 26 °C, orders of magnitude greater than that of latecomers. The overall capture rates were higher in the system with patchy prey, regardless of the temperature. At the highest temperature (26 °C), the fish located the high-prey-density tank in less than half the time taken at the lowest temperature (16 °C, Q(10) > 2).