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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.)

Among natural products, essential oils from aromatic plants have been reported to possess potent anticancer properties. In this work, we aimed to perform the cytotoxic concentration range screening and antiproliferative activity screening of chemically characterized Thymus vulgaris L. essential oil. In vivo bioassay was conducted using the brine shrimp lethality test (BSLT). In vitro evaluation of antiproliferative activity was carried out on three human tumor cell lines: breast adenocarcinoma MCF-7, lung carcinoma H460 and acute lymphoblastic leukemia MOLT-4 using MTT assay. Essential oil components thymol (36.7%), p-cymene (30.0%), γ-terpinene (9.0%) and carvacrol (3.6%) were identified by gas chromatography/mass spectrometry. Analyzed essential oil should be considered as toxic/highly toxic with LC 50 60.38 µg/mL in BSLT and moderate/weakly cytotoxic with IC 50 range 52.65-228.78 µg/mL in vitro, according to evaluated cytotoxic criteria. Essential oil induced a dose-dependent inhibition of cell proliferation in all tested tumor cell lines and showed different sensitivity. Dose dependent toxicity observed in bioassay as well as the in vitro assay confirmed that brine shrimp lethality test is an adequate method for preliminary toxicity testing of Thymus vulgaris L. essential oil in tumor cell lines.

Background: Clausena excavata Burum. f. has long been applied in ethnomedicine for the treatment of various disorders like rhinitis, headache, cough, wound healing, fever, and detoxification. This study is aimed at investigating the antibacterial activity against Enterococcus faecalis ATCC 49532 using AlamarBlue assay and atomic force microscopy (AFM) as well as the cytotoxicity, anticancer, and phytotoxicity of C . excavata ., Method: Bacterial cell viability was performed by using microplate AlamarBlue assay. Atomic force microscopy was used to determine morphological changes in the surface of bacterial cells. Cytotoxicity and phytotoxicity were determined by brine shrimp lethality and Lemna minor bioassay. Caco-2 (colorectal adenocarcinoma) cell line was used for the evaluation of the anticancer effects., Result: Among the fractions tested, ethyl acetate (EA) fraction was found to be active with minimum inhibitory concentration (MIC) of 750  μ g/mL against E . faecalis , but other fractions were found to be insensitive to bacterial growth. Microscopically, the EA fraction-treated bacteria showed highly damaged cells with their cytoplasmic content scattered all over. The LC 50 value of the EA fraction against brine shrimp was more than 1000  μ g/mL showing the nontoxic nature of this fraction. Chloroform (CH), EA, and methanol (MOH) fractions of C . excavata were highly herbicidal at the concentration of 1000  μ g/mL. EA inhibited Caco-2 cell line with an IC 50 of 20  μ g/mL., Conclusions: This study is the first to reveal anti- E . faecalis property of EA fraction of C . excavata leaves, natural herbicidal, and anticancer agents thus highlight the potential compound present in its leaf which needs to be isolated and tested against multidrug-resistant E . faecalis ., Competing Interests: We wish to confirm that there are no known conflicts of interest associated with this publication., (Copyright © 2021 Shaymaa Fadhel Abbas Albaayit et al.)

Nine new xanthone-type and anthraquinone-type mycotoxins including austocystins J-N (1-5), 7-chloro versicolorin A (6), 3'-hydroxy-8-O-methyl versicolorin B (7), 8-O-methyl versiconol (8) and 2',3'-dihydroxy versiconol (9), together with 17 known analogues (10-26) were isolated from an extract of the deep-sea-derived fungus Aspergillus puniceus SCSIO z021. Their structures were elucidated by detailed analysis of spectroscopic data, and their absolute configurations were further determined by quantum chemical calculations of ECD spectra or comparison of the experimental ECD spectra. Eleven hydrogenated austocystins were synthesized from 1-2, 10-15 and 17 by catalytic hydrogenation for bioactivities evaluation. Totally, 18 of the all 37 compounds showed strong toxicity against brine shrimps or Vero cell, and the toxicity of 8-O-methyldemethylsterigmatocystin (18) (LC 50  = 0.020 µM) against brine shrimps was higher than those of three positive controls. In addition, 22 of the isolated compounds also exhibited significant inhibitory activity against seven different protein tyrosine phosphatases (PTPs), among them austocystin H (15) and methyl-averantin (24) were the most potent inhibitors with IC 50 values of 0.20-3.0 µM. Their structure-bioactivity relationship was also discussed., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Flowers-like ZnO structures were synthesized using Arabic Gum (AGZnO) or Karaya Gum (KGZnO). The AGZnO and KGZnO were characterized by X-ray diffractometry, Fourier Transformed Infrared, Scanning Electron Microscopy, Photoluminescence, nitrogen adsorption/desorption and diffuse reflectance techniques. The materials were tested in the discoloration of Methylene Blue (MB) dye under visible light and scavenger studies were also performed. The toxicity of the MB irradiated was investigated in bioassays with Artemia salina. The structural characterization demonstrated the formation of hexagonal ZnO. All samples presented flower-like morphology with presence of mesopores identified by BET method. The optical properties indicated band gap of 2.99 (AGZnO) and 2.76 eV (KGZnO), and emission in violet, blue and green emissions also were observed. The KGZnO demonstrated better photocatalytic performance than the AGZnO, and scavenger studies indicated that OH radicals are the main species involved in the degradation of the pollutant model. The photodiscoloration of MB solution did not demonstrate toxicity. Therefore, KGZnO is a promising material for photocatalysis application., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Callus, suspension and bioreactor cultures of Verbena officinalis were established, and optimized for biomass growth and production of phenylpropanoid glycosides, phenolic acids, flavonoids and iridoids. All types of cultures were maintained on/in the Murashige and Skoog (MS) media with 1 mg/L BAP and 1 mg/L NAA. The inoculum sizes were optimized in callus and suspension cultures. Moreover, the growth of the culture in two different types of bioreactors-a balloon bioreactor (BB) and a stirred-tank bioreactor (STB) was tested. In methanolic extracts from biomass of all types of in vitro cultures the presence of the same metabolites-verbascoside, isoverbascoside, and six phenolic acids: protocatechuic, chlorogenic, vanillic, caffeic, ferulic and rosmarinic acids was confirmed and quantified by the HPLC-DAD method. In the extracts from lyophilized culture media, no metabolites were found. The main metabolites in biomass extracts were verbascoside and isoverbascoside. Their maximum amounts in g/100 g DW (dry weight) in the tested types of cultures were as follow: 7.25 and 0.61 (callus), 7.06 and 0.48 (suspension), 7.69 and 0.31 (BB), 9.18 and 0.34 (STB). The amounts of phenolic acids were many times lower, max. total content reached of 26.90, 50.72, 19.88, and 36.78 mg/100 g DW, respectively. The highest content of verbascoside and also a high content of isoverbascoside obtained in STB (stirred-tank bioreactor) were 5.3 and 7.8 times higher than in extracts from overground parts of the parent plant. In the extracts from parent plant two iridoids-verbenalin and hastatoside, were also abundant. All investigated biomass extracts and the extracts from parent plant showed the antiproliferative, antioxidant and antibacterial activities. The strongest activities were documented for the cultures maintained in STB. We propose extracts from in vitro cultured biomass of vervain, especially from STB, as a rich source of bioactive metabolites with antiproliferative, antioxidant and antibacterial properties.

Skin darkening results as a consequence of the accumulation of skin pigment melanin. To combat this, the amplitude of skin lightening agents are commercially available, most of which inhibit melanin synthesis. Decolorization of melanin is an alternative method of skin lightening. In this study, we show that lignin peroxidase (LiP), an extracellular enzyme purified from Phanerochaete chrysosporium NK-1 isolated from a forest soil can effectively degrade and decolorize melanin in vitro. Decolorization conditions including pH, temperature, incubation time, enzyme concentration, and mediator addition were investigated to optimize the reaction conditions. The results indicate that pH 3, 40 °C, 15 IU/ml, and 10 h incubation were the optimal conditions for the decolorization of the melanin. The use of the mediator, veratryl alcohol was also found effective to enhance the efficacy of the melanin decolonization, with up to 92% decolorization. The scanning electron microscopy results showed void spaces on the treated melanin granules as compared to the untreated sample, indicating the degradation of melanin. Changes in the fingerprint region of the melanin were observed. Between wavenumbers 1500-500 cm -1 , for example, the presence of new peaks in the treated melanin at 1513, 1464, and 1139 cm -1 CH 2 , CH 3  bend and C-O-C stretch represented structural changes. A new peak at 2144 cm -1 (alkynyl C≡C stretch) was also detected in the decolorized melanin. The cytotoxicity study has shown that the treated melanin and LiP have low cytotoxic effects; however, the mediator of veratryl alcohol could result in high mortality which suggests that its use should be meticulously tested in formulating health and skincare products. The findings of the study suggest that LiP produced by Phanerochaete chrysosporium has the potential to be used in the medical and cosmetic industries, particularly for the development of biobased cosmetic whitening agents.

The purpose of this study was to synthesize a new magnetic material with antimicrobial properties, incorporated into a biopolymer and containing silver nanoparticles (Ag NP) prepared extract of Eugenia umbelliflora as a reducing agent. Silver nanoparticles incorporated into magnetic nanocomposite O-carboxymethylchitosan/y-Fe 2 O 3 /Ag 0 (CMAgE) composite were synthesized using an extract of E. umbelliflora. The antimicrobial activity of the pathogenic microorganism is reported here. The synthesized nanoparticles were also characterized, and quantified by Ag analysis. The minimum inhibitory concentrations (MIC) of CMAgE against Staphylococcus aureus, Escherichia coli, and Candida albicans were 16.5, 1000 and 500 μg/mL, respectively. The results show that these materials have significant synergistic effect on each other. The potential phytotoxic effect of the nanocomposites was evaluated using Cucumis sativus seeds. The positive values for seedling elongation inhibition (SEI) show that CMAgE and methanol extract of Eugenia umbelliflora (Eug) cause growth inhibition at a concentration of 1000 mg/L. The germination index (GI) values of 40% and 80% at 1000 mg/L, for CMAgE and Eug, respectively, showed inhibition of germination. CMAgE and Eug showed cytotoxic effects against Artemia salina nauplii, with LC 50 values of 72.5 μL/mL and < 5.0 μL/mL respectively, after 48 h., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The southern surf crab Ovalipes trimaculatus (de Haan, 1833) presents a high potential for aquaculture. In this study, we analyze the benefits of different dietary treatments on its molt success and fitness of larval stages. Artemia persimilis nauplii were enriched with monospecific (Nannochloropsis oculata, Tetraselmis suecica, Dunaliella salina, Isochrysis galbana and Chaetoceros gracilis) and multispecific (Mix) microalgal diets twice a day over a 48-h period. Mean total length (TL), growth instar number (I) and gut fullness rate (GFR) of nauplii showed significant differences between dietary treatments at several sampling times, optimal results being observed in those providing Mix. Artemia nauplii grown under most experimental dietary treatments reached the capture size limit for Ovalipes trimaculatus zoea I (700 µm) within 24 h. After that time interval, Mix-enriched nauplii were amongst those with higher protein contents. Ovalipes trimaculatus zoea I fed on Artemia nauplii enriched during 24 h under different dietary treatments showed significant differences in survival, inter-molt duration, molting success to zoea II and motility. Optimal results were observed in zoea I fed on Mix-enriched Artemia nauplii. This work not only represents a first step towards the dietary optimization for O. trimaculatus zoeae rearing but also provides the first results on the use of enriched A. persimilis.

The Metal Organic Framework (MOF) of formula {[Ag 6 (μ 3 -HMNA) 4 (μ 3 -MNA) 2 ] 2- ·[(Et 3 NH) + ] 2 ·(DMSO) 2 ·(H 2 O)} (AGMNA), a known efficient antimicrobial compound which contains the anti-metabolite, 2-thio-nicotinic acid (H 2 MNA), was incorporated in polymer hydrogels using, hydroxyethyl-methacrylate (HEMA). The material pHEMA@AGMNA-1 was characterized by X-ray fluorescence (XRF) spectroscopy, X-ray powder diffraction analysis (XRPD), Scanning Electron Microscopy (SEM), Energy-dispersive X-ray spectroscopy (EDX), Thermogravimetric Differential Thermal Analysis (TG-DTA), Differential Scanning Calorimetry (DTG/DSC), attenuated total reflection spectroscopy (FT-IR-ATR) and Ultrasonic Imaging. The antimicrobial capacity of pHEMA@AGMNA-1 was evaluated against the Gram negative bacterial strain Pseudomonas aeruginosa and the Gram positive ones of the genus of Staphylococcus epidermidis and Staphylococcus aureus, which are the etiology of the microbial keratitis. The % bacterial viability of P. aeruginosa, S. epidermidis and S. aureus upon their incubation with pHEMA@AGMNA-1 discs is significantly low (0.4 ± 0.1%, 1.5 ± 0.4% and 7.7 ± 0.5% respectively). The inhibition zones (IZ) caused by pHEMA@AGMNA-1 discs against P. aeruginosa, S. epidermidis and S. aureus are 14.0 ± 1.1, 11.3 ± 1.3 and 11.8 ± 1.8 mm respectively. Furthermore, pHEMA@AGMNA-1 exhibits low toxicity. Thus, pHEMA@AGMNA-1 might be an efficient candidate for the development of antimicrobial active contact lenses., Competing Interests: Declaration of competing interest The authors declare no any conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The Integrator complex (INT) contains several subunits that participate in RNAPII transcription and the 3' end process of non-coding RNAs. INTS11 is the catalytic subunit that interacts with the C-terminal domain of RNAPII, recently found to play a role in embryo development in different experimental models. However, the involvement of INTS11 is still ignorant in crustaceans, particularly in post-diapause embryonic development of Artemia sinica. In the present research, the full-length cDNA of As-Ints11 gene (1964 bp) was cloned from A. sinica by the RACE technique. The deduced 597 amino acids sequence contains the most identifiable domains of the INTs and is highly conserved. Immunofluorescence assay showed that the INTS11 was present at diverse developmental status in A. sinica: the As-INTS11 can be found in both cytoplasm and nucleus of the embryos, and the location showed no specificity in tissue or organ of the nauplius. The expression patterns of As-Ints11 were analyzed by qPCR and Western blotting, which show similar trends that peaked at the 15 h stage of embryo development. Moreover, the expressions of interacting proteins As-INTS9 and As-RNAPII were also detected, results display a synergetic effect with the As-INTS11 at both mRNA and protein levels. We also explored the amount of As-INTS11, As-INTS9 and As-RNAPII under different stresses, and the results indicate that the As-INTS11 is a stress-related protein though the mechanism needs further research. Knocking down of the As-INTS11 resulted in a delay of post-diapaused embryonic development in A. sinica., 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 © 2020 Elsevier B.V. All rights reserved.)

In the search for new drugs, strategies such as bioisosterism have been evidenced, in which the modification of molecules is already known to be active. Thus, metal complexes of known drugs have been highlighted, with examples of significant improvements in therapeutic efficacy. In this way, this work aimed at the synthesis of new zinc complexes with nonsteroidal anti-inflammatory drugs (NSAIDs), as well as the chemical characterization and the previous toxicity by cytotoxicity with Artemia salina, and evaluating the ability of these compounds to interact with DNA. As result, two new zinc II ternary complexes containing the NSAIDs diclofenac (Diclof) and ibuprofen (Ibup) and nicotinamide neutral linker (Nic) were obtained by the two-step solvent metal-ligand complexation method. Molecular structures were determined by NMR, FTIR, HR-MS, UV-Vis and X-ray diffraction, which demonstrated that both complexes are binuclear systems of general formula [Zn 2 (R-COO - ) 4 (Nic) 2 ]. Plasmidial DNA breakdown capacities were evaluated by producing single and double breaks (DNA FII and FIII) from plasmid incubation with complex solutions in the concentration range 0 to 400 μmol·L -1 in experiments with the presence and absence of light. Both experiments did not show significant differences (P ≤ 0.05) in induced DNA cleavage activity between the maximum study concentrations (400 μmol·L -1 ) and the negative controls for both complexes. The types of complex 1 and 2 interactions with the secondary DNA structure were determined by titrating a CT-DNA solution with complex solutions and monitored by circular dichroism spectrometry. The results showed that both complexes interact with the grooves of the secondary structure of CT-DNA by electrostatic attraction, but without evidence of alteration in the primary structure. Acute toxicity tests against Artemia salina showed that both complexes did not produce lethality >10% of the population up to a maximum concentration of 1200 μg·mL -1 within an exposure interval of 24 h. Thus, two new compounds were synthesized, characterized and had their previous toxicities determined. These compounds are promising new drugs, with the next step being evaluations of their activity., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Tributylin chloride (TBTCl) has been demonstrated to be acutely toxic to aquatic organisms., Aim: This study was conducted to investigate the effects of TBTCl on epithelial cell of gut Artemia salina in different stages (Nauplii, Juveniles, and Adults)., Methods: Samples of A. salina used were cultivated in incubators for hatching. Nauplii were harvested at 24 hours of age, while the juveniles and adults were harvested at 21 and 35 days of age, respectively. These three stages of A. salina were exposed to different concentrations of TBTCl (1 ng.L -1 to 500 ng.L -1 ) for 24 hours. For nauplii, juveniles, and adults, 100 individuals were exposed, and those that survived in the exposure test were harvested for histological analysis., Results: The histological examinations revealed significant differences ( p < 0.05) in type of lesions associated with different TBTCl concentrations and at different stages. The predominant lesions associated with different stages and different concentrations of TBTCl were epithelial cell necroasis, degeneration, cell loss, disruption, piknosis, and submucosal necrosis. Cell scoring was a significant difference ( p < 0.05) between the groups of different TBTCl concentrations and different life stages., Conclusion: Overall, in this study, the generality of the lesion scores showed that the adults are relatively more susceptible to the effects of TBTCl compared to the juvenile and the nauplii., Competing Interests: None

A mono-N-substituted probe L containing a bromosalicylaldehyde pendant arm attached to a tetraazamacrocyclic "tet a" moiety was synthesized via straight forward reaction. The probe L crystallizes in a monoclinic P2 1 /n space group. The probe L displayed quick sensitivity and selectivity towards Hg 2+ ions due to its hopeful Chelation Enhancement Quenching (CHEQ) feature. Interestingly, the probe L exhibits turn-off fluorescence response to Hg 2+ ion and turn-on fluorescence signals to HSO 4 - ions. When the probe L was complexed with HSO 4 - in 1:1 mode (L + HSO 4 - formation), improved turn-on fluorescence emission was detected due to the chelation enhanced fluorescence effect through sensor complex. The macrocyclic "tet a" probe L exhibited a binding constant value of 3.89 × 10 6  M -1 and 5.58 × 10 5  M -1 for Hg 2+ and HSO 4 - , respectively. Probe L exhibited good selectivity to Hg 2+ rather than other common metal ions and HSO 4 - over other common anions. The limit of detection (LOD) of Hg 2+ and HSO 4 - were found to be 1 nM and 7 μM, respectively. The time-resolved fluorescence emission single-photon counting study was used to determine the average lifetime value for the probe L and L + HSO 4 - ions as 0.47 and 1.02 ns, respectively. The practical application of the probe in visualizing intracellular Hg 2+ and HSO 4 - ions distribution in live Artemia salina was demonstrated. Furthermore, the probe L with Hg 2+ cations was found to be cytotoxic against breast cancer cells in nature and can be delivered as an anticancer agent. Besides the probe L with HSO 4 - exhibit strong fluorescence emission with low cytotoxicity, and it can be recommended for live-cell imaging., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: The cell cycle checkpoint protein RAD9 is a vital cell cycle regulator in eukaryotic cells. RAD9 is involved in diverse cellular functions by oligomer or monomer. However, the specific mechanism of its activity remains unknown in crustaceans, especially in embryonic diapause resumption of the brine shrimp Artemia sinica ., Methods and Results: In the present article, a 1238 bp full-length cDNA of As -RAD9 gene, encoding 376 amino acids, was obtained from A. sinica . The expression pattern of As -RAD9 was analyzed by qPCR and Western blot. The mRNA expression level climbs to the top at the 10 h stage of embryo development, while the protein expression pattern is generally consistent with qPCR results. Moreover, the As -RADd9 related signaling proteins, As -RAD1, As -HUS1, As -RAD17, and As -CHK1, were also detected. Immunofluorescence assay showed that the location of As -RAD9 did not show tissue or organ specificity, and the intracellular expression was concentrated in the cytoplasm more than in the nucleus. We also explored the amount of As -RAD9 under the stresses of cold and high salinity, and the results indicate that As -RAD9 is a stress-related factor, though the mechanisms may be different in response to different stresses. Knocking down of the As -RAD9 gene led to embryonic development delay in A. sinica ., Conclusions: All these results reveal that As -RAD9 is necessary for post-diapaused embryonic development in A. sinica .

Substituted phenyl[(5-benzyl-1,3,4-oxadiazol-2-yl)sulfanyl]acetates/acetamides 9a-j were synthesized as alkaline phosphatase inhibitors. Phenyl acetic acid 1 through a series of reactions was converted into 5-benzyl-1,3,4-oxadiazole-2-thione 4. The intermediate oxadiazole 4 was then reacted with chloroacetyl derivatives of phenols 6a-f and anilines derivatives 8a-d to afford the title oxadiazole derivatives 9a-j. All of the title compounds 9a-j were evaluated for their inhibitory activity against human alkaline phosphatise (ALP). It was found that compounds 9a-j exhibited good to excellent alkaline phosphatase inhibitory activity especially 9h displayed potent activity with IC 50 value 0.420 ± 0.012 µM while IC 50 value of standard (KH 2 PO 4 ) was 2.80 µM. The enzyme inhibitory kinetics of most potent inhibitor 9h was determined by Line-weaever Burk plots showing non-competitive mode of binding with enzyme. Molecular docking studies were performed against alkaline phosphatase enzyme (1EW2) to check the binding affinity of the synthesized compounds 9a-j against target protein. The compound 9h exhibited excellent binding affinity having binding energy value (-7.90 kcal/mol) compared to other derivatives. The brine shrimp viability assay results proved that derivative 9h was non-toxic at concentration used for enzyme assay. The lead compound 9h showed LD 50 106.71 µM while the standard potassium dichromate showed LD 50 0.891 µM. The DNA binding interactions of the synthesized compound 9h was also determined experimentally by spectrophotometric and electrochemical methods. The compound 9h was found to bind with grooves of DNA as depicted by both UV-Vis spectroscopy and cyclic voltammetry with binding constant values 7.83 × 10 3 and 7.95 × 10 3  M -1 respectively revealing significant strength of 9h-DNA complex. As dry lab and wet lab results concise each other it was concluded that synthesized compounds, especially compound 9h may serve as lead compound to design most potent inhibitors of human ALP., (Copyright © 2019 Elsevier Inc. All rights reserved.)

β-glucans, heterogeneous glucose polymers present in many organisms, have the capability to activate the innate immune system. Efficacy of activation depends on purity of the compound, molecular structure, polymerization degree, and source. One of the purest forms of crystallized β-(1-3)-glucan present in nature is the paramylon extracted from the WZSL non-chloroplastic mutant of Euglena gracilis , which can be processed to produce linear nanofibers capable of interacting with specific receptors present on cell membranes. The effects of these nanofibers, already investigated on plants, animals, and humans, will be analyzed., Competing Interests: The authors declare no conflicts of interest.

Although a substantial amount of research exists on pollution and biological invasions, there is a paucity of understanding of how both factors interact. Most studies show that pollution favours the establishment of invasive species, but pollution may also promote local adaptation of native species and prevent the establishment of new incomers. However, evidence for this is extremely limited because most studies focus on successful invasions and very few on cases where an invasion has been resisted. Here we provide evidence of local adaptation of native species to pollution combining life history and physiological data. We focused on the invasion of the North American brine shrimp Artemia franciscana, which is causing a dramatic biodiversity loss in hypersaline ecosystems worldwide, and one of the last native Artemia populations in SW Europe (A. parthenogenetica from the historically polluted Odiel estuary, SW Spain). Life table response experiments were carried out in the laboratory to compare the demographic responses of A. parthenogenetica and a nearby A. franciscana population to long-term Zn exposure (0.2 mg L -1 ). We also evaluated oxidative stress by measuring antioxidant defences (catalase, glutathione reductase and superoxide dismutase) and lipid peroxidation (thiobarbituric acid reactive substances). A high concentration of Zn induced strong mortality in A. franciscana, which also showed high levels of lipid peroxidation, suggesting relatively poor physiological resistance to pollution compared with A. parthenogenetica. The age at maturity was shorter in A. parthenogenetica, which may be an adaptation to the naturally high mortality rate observed in the Odiel population. Exposure to Zn accelerated age at first reproduction in A. franciscana but not in A. parthenogenetica. In contrast, Zn had a stimulatory effect on offspring production in A. parthenogenetica,which also showed higher reproductive parameters (number of broods, total offspring and offspring per brood) than A. franciscana. Overall, the results of this study strongly suggest that native Artemia from Odiel estuary is locally adapted (at both, reproductive and physiological levels) to Zn contamination and that A. franciscana is highly sensitive. This is a good example of how pollution may play a role in the persistence of the last native Artemia populations in the Mediterranean., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The ability of some microorganisms to use clarified cashew apple juice as carbon and energy source for biosurfactant production was assessed under strict controlled conditions. Twelve strains of Bacillus were isolated and evaluated regarding their biosurfactant production capabilities. The biosurfactant obtained with these selected strains showed the capacity of decreasing the surface tension of water from 72.0 to 31.8 mN.m -1 and the interfacial tension of n-hexadecane to 27.2 mN.m -1 , with a critical micelle concentration of 12.5 mg.L -1 . Not only did the biosurfactant present excellent stability to pH, temperature and salinity, it also showed emulsifying properties in different hydrocarbons. The behavior of the phase diagrams showed the potential of the produced biosurfactant to obtain relatively-stable emulsions for up to 96 h, which allows for its application in several areas. The semi-purified biosurfactant did not show toxicity against Lactuca sativa (lettuce) or Artemia salina (microcrustacean), presenting an LC 50 of 612.27 μ mL -1 . The surfactant was characterized as being a cyclic lipopeptide with molecular structure similar to that of surfactin. Furthermore, through the employment of the surfactant produced, the remediation effect in oil-contaminated soil could be significantly improved., (Copyright © 2018. Published by Elsevier B.V.)

This study deals with synthesis of chromium nanoparticles (CrNPs) from potassium dichromate using the aqueous extract of Allium sativum. They were characterized through UV-VIS light, FE-SEM, EDX, XRD, and FT-IR, which revealed uniform, mono-dispersive, and highly stable CrNPs of 31-64-nm size. The Artemia nauplii was enriched with 4.94 mg/L of CrNPs (24-h LC 50 ) at different durations (½, 1, 2, and 4 h) and then fed to Macrobrachium rosenbegii post-larvae (PL) for 30 days as live feed. The results showed that ½- and 1-h enriched Artemia nauplii led to significant improvements in nutritional indices including growth and survival, and concentrations of tissue biochemical constituents, such as total protein, amino acid, carbohydrate, and lipid of M. rosenbergii PL (P < 0.05), which suggests that this concentration of CrNPs was non-toxic to M. rosenbergii PL. This was confirmed by the insignificant alterations recorded in activities of SOD and CAT (P > 0.05) in M. rosenbergii PL fed with ½- and 1-h enriched Artemia nauplii as live feed. After that, SOD and CAT activities started to increase. Therefore, this optimized concentration of CrNPs (4.94 mg/L) is recommended for enrichment of Artemia nauplii for ½-1-h duration as a sustainable material in the nursery of M. rosenbergii.

Eighty cultures from the Novi Sad Cyanobacterial Culture Collection (NSCCC) were screened for toxicity with Artemia salina bioassay and for common cyanobacterial toxins, microcystins/nodularin (MCs/NOD) and saxitoxin (STX), with ELISA assays. The results show that 22.5% (11) of the investigated cyanobacterial cultures in exponential phase exhibited toxicity in the A. salina bioassay and 38.7% (31) produced MCs/NOD and/or STX. However, the findings in the two methods applied were contradictory. Therefore, A. salina bioassay was repeated on 28 cultures in stationary growth phase, which were positive in ELISA assays but not in the initial A. salina bioassay. Seven more cultures exhibited cell-bound toxicity, and only one extracellular toxicity. The observed difference in the toxicity indicates that cyanobacterial growth phase could affect the screening results. The findings also varied depending on the environment from which the cultures originated. In the initial screening via bioassay, 11.8% (6 cultures out of 51) from terrestrial and 17.2% (5 out of 29) from aquatic environment showed cell-bound toxicity. Furthermore, based on the ELISA assay, 31.4% (16) of the cultures from terrestrial ecosystems were positive for the presence of the investigated cyanotoxins, and 51.7% (15) from aquatic ecosystems. Based on all results, more frequent toxin production was observed in cultures originating from aquatic environments. Furthermore, the group of terrestrial cultures that originated from biological loess crusts were basically non-toxic. The discrepancies in the results by two different methods indicates that the use of several complementary methods would help to improve the assessment of cyanobacterial toxicity and cyanotoxin analyses., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This study investigated four different environmentally relevant microplastic (MP) pollutants which were derived from two facial cleansers, a plastic bag and polyethylene textile fleece. The mean size range of the particles (according to number distribution) was 20-250 μm when measured as a powder and 0.02-200 μm in suspension. In all MP exposures, plastic particles were found inside the guts of D. magna and A. franciscana, but only in the case of daphnids a clear exponential correlation between MP uptake in the gut and the size of the MP was identified. Exposure tests in which the majority of the MP particles were below 100 μm in size also had higher numbers of daphnids displaying evidence of MP ingestion. As the average MP particle size increased, the percentage of daphnids which had MP in their gut decreased. Using a number distribution value to measure particle size when in a suspension is more experimentally relevant as it provides a more realistic particle size than when samples are measured as a powder. Generally, artemias had fewer MP particles in the gut, than the daphnids, which could be explained by their different food size preferences. No acute effects on D. magna were found, but the growth of A. franciscana was affected. We conclude that zooplankton crustacean can ingest various MPs but none of the exposures tested were highly acutely hazardous to the test species. In addition, no delayed lethal effects in a 24 h post-exposure period were found., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The present study was aimed to investigate the antibacterial efficacy of fucoidan mediated silver nanoparticles (Fu-AgNPs) synthesized from Spatoglossum asperum. The synthesized Fu-AgNPs were characterized by UV-visible, Field emission - scanning electron microscope (FE-SEM), Tranmission electron microscope (TEM), X-ray diffraction (XRD), Selected area electron diffraction (SAED) pattern, Energy-dispersive X-ray spectroscopy (EDAX), Fourier transform infrared spectroscopy (FT-IR), Dynamic light scattering (DLS) and Zeta potential analysis. The UV-visible spectrum of Fu-AgNPs exhibited a characteristic surface plasmon resonance (SPR) peak at 440 nm. The electron microscopic results revealed that the nanoparticles were spherical to oval in shape and are found to be 20 to 46 nm. Altogether the X-ray diffraction analysis showed that the Fu-AgNPs were crystalline in nature. The FT-IR spectrum confirmed the existence of CC stretching vibration of aromatic compounds and sulfated groups of fucoidan plays a major role in the synthesis of Fu-AgNPs. The biosynthesized Fu-AgNPs shows potential antibacterial activity against Klebsiella pneumoniae in agar bioassay, disk diffusion, reactive oxygen species, protein leakage and confocal laser scanning microscopy assays. Furthermore, Artemia toxicity assay results showed less mortality (3.3 ± 0.8%) even at higher concentration of Fu-AgNPs. Therefore, Fu-AgNPs can be effectively used as an antibacterial agent in the pharmaceutical fields., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this study, we evaluated a consortium of probiotic bacteria as an environmentally-friendly strategy for controlling pathogenic Vibrio species during the brine shrimp incubation period. Probiotic strains were initially selected on basis of (i) their ability to colonize the cyst surfaces, (ii) their absence of cross-inhibitory effects, and (iii) no detrimental effect on cyst hatching. The cysts and nauplius surfaces were immediately colonized after the application of selected probiotic strains, without detrimental effects on survival. Ten probiotic strains were mixed at similar proportions (probiotic consortium) and evaluated at different concentrations into brine shrimp cultures during incubation and early stages of development. Subsequently, these cultures were challenged with Vibrio parahaemolyticus and Vibrio harveyi. The probiotic consortium was effective to reduce the abundance of pathogenic Vibrio species and to prevent the mortality during Vibrio challenges; however, its effect was concentration-dependent and was successful at a starting concentration of 1.8 × 10 6  CFU/ml. Our results suggest that this probiotic consortium offers an alternative to antimicrobial agents routinely used to reduce the incidence and prevalence of pathogenic Vibrio species in brine shrimp production.

Post-diapause cysts of Artemia franciscana undergo a well-defined developmental process whereby internal differentiation leads to rupture of the cyst shell, release of membrane-enclosed nauplii and hatching to yield swimming larvae. The post-diapause development of A. franciscana has been examined at biochemical and molecular levels, yet little is known about molecular chaperone function during this process. In addressing this we recently described ArHsp40, a type 1 J-domain protein in post-diapause A. franciscana cysts and larvae. The current report describes ArHsp40-2, a second J-domain protein from A. franciscana. ArHsp40-2 is a type 2 J-domain protein, lacking a zinc binding domain but containing other domains characteristic of these proteins. Notably, ArHsp40-2 possesses a double barrel β-domain structure in its substrate binding region, as does ArHsp40. qPCR revealed a relatively low amount of ArHsp40-2 mRNA in 0 h cysts which increased significantly until the E1 stage, most likely as a result of enhanced transcription, after which it declined. An antibody specific to ArHsp40-2 was produced and used to show that like its mRNA, ArHsp40-2 accumulated until the E1 stage and then decreased to amounts lower than those in 0 h cysts. The synthesis of ArHsp40-2 was induced by heat shock indicating that ArHsp40-2 is involved in stress resistance in cysts and nauplii. Accumulation in cysts during early post-diapause development followed by its sharp decline suggests a role in protein disaggregation/refolding, a function of Hsp40s from other organisms, where ArHsp40-2 assists in the rescue of proteins sequestered during diapause by p26, an abundant small heat shock protein (sHsp) in A. franciscana cysts., Competing Interests: The authors have declared that no competing interests exist.

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%.

The brine shrimp Artemia was used as a model organism to test toxicity of several neuroactive pesticides (chlorpyrifos (CLP), chlorpyrifos oxon (CLP ox), diazinon (DZN), carbaryl (CBR)) following exposure to far below than lethal doses. Cysts were exposed to the pesticides in order to test a scenario similar to actual coastal environment contamination, by analyzing different responses. Cysts were rehydrated in water containing the pesticides at concentrations ranging from 10 -11 to 10 -5  M, for 72, 96 and 192 h, respectively. For these exposure times, morpho-functional and biochemical parameters, such as hatching speed and viability were investigated in the larvae together with cholinesterase (ChE) activity quantification and histochemical localization. Finally, ChE inhibition was also compared with conventional selective ChE inhibitors. Results showed that CLP ox and CBR caused a significant dose-dependent decrease in hatching speed, followed by high percentages of larval death, while CLP and DZN were responsible for irregular hatching patterns. In addition, the pesticides mostly caused larval death some days post-hatching, whereas this effect was negligible for the specific ChE inhibitors, suggesting that part of pesticide toxicity may be due to molecules other than the primary target. ChE activity was observed in the protocerebrum lobes, linked to the development of pair eyes. Such activity was inhibited in larvae exposed to all pesticides. When compared to conventional selective inhibitors of ChE activities, this inhibition demonstrated that the selected pesticides mainly affect acetylcholinesterase and, to a lesser extent, pseudocholinesterases. In conclusion, the brine shrimp is a good model to test the environmental toxicity of long term exposure to cholinergic pesticides, since changes in hatching speed, viability and ChE activity were observed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A series of quinoline based peptides were synthesized by a one-pot reaction through Ugi-four component condensation of lipoic acid, cyclohexyl isocyanide, aniline derivatives and 2-methoxy quinoline-3-carbaldehyde derivatives under microwave irradiation. The products were obtained in excellent yields and high purity. Solvent optimization and the effect of microwave irradiation with various powers were also observed. All the synthesized compounds were characterized by FTIR, NMR spectral data and elemental analysis. A total of eight peptides were subjected to antimicrobial, antioxidant and toxicity evaluation. Among them, four peptides showed potential towards antibacterial screening with Bacillus cereus, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis and Candida albicans, Candida utilis and three peptides showed antioxidant test positive (DPPH). Besides, toxicity of all the peptides were evaluated by using brine shrimp and it was observed that four peptides showed mortality rate less than 50% up to 48h. Molecular docking studies revealed that the higher binding affinity of the two peptides toward DNA gyrase than ciprofloxacin based on Libdock score. The described chemistry represents a facile tool to synthesize complex heterocycles of pharmaceutical relevance in a highly efficient and one-pot fashion. The advantages of this method are its green approach, inexpensive solvent, shorter reaction times and excellent yields., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The square planar Pt(II) complexes of the type [Pt(L n )(Cl 2 )] (where L n  = L 1-3  = thiophene-2-carboxamide derivatives and L 4-6  = thiophene-2-carbothioamide derivatives) have been synthesized and characterized by physicochemical and various spectroscopic studies. MIC method was employed to inference the antibacterial potency of complexes in reference to free ligands and metal salt. Characteristic binding constant (K b ) and binding mode of complexes with calf thymus DNA (CT-DNA) were determined using absorption titration (0.76-1.61 × 10 5  M -1 ), hydrodynamic chain length assay and fluorescence quenching analysis, deducing the partial intercalative mode of binding. Molecular docking calculation displayed free energy of binding in the range of -260.06 to -219.63 kJmol -1 . The nuclease profile of complexes towards pUC19 DNA shows that the complexes cleave DNA more efficiently compared to their respective metal salt. Cytotoxicity profile of the complexes on the brine shrimp shows that all the complex exhibit noteworthy cytotoxic activity with LC 50 values ranging from 7.87 to 15.94 μg/mL. The complexes have been evaluated for cell proliferation potential in human colon carcinoma cells (HCT 116) and IC 50 value of complexes by MTT assay (IC 50  = 125-1000 μg/mL).

Artemia diapause has been extensively studied in embryonic biology for a long time. It has been demonstrated that hydrogen peroxide (H 2 O 2 ) can increase the hatching rate in the development and diapause termination of Artemia cysts. This study used an untargeted 1 H NMR-based metabolomic approach to explore the physiological regulation of H 2 O 2 in initiating the development and terminating the diapause of Artemia cysts. This experiment was divided into two parts. In the first part, we analyzed three groups with or without H 2 O 2 as control-0h, control-5h and H 2 O 2 (180μM)-5h; in the second part, after 7-d incubation, the non-hatching cysts were treated with different H 2 O 2 concentrations as low as 180μM and as high 1800μM. The results showed that arginine and proline metabolism were up-regulated after 5h, and H 2 O 2 up-regulated valine, leucine and isoleucine biosynthesis in the development of cysts. In the second part, low H 2 O 2 (180μM) showed alanine, aspartate and glutamate metabolism, but high H 2 O 2 (1800μM) also up-regulated arginine and proline metabolism, as in the control group without H 2 O 2 stimulus. These results suggest that enough H 2 O 2 can catalyze cell transcription and translation in Artemia cysts, and it improves the cell growth rate, thus allowing embryo cells to grow again., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Coolia are marine benthic dinoflagellates which are globally distributed and potentially toxic. This study provides the first investigation of species diversity and toxicity assessment of Coolia in Hong Kong waters. Fifty-one strains of four Coolia species, including C. malayensis, C. canariensis, C. tropicalis, and C. palmyrensis, were isolated from twelve sub-tidal habitats, and identified phylogenetically using 28S rDNA sequences. Exposure experiments (48-hour) demonstrated that the algal lysates extracted from the four Coolia species exhibited different toxic effects on the lethality and abnormality of two invertebrate larvae, i.e., brine shrimp Artemia franciscana and sea urchin Heliocidaris crassispina. Heliocidaris crassispina was more sensitive to the toxic effects of Coolia species than A. franciscana. Toxicity tests from both larvae revealed that C. malayensis was generally more toxic, and caused higher mortality rates when compared with the other three species. The emerging threat of harmful benthic dinoflagellates to marine environments and sensitive biota is discussed., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Using Artemia salina cysts (capsulated and decapsulated) and larvae (instar I, II and III) as experimental models, the potential effects of Fe 3 O 4 nanoparticles (Fe 3 O 4 -NPs) on marine ecosystems were investigated. Hatchability, mortality and a number of ethological, morphological and biochemical parameters were selected as end-points to define the toxic responses. Data showed that the hatching rates of capsulated and decapsulated cysts were significantly decreased (p < 0.01) following exposure to 600 mg/L for 24 and 36 h. The LC 50 values for instar II and III were 482 and 561 mg/L (could not be measured for instar I), and the EC 50 values for swimming inhibition of instar I, II and III were 474, 365 and 421 mg/L, respectively. Effects on hatchability, mortality and swimming were accounted for Fe 3 O 4 -NPs rather than iron ion released from the NPs. Instar II larvae showed the greatest sensitivity to Fe 3 O 4 -NPs, and followed by instar III, instar I, decapsulated cysts and capsulated cysts. Body lengths of instar I, II and III larvae were decreased in dose-dependent manners. Fe 3 O 4 -NPs attached onto the gills 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 substantially increased following exposure, indicating that toxic effects were related to oxidative stress. Mitochondrial malformation, cristae rupturing and membranous structure disruption were clearly observed after Fe 3 O 4 -NPs exposure. Fe 3 O 4 -NPs were ingested and well distributed in the gut, yolk and primary body cavity. Uptake kinetics data showed that the maximum Fe 3 O 4 -NPs content (16.4 mg/g) was reached at 30 h. The combined results so far indicate that Fe 3 O 4 -NPs have the potential to affect aquatic organisms when released into the marine ecosystems., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Controlling disease outbreaks is a major challenge in aquaculture farms and conventional methods are often ineffective. Nutritional supplementation and probiotic preparations help in reducing severity of such infections. The generally regarded as safe yeast (Yarrowia lipolytica) was used in the current study. A marine strain of Y. lipolytica exhibited tolerance towards sodium selenite and formed cell associated selenium nanoparticles (SeNPs). The synthesized nanoparticles were characterized by UV-vis spectroscopy, X-ray diffraction (XRD) and Field Emission Scanning Electron Microscope (FE-SEM) observations. Fourier transform infra-red (FTIR) spectroscopy indicated the role of carboxylic and amine groups in the synthesis of nanoparticles. This SeNP-enriched biomass was used as feed for the model aquaculture system, Artemia salina and compared with normal feed, baker's yeast (Saccharomyces cerevisiae). A. salina fed with SeNP-enriched biomass, showed increased survival rates (96.66%) as compared to those fed with S. cerevisiae (60.0%). The size of the larvae fed with SeNP-enriched biomass of Y. lipolytica was also found to be larger. Additionally, larval groups fed with SeNP-enriched biomass were better protected (70.0% survival) against V. harveyi infection when compared with groups fed with S. cerevisiae (24.44%). This combination of selenium in the nanoparticle form associated with the biomass of Y. lipolytica has potential application in improving health of aquaculture species in farms., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: The consumption of wild mushrooms has increased in recent years. However, not all mushrooms are edible and some of them may cause poisoning. Therefore, their toxicity needs to be studied. Artemia franciscana is a crustacean used in toxicity tests including toxins of fungi., Aims: To determine the percentage of inhibition and mortality produced by extracts of several basidiomycetes on the hatching of A. franciscana cysts., Methods: Aqueous extracts were prepared from 15 species of mushrooms collected from Jalisco state, Mexico. Different concentrations of the extracts were assayed in order to test their toxicity. Potassium dichromate and artificial seawater were the positive and negative controls, respectively. The percentages of hatching and mortality of the cysts were evaluated., Results: Inhibition of hatching greater than 80% in all the concentrations tested was found in 13 of the 15 species studied, in contrast to the positive control, which inhibited cyst hatching less than 50% in all cases. The highest percentage of mortality in the cysts was caused by the aqueous extracts of Amanita virosa, Leucopaxillus amarus, and Tylopilus violatinctus, and the lowest by Macrolepiota mastoidea., Conclusions: The brine shrimp bioassay appeared to be useful in the evaluation of the toxicity of several basidiomycetes, with the exception of Scleroderma texense, a mushroom considered poisonous, which showed no toxicity over A. franciscana., (Copyright © 2017 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.)

The extensive use and release to the aquatic environment of silver nanoparticles (NPs) could lead to their incorporation into the food web. Brine shrimp larvae of 24h showed low sensitivity to the exposure to PVP/PEI-coated Ag NPs (5nm), with EC 50 values at 24h of 19.63mgAgL -1 , but they significantly accumulated silver after 24h of exposure to 100μgL -1 of Ag NPs. Thus, to assess bioaccumulation and effects of silver transferred by the diet in zebrafish, brine shrimp larvae were exposed to 100ngL -1 of Ag NPs as an environmentally relevant concentration or to 100μgL -1 as a potentially effective concentration and used to feed zebrafish for 21days. Autometallography revealed a dose- and time-dependent metal accumulation in the intestine and in the liver of zebrafish. Three-day feeding with brine shrimps exposed to 100ngL -1 of Ag NPs was enough to impair fish health as reflected by the significant reduction of lysosomal membrane stability and the presence of vacuolization and necrosis in the liver. However, dietary exposure to 100μgL -1 of Ag NPs for 3days did not significantly alter gene transcription levels, neither in the liver nor in the intestine. After 21days, biological processes such as lipid transport and localization, cellular response to chemical stimulus and response to xenobiotic stimulus were significantly altered in the liver. Overall, these results indicate an effective dietary transfer of silver and point out to liver as the main target organ for Ag NP toxicity in zebrafish after dietary exposure., (Copyright © 2017 Elsevier Inc. All rights reserved.)

In this study, the toxicity of methylparaben (MeP) an emerging contaminant, was analysed in the sexual species Artemia franciscana, due to its presence in coastal areas and marine saltworks in the Mediterranean region. The acute toxicity (24h-LC 50 ) of MeP in nauplii was tested and its chronic effect (9days) evaluated by measuring survival and growth under two sublethal concentrations (0.0085 and 0.017mg/L). Also, the effect on several key enzymes involved in: antioxidant defences (catalase (CAT) and gluthathion-S-transferase (GST)), neural activity (cholinesterase (ChE)) and xenobiotic biotransformation (carboxylesterase (CbE), was assessed after 48h under sublethal exposure. The results of acute exposure indicate that MeP is harmful to A. franciscana (24h-LC 50 =36.7mg/L). MeP causes a decrease in CAT activity after 48h exposure to both concentration tested, that points out at the oxidative stress effect of MeP in A. franciscana. However, no significant effect on ChE, CbE and GST activities was found. In addition, MeP does not affect survival and growth in chronic exposure at the sublethal concentrations tested. The results of this study indicate that MeP is not a threat for A. franciscana under the experimental conditions used. Additional studies should be done considering long-term exposure and reproduction studies to analyse the potential risk of MeP as emerging contaminant in marine and hypersaline environments., (Published by Elsevier Inc.)

Selenium is an essential microelement for the normal functioning of life processes. Moreover, it is a component of enzymes with antioxidant effects. However, it has the smallest window of any micronutrient between requirement and toxicity. Selenium is a regularly used element in fish feeds; moreover, enriching zooplankton with selenium to rear larvae is also a well-known technology. It is accepted that the most common starter foods of fish larvae, natural rotifers contain the smallest dosage of selenium, but providing selenium enriched Artemia sp. instead could increase survival and growth rate of fish. However, no such references are available for the red drum (Sciaenops ocellatus) larvae. Therefore, in this study, Artemia sp. was enriched with nano-selenium of verified low toxicity and easy availability in 5 treatments (1, 5, 10, 50, 100 mg/l Se), and then, fish larvae were fed with four of these enriched Artemia stocks (1, 5, 10, 50 mg/l Se) and a control group. At the end of the 9-day-long experiment, survival rate (S) and growth parameters (SL, W, K-factor, SGR) of fish larvae were calculated as well as their selenium retention and glutathione peroxidase enzyme activity were analysed. It was revealed that a moderate level of selenium enrichment (~4 mg/kg dry matter) of Artemia sp. positively influences the rearing efficiency (i.e. survival and growth) of fish larvae, but higher dosages of selenium could cause adverse effects.

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.

Glyphosate-based herbicides are the most commonly used worldwide because they are effective and relatively nontoxic to nontarget species. Unlimited and uncontrolled use of such pesticides can have serious consequences for human health and ecological balance. The present study evaluated the acute toxicity and genotoxicity of 2 glyphosate-based formulations, Roundup Original (Roundup) and Glyphosate AKB 480 (AKB), on different organisms: cucumber (Cucumis sativus), lettuce (Lactuca sativa), and tomato (Lycopersicon esculentum) seeds, and microcrustacean Artemia salina and zebrafish (Danio rerio) early life stages. For the germination endpoint, only L. esculentum presented significant sensitivity to AKB and L. sativa to Roundup, whereas both formulations significantly inhibited the root growth of all species tested. Both AKB and Roundup induced significant toxicity to A. salina; both are classified as category 3, which indicates a hazard for the aquatic environment, according to criteria of the Globally Harmonized Classification System. However, Roundup was more toxic than AKB, with 48-h median lethal concentration (LC50) values of 14.19 mg/L and 37.53 mg/L, respectively. For the embryo-larval toxicity test, Roundup proved more toxic than AKB for the mortality endpoint (96-h LC50 values of 10.17 mg/L and 27.13 mg/L, respectively), whereas for the hatching parameter, AKB was more toxic than Roundup. No significant genotoxicity to zebrafish larvae was found. We concluded that AKB and Roundup glyphosate-based formulations are phytotoxic and induce toxic effects in nontarget organisms such as A. salina and zebrafish early life stages. Environ Toxicol Chem 2017;36:1755-1763. © 2016 SETAC., (© 2016 SETAC.)

Phytol (PYT) is a diterpenoid having important biological activity. However, it is a water non-soluble compound. This study aims to prepare PYT nanoemulsion (PNE) and evaluation of toxic, cytotoxic and genotoxic activities of PYT and PNE. For this, the PNE was prepared by the phase inversion method. The cytotoxicity test was performed in Artemia salina, while toxicity, cytotoxicity and genotoxicity in Allium cepa at concentrations of 2, 4, 8 and 16 mM. Potassium dichromate and copper sulfate were used as positive controls for the tests of A. salina and A. cepa, respectively. In addition, an adaptation response was detected in A. cepa by using the comet assay. The results suggest that both PYT and PNE exhibited toxic and cytotoxic effects at 4-16 mM in either test system, while genotoxicity at 2-16 mM in A. cepa. PNE exhibited more toxic, cytotoxic and genotoxic effects at 8 and 16 mM than the PYT. However, both PYT and PNE at 2 and 4 mM decreased the index and frequency of damage in A. cepa after 48 and 72 h, suggesting a possible adaptation response or DNA damage preventing capacity. Nanoemulsified PYT (PNE) may readily cross the biological membranes with an increase in bioavailability and produce more toxic, cytotoxic and genotoxic effects in the used test systems., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Induction of HSP72 is a natural response of stressed organisms that protects against many insults including bacterial diseases in farm (aquatic) animals. It would therefore be of great health benefit to search for natural compounds that are clinically safe yet able to induce HSP72 in animals. The phenolic compound carvacrol, an approved food component, had been shown in in vitro study to act as a co-inducer of HSP72, enhancing HSP72 production only in combination with a bona fide stress compared to the compound alone. However, in vitro model systems do not completely represent an in vivo physiology. Here, using the well-established gnotobiotic Artemia model system, we determined whether carvacrol could induce HSP72 in vivo, whether this putative effect could generate resistance in Artemia against biotic/abiotic stress and also unraveled the mechanism behind the possible HSP72-inducing effect of carvacrol. The gnotobiotic system is crucial for such studies because it avoids the interference of any extraneous factors on host-compound interaction. Here, carvacrol was shown to be a potent HSP72 inducer. Induction of HSP72 was associated with the generation of resistance in Artemia larvae against subsequent lethal heat stress or pathogenic Vibrio harveyi. Our results also provided new insight on the mode of HSP72 inducing action of carvacrol, in which the initial generation of reactive molecule H 2 O 2 by the compound plays a key role. Overall results add new information about the bioactivity of carvacrol and advance our knowledge of this compound as potential prophylactic agent for controlling Vibrio infection in aquaculture animals.

Zinc oxide (ZnO) nanostructures are synthesized in various organic solvents (acetone, chloroform, ethyl acetate, ethanol and methanol) and water via coprecipitation process using zinc acetate as precursor. The resultant ZnO nanoparticles, nano rods and nano sheets are characterized by UV-vis spectrophotometric analysis, scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transmission infrared spectroscopy (FTIR), and energy dispersive X-ray spectroscopy (EDX). The variable size and geometry of nanoparticles depend upon medium used for synthesis. The synthesized ZnO nanostructures exhibit minor to moderate antioxidative (DPPH based free radical scavenging activity, total antioxidative potential and total reducing power) response. Mild to moderate antibacterial and antifungal activities, excellent antileishmanial potential (IC50 up to 3.76), and good cytotoxic perspective (LD50 up to 49.4) is also observed by the synthesized ZnO NPs. The nanoparticles also exhibit moderate α-amylase inhibition response. Furthermore the nanostructures are evaluated for methylene blue photodegradation response within 60min time period. It is found that organic solvent alters shape, size and other physio-chemical properties of ZnO that ultimately modulate the biological, chemical, and environmental properties., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Six new compounds including two furanone derivatives sclerotiorumins A and B (1 and 2), one novel oxadiazin derivative sclerotiorumin C (3), one pyrrole derivative 1-(4-benzyl-1H-pyrrol-3-yl)ethanone (4), and two complexes of neoaspergillic acid aluminiumneohydroxyaspergillin (5) and ferrineohydroxyaspergillin (6) were isolated from the co-culture of marine-derived fungi Aspergillus sclerotiorum and Penicillium citrinum. Compound 3 was the first natural 1,2,4-oxadiazin-6-one. Compound 5 showed significant and selective cytotoxicity against human histiocytic lymphoma U937 cell line (IC 50  = 4.2 μm) and strong toxicity towards brine shrimp (LC 50  = 6.1 μm), and oppositely increased the growth and biofilm formation of Staphylococcus aureus., (© 2017 Wiley-VHCA AG, Zurich, Switzerland.)

There have been many studies on dopamine active transporter (DAT) in humans and laboratory animals; however, there is a lack of information on DAT in brine shrimp. In this study, we demonstrated the neuronal and nonneuronal characteristics of DAT-synthesizing (DAT + cells) during development of brine shrimp. In neuronal cells, the DAT + neurons in the central body and lobes of a protocerebrum (PC) controlled the deutocerebrum. The sensory cells of nauplius eyes projected their decussated axons to the PC, and the DAT + cells at the posterior region were associated with migration and control of the 10 posterior neurons during the early nauplius stage. In nonneuronal cells, the five types of glands, that is, the salt, antennal, mandible, and accessory glands and posterior gland1 and gland2 synthesized DAT protein. In addition, the gut and rectum dilator muscles and renal cells expressed DAT protein. Thus, DAT protein acts in the development of several types of cells during development of brine shrimp., (© 2017 Wiley Periodicals, Inc.)

A novel method, BBD-SSPD is proposed by the combination of Box-Behnken Design (BBD) and Split-Split Plot Design (SSPD) which would ensure minimum number of experimental runs, leading to economical utilization in multi- factorial experiments. The brine shrimp Artemia was tested to study the combined effects of photoperiod, temperature and salinity, each with three levels, on the hatching percentage and hatching time of their cysts. The BBD was employed to select 13 treatment combinations out of the 27 possible combinations that were grouped in an SSPD arrangement. Multiple responses were optimized simultaneously using Derringer's desirability function. Photoperiod and temperature as well as temperature-salinity interaction were found to significantly affect the hatching percentage of Artemia, while the hatching time was significantly influenced by photoperiod and temperature, and their interaction. The optimum conditions were 23 h photoperiod, 29 °C temperature and 28 ppt salinity resulting in 96.8% hatching in 18.94 h. In order to verify the results obtained from BBD-SSPD experiment, the experiment was repeated preserving the same set up. Results of verification experiment were found to be similar to experiment originally conducted. It is expected that this method would be suitable to optimize the hatching process of animal eggs.

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.)

Upon diapause termination and exposure to favorable environmental conditions, cysts of the crustacean Artemia franciscana reinitiate development, a process dependent on the resumption of metabolic activity and the maintenance of protein homeostasis. The objective of the work described herein was to characterize molecular chaperones during post-diapause growth of A. franciscana. An Hsp40 complementary DNA (cDNA) termed ArHsp40 was cloned and shown to encode a protein with an amino-terminal J-domain containing a conserved histidine, proline, and aspartic acid (HPD) motif. Following the J-domain was a Gly/Phe (G/F) rich domain, a zinc-binding domain which contained a modified CXXCXGXG motif, and the carboxyl-terminal substrate binding region, all characteristics of type I Hsp40. Multiple alignment and protein modeling showed that ArHsp40 is comparable to Hsp40s from other eukaryotes and likely to be functionally similar. qRT-PCR revealed that during post-diapause development, ArHsp40 messenger RNA (mRNA) varied slightly until the E2/E3 stage and decreased significantly upon hatching. The immunoprobing of Western blots demonstrated that ArHsp40 was also relatively constant until E2/E3 and then declined dramatically. The drop in ArHsp40 when metabolism and protein synthesis were increasing was unexpected and demonstrated developmental regulation. The reduction in ArHsp40 at such an active life history stage indicates, as one possibility, that A. franciscana possesses additional Hsp40s, one or more of which replaces ArHsp40 as development progresses. Increased synthesis upon heat shock established that in addition to being developmentally regulated, ArHsp40 is stress inducible and, because it is found in mature cysts, ArHsp40 has the potential to contribute to stress tolerance during diapause.

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.)

Parthenogenetic Artemia from seven Chinese locations with different elevations and various ploidies are characterized by phenotypic and morphometric analyses. Our findings show that the studied populations exhibit dissimilar patterns of ovisac. Four phenotypic patterns of furca are qualified and one of them is shared among di-, tetra- and pentaploid Artemia. Results of discriminant analysis based on morphometric data reveal that tetra- and pentaploid populations are grouped together, but the Aqqikkol Lake population is clearly differentiated. Previous hypothesis/conclusion that polyploid Artemia are larger than diploids is only partly supported by the present results, which show that pentaploid and tetraploid populations are larger than the mostly diploid populations in terms of the total length, but the body size of the Aibi Lake triploids has not significant difference with the sympatric diploids and the mostly diploid Aqqikkol population that inhabit in very high altitude has the largest body size among all parthenogenetic populations. The founding confirms that body size of Artemia is following with Bergmann's rule., (© 2016 Wiley Periodicals, Inc.)