Transcriptomic analysis of Neocaridina denticulate sinensis hepatopancreas indicates immune changes after copper exposure.

Neocaridina denticulate sinensis is a promising crustacean model species due to its merits in raising and breeding. However, its molecular responses to copper remains largely unknown. In the present research, RNA-seq was used to mine the alteration in transcriptome of N. denticulate sinensis hepatopancreas under copper exposure. A total of 16,423 DEGs was identified between control and Cu2+ treatment groups. GO enrichment analysis of all DEGs suggested down-regulated genes exceeded up-regulated genes in all the significantly enriched terms, except for RNA polymerase III complex (GO:0005666). KEGG analysis showed Cu exposure only induced two significantly enriched pathways, including Phagosome (ko04145) and Pathogenic Escherichia coli infection (ko05130). Besides, pattern recognition receptors as Toll, lectin B, CTL1 and SRB, AMPs as crustin type I, lysozyme, and NOS were down-regulated after Cu2+ exposure, while hemocyanin, MT, HSP70 and HSP90 were significantly up-regulated, implying these molecules may play vital role in Cu2+ detoxification of N. denticulate sinensis. Our results here provide research direction of heavy metal detoxification of N. denticulate sinensis, simultaneously enriched its genomic information.

A model for phthalic acid esters' biodegradability and biotoxicity multi-effect pharmacophore and its application in molecular modification.

The objective of this study was to investigate 13 phthalic acid esters (PAEs) with medium or long straight-alkyl-chain, branching or unsaturated side chains, because their structural characteristics make them difficult to biodegrade or highly toxic. A biodegradability and biotoxicity multi-effect pharmacophore model was built using comprehensive evaluation method. The results suggested that introducing hydrophobic groups to the side chains of the PAEs could improve the molecules' biodegradability and biotoxicity effects simultaneously. Thus, 40 target PAE (HEHP, DNOP, DUP) derivatives were designed. Two environmentally friendly PAE derivatives (HEHP-Anthryl and HEHP-Naphthyl) were screened via the test of environmental friendliness and functionality. In addition, the biodegradation and biotoxicity of derivatives were found to have improved as a result of the change in van der Waals forces between molecules and their corresponding proteins. Moreover, the environmental safety of the screened PAE derivatives was confirmed by predicting the toxicity of their intermediates and calculating the energy barrier values for biodegradation and metabolic pathways. This study could provide theoretical guidance for the practical development of environmentally friendly plasticizer.

Transcriptome analysis of Neocaridina denticulate sinensis under copper exposure.

Neocaridina denticulate sinensis is a small freshwater economic shrimp, as well as excellent laboratory model for their short life cycle and easy availability. However, the response of N. denticulate sinensis to pervasive copper pollution in aquatic environments has not been deeply investigated yet. Herein, we preformed Illumina sequencing technology to mine the alterations of cephalothorax transcriptome under 2.5 µmol/L of Cu2+ after 48 h. 122,512 unigenes were assembled and 219 unigenes were identified as significantly differentially expressed genes (DEGs) between control and Cu2+ treatment groups. Functional enrichment analysis revealed that DEGs were mostly associated with immune responses and molting, such as endocytosis, Fc gamma R-mediated phagocytosis and chitin metabolic process. Seven genes were chosen for qPCR verification, and the results showed that the transcriptome sequencing data were consistent with the qPCR results. This is the first report of transcriptome information about N. denticulate sinensis. These results provided a direction for the future research of resistance to Cu2+ in this shrimp, and simultaneously enriched gene information of N. denticulate sinensis.

Evaluation of antimicrobial, antioxidant and cytotoxic properties of bioactive compounds produced from endophytic fungi of Himalayan yew ( Taxus wallichiana) in Nepal.

Background: Endophytic fungi are largely underexplored in the discovery of natural bioactive products though being rich sources of novel compounds with promising pharmaceutical potential. In this study, Taxus wallichiana, which has huge medicinal value, was investigated for its endophytic diversity and capability to produce bioactive secondary metabolites by analyzing antioxidant, antimicrobial and cytotoxic properties. Methods: The endophytes were identified by ITS-PCR using genomic DNA samples. The secondary metabolites were extracted by solvent extraction method using ethyl acetate. The antioxidant activity was analyzed by Thin Layer Chromatography, Total Phenol Content (TPC), Total Flavonoid Content (TFC) and DPPH assay, and the antimicrobial activity was analyzed by agar-well diffusion method. Brine shrimp lethality assay was used to analyze the cytotoxicity of the fungal extracts. Results: Out of 16 different Taxus trees sampled from different locations of Dhorpatan, 13 distinctive endophytic fungi were isolated and grouped into 9 different genera: Bjerkandera, Trichoderma, Preussia, Botrytis, Arthrinium, Alternaria, Cladosporium, Sporormiella and Daldinia. The ethyl acetate extracts isolated from three endophytic fungi: Alternaria alternata, Cladosporium cladosporioides and Alternaria brassicae showed significant TPC values of 204±6.144, 312.3±2.147 and 152.7±4.958µg GAE/mg of dry extract, respectively, and TFC values of 177.9±2.911, 644.1±4.202 and 96.38±3.851µg RE/mg of dry extract, respectively. Furthermore, these three extracts showed a dose dependent radical scavenging activity with IC 50 concentration of 22.85, 22.15 and 23.001 µg/ml, respectively. The extracts of C. cladosporioides and A. brassicae also showed promising antimicrobial activity against Escherichia coli, Staphylococcus aureus and Bacillus subtilis with a minimum inhibitory concentration of 250µg/ml for all bacteria. Both the samples showed cytotoxic property against shrimp nauplii with LC 50 of 104.2 and 125.9µg/ml, respectively. Conclusions: The crude fungal extracts obtained from endophytes: A. alternata, C. cladosporioides and A. brassicae upon purification and further identification of the bioactive compounds can be a fascinating source for novel pharmaceutical agents.

Antibacterial Activity of Copper Nanoparticles-Chitosan Composite against Vibrio parahaemolyticus.

The Early Mortality Syndrome (EMS) caused by Vibrio parahaemolyticus has recently resulted in a serious loss in shrimp farms in the Mekong delta, Vietnam. Here, antibacterial activity of copper nanoparticles-chitosan composite (CuCS) against V. parahaemolyticus was investigated. Copper nanoparticles were synthesized using L-ascorbic acid as a green reducing agent and chitosan as a biopolymer matrix and stabilizing agent. The physical properties of CuCS were evaluated. Next, antibacterial activity of 2.5, 5.0, 10.0 ppm CuCS against V. parahaemolyticus inoculated in a sterilized shrimp-pond water was examined. CuCS at 2.5 ppm eliminated 91.47% and 95.26% of V. parahaemolyticus after 2 and 4 h of exposure, respectively. Complete elimination was attained following 2 h of 5.0 ppm CuCS exposure. A complete elimination of V. parahaemolyticus in a real EMS-infected shrimp-pond water was also described. This study is the first to report the antibacterial activity of CuCS against V. parahaemolyticus, an important pathogen in shrimp industry in the Mekong delta, Vietnam.

Targeting the Lowest Concentration of a Toxin That Induces a Detectable Metabolic Response in Living Organisms: Time-Resolved In Vivo 2D NMR during a Concentration Ramp.

In vivo nuclear magnetic resonance (NMR) is a powerful analytical tool for probing complex biological processes inside living organisms. However, due to magnetic susceptibility broadening, which produces broad lines in one-dimensional NMR, 1H-13C two-dimensional (2D) NMR is required for metabolite monitoring in vivo. As each 2D experiment is time-consuming, often hours, this limits the temporal resolution over which in vivo processes can be monitored. Furthermore, to understand concentration-dependent responses, studies are traditionally repeated using different contaminant and toxin concentrations, which can make studies prohibitively long (potentially months). In this study, time-resolved non-uniform sampling NMR is performed in the presence of a contaminant concentration sweep. The result is that the lowest concentration that elicits a metabolic response can be rapidly detected, while the metabolic pathways impacted provide information about the toxic mode of action of the toxin. The lowest concentration of bisphenol A (BPA) that induces a response was ∼0.1 mg/L (detected in just 16 min), while changes in different metabolites suggest a complex multipathway response that leads to protein degradation at higher BPA concentrations. This proof of concept shows it is possible, on the basis of "real-time" organism responses, to identify the sublethal concentration at which a toxin impacts an organism and thus represents an essential analytical tool for the next generation of toxicity-based research and monitoring.

Ecosystem metabolism regulates seasonal bioaccumulation of metals in atyid shrimp (Neocaridina denticulata) in a tropical brackish wetland.

Natural dissolved organic matter (DOM) forms the base of aquatic food webs and is a key environmental factor that affects the bioavailability of metals for aquatic organisms. Aquatic communities are naturally exposed simultaneously to environments containing a mixture of metals and varying DOM levels and compositions. However, the exact effect of DOM on metal bioaccumulation is difficult to predict due to temporal and spatial variations in sources, production, and consumption of DOM, and to interactions between DOM and metals. Ecosystem metabolism describes the process of organic carbon production and consumption and, therefore, the trophic status of ecosystems. However, whether and how ecosystem metabolism determines the seasonality of metal bioaccumulation remains unclear. The present study used in-situ water quality sondes and discrete field samplings to establish the relationship between the seasonality of ecosystem metabolism; related environmental and limnological regulators; the metal speciation and concentration in bulk water and sediments; and their metal bioaccumulation. The target population consisted of atyid shrimp (Neocaridina denticulata) in a brackish constructed wetland in tropical Taiwan was sampled between August 2014 and November 2015. Metal bioaccumulation displayed distinct seasonal patterns that peaked in summer (Cu, Cd, Cr, Zn, Mn, and Se) or winter (Pb and Ni). The in situ production (gross primary production) and heterotrophic consumption (ecosystem respiration) of organic matter significantly decreased with increasing waterborne DOM levels in this heterotrophic wetland. Both dissolved free metals bioavailable for respiratory surfaces (As, Zn, Cu, and Cr) and insoluble metals available for dietary intake (Mn and Ni) decreased with increasing DOM, as well as with decreasing gross primary production and ecosystem respiration. Seasonal variations of metal bioaccumulation also paralleled the transition in wetland trophic status, which reflected the effect of potential qualitative changes in the wetland DOM pool. Bioaccumulation of most metals displayed strong correlations with gross primary production, ecosystem respiration, and wetland trophic status. Our findings demonstrated that ecosystem metabolism can play a key mediating role in the seasonality of metal bioaccumulation in atyid shrimp, as it links the variation and interaction between DOM level/source, the speciation/bioavailability, and the uptake efficiency for metals by aquatic organisms. This study contributes to the temporal-specific risk assessment of aquatic metal exposure in regional environmental settings. It also reveals ecosystem-specific spectra in the context of changes in climate and environment.

What can transcriptomics reveal about the phylogenetic/structural conservation, tissue localization, and possible functions of CNMamide peptides in decapod crustaceans?

Over the past several years, in silico analyses of arthropod genomes/transcriptomes have led to the identification of several previously unknown peptide families. The CNMamides are one such peptide group, having been discovered via computational analyses of the fruit fly, Drosophila melanogaster, genome; both a CNMamide precursor and receptor were identified. Recently, a CNMamide family member, VMCHFKICNLamide (disulfide bridging between the cysteine residues), was predicted via in silico mining of a crayfish, Procambarus clarkii, transcriptome, suggesting the presence of this peptide group in members of the Decapoda. Here, using publically accessible transcriptomic data, the phylogenetic/structural conservation, tissue localization, and possible functions of the CNMamide family in decapods were explored. Evidence for CNMamide precursors was found for members of each decapod infraorder for which significant sequence data are available, suggesting a ubiquitous conservation of the CNMamide family in the Decapoda. For the Penaeoidea, Caridea, Astacidea and Achelata, the isoform of CNMamide originally identified from P. clarkii appears to be ubiquitously conserved; in members of the Brachyura, VMCHFKICNMamide (disulfide bridging between the cysteine residues) is the native isoform. Interestingly, the decapod CNMamide gene appears to also have a splice variant in which the carboxy-terminal portion of the preprohormone containing the CNMamide peptide is replaced by one containing a different disulfide bridged peptide that is structurally unrelated to it; this second peptide shows considerable conservation within, but variation among, decapod infraorders. A highly conserved putative CNMamide receptor was identified from members of the Penaeoidea, Astacidea and Brachyura. Phylogenetic analyses support the annotation of the decapod receptor as a true member of the CNMamide receptor family. The presence of precursor and receptor transcripts in both nervous system- and reproductive tissue-specific transcriptomes suggests CNMamides serve as modulators of decapod neural and reproductive control systems.

Habitat selection response of the freshwater shrimp Atyaephyra desmarestii experimentally exposed to heterogeneous copper contamination scenarios.

In contaminated aquatic ecosystems, it is expected that organisms suffer some effects caused by the contaminants. However, for mobile organisms inhabiting heterogeneously contaminated ecosystems, the continuous exposure to contaminants can be avoided by moving to less contaminated habitats. The present study evaluated the habitat selection of the freshwater shrimp Atyaephyra desmarestii experimentally exposed to different copper concentrations to verify whether the heterogeneous contamination distribution and the connectivity between habitats with different copper levels could generate a random population distribution similar to metapopulation. The experiments were performed in the HeMHAS (Heterogeneous Multi-Habitat Assay System), a non-forced multi-compartmented exposure system, in which it is possible to simulate the distribution of contaminants in a linear gradient or as patches of contamination. Copper was used to simulate a linear contamination gradient (26 to 105 µg/L Cu) and two patchy scenarios with three contamination levels [reference zone (R: 26 ±â€¯7 µg/L Cu), mixing zone (M: 61 ±â€¯2 µg/L Cu) and disturbed zone (D: 101 ±â€¯12 µg/L Cu)], with two mixing zones or one central mixing zone in a heterogeneous scenario. In the copper gradient scenario, a clear trend of shrimps (59.6 ±â€¯8.0% of the population) moving to the reference zones and an avoidance of 66.7 ±â€¯11.1% of the most contaminated zone were observed. For the patchy scenarios, a random distribution of organisms (34, 36 and 30% for R, M and D zones, respectively) was observed in the scenario with one mixing zone; on the other hand, a slight preference for the reference zones (44.9 ±â€¯4.8%) was evidenced in the scenario with two mixing zones. As shrimps are able to select less contaminated areas, it is highly important to preserve clean zones in heterogeneously contaminated environments, such as the arrangement in meta-ecosystems, as the less- or uncontaminated zones might represent less stressful areas to protect populations against continuous contamination exposure.

The toxicity of non-aged and aged coated silver nanoparticles to the freshwater shrimp Paratya australiensis.

Nanoparticles (NPs) transform in the environment which result in alterations to their physicochemical properties. However, the effects of aging on the toxicity of NPs to aquatic organisms remain to be determined. Further the reports that have been published present contradictory results. The aim of this study was to examine the stability of differently coated silver nanoparticles (AgNPs) in media and the influence of aging of these NP on potential toxicity to freshwater shrimp Paratya australiensis. Coating-dependent changes in the stability of AgNP were observed with aging. Curcumin (C) coated AgNPs were stable, while tyrosine (T) coated AgNPs and epigallocatechin gallate (E) coated AgNPs aggregated in the P. australiensis medium. Increased lipid peroxidation and catalase activity was noted in P. australiensis exposed to AgNPs, suggesting oxidative stress was associated with NP exposure. The enhanced oxidative stress initiated by aged C-AgNPs suggests that aging of these NPs produced different toxicological responses. In summary, data suggest that coating-dependent alterations in NPs, together with aging affect both persistence and subsequent toxicity of NPs to freshwater organisms. Thus, the coating-dependent fate and toxicity of AgNPs together with the effect of their aging need to be considered in assessing the environmental risk of AgNPs to aquatic organisms.

Avoidance response by shrimps to a copper gradient: Does high population density prevent avoidance of contamination?

Bioassays using the nonforced exposure approach have been shown to be a relevant tool that might complement the traditional ecotoxicological risk assessment. Because the nonforced exposure approach is based on spatial displacement of organisms and the consequent habitat selection processes, the population density might play an important role in the decision to avoid or prefer an ecosystem. Therefore, the aim of the present study was to assess if the avoidance response to contamination, measured in a nonforced exposure system, is density-dependent and how determinant contamination could be for the habitat-selection process in comparison with the population density. The freshwater shrimp Atyaephyra desmarestii was exposed to a copper gradient in a nonforced exposure system formed by 7 interconnected compartments (total volume 600 mL), which contained different copper concentrations. The density treatments used were 3, 5, and 10 organisms per compartment, corresponding to 0.5, 0.8, and 1.7 organisms per 100 mL, respectively. Clearly, the avoidance response to copper was more intense in the population with the lower density: the highest population density showed the lowest avoidance. The concentrations that triggered an avoidance of 50% of the population were 47, 134, and 163 µg L-1 . In summary, it was observed that shrimps were able to detect and avoid potentially toxic copper concentrations but that the avoidance response was affected by population density. Environ Toxicol Chem 2018;37:3095-3101. © 2018 SETAC.

Do trace metal contamination and parasitism influence the activities of the bioturbating mud shrimp Upogebia cf. pusilla?

Mud shrimp are considered as among the most influential ecosystem engineers in marine soft bottom environments because of their significant bioturbation activity and their high density. These organisms play a key role on the physical structure of sediments through intense sediment reworking activity and also deeply influence geochemical properties of sediments via frequent bioirrigation events. The influence that mud shrimp have on the environment is related to the magnitude of bioturbation processes and subsequently depends on their physiological condition. In natural environments, several factors act together and influence the well-being of organisms. Among them, the deleterious role of parasites on the physiology and the behavior of their host is well established. Aquatic organisms are also subject to pollutants released by anthropogenic activities. However, the effect of both stressors on the fitness and bioturbation activity of mud shrimp has never been investigated yet. We conducted a 14-day ex-situ experiment to evaluate the influence of trace metal contamination (cadmium Cd) and parasitism infestation on the gene expression (molecular endpoint) and sediment reworking activity (behavioral endpoint) of the mud shrimp Upogebia cf. pusilla. At completion, mud shrimp exhibited substantial Cd bioaccumulation, with parasitized organisms showing a significantly lower contaminant burden than unparasitized specimens. Cadmium contamination induces modifications of gene expression in both unparasitized and parasitized organisms. We report an antagonistic effect of both stressors on gene expression, which cannot be fully explained by a lower Cd bioaccumulation. At the behaviour level, parasitism seems to reduce the sediment reworking activity of mud shrimp, while Cd contamination appears to stimulate this activity. This study highlights that the effects of multiple stressors may be quite different from the effects of each stressor considered individually. It should also motivate for more studies evaluating the influence of multiple stressors on different endpoints encompassing various levels of organization.

Spatial occurrence and effects of microplastic ingestion on the deep-water shrimp Aristeus antennatus.

Microplastic (MP) ingestion has been reported in a wide variety of organisms, however, its spatial occurrence and effects on wild populations remain quite unknown. The present study targets an economically and ecologically key species in the Mediterranean Sea, the shrimp Aristeus antennatus. 39.2% of the individuals sampled had MP in their stomachs, albeit in areas close to Barcelona city the percentage reached values of 100%. Overall, MP ingestion was confirmed in a wide spatial and depth (630-1870 m) range, pointing out the great dispersion of this pollutant. The benthophagous diet and close relationship with the sea bottom of A. antennatus might enhance MP exposure and ultimately lead to accidental ingestion. Detailed analysis of shrimps' diet revealed that individuals with MP had a higher presence of endobenthic prey. Microplastic fibers are probably retained for long periods due to stomach's morphology, but no negative effects on shrimp's biological condition were observed.

Evaluating the non-lethal effects of organophosphorous and carbamate insecticides on the yabby (Cherax destructor) using cholinesterase (AChE, BChE), Glutathione S-Transferase and ATPase as biomarkers.

The toxicity of two organophosphorus insecticides, chlorpyrifos (CPF), malathion (MAL), and one carbamate insecticide, methomyl (METH), to the yabby (Cherax destructor) was assessed by measuring cholinesterase (AChE, BChE), Glutathione S-Transferase (GST) and Na+/K+ATPase activity after 96h of exposure. Yabbies exposed to all three insecticides at 2 and 5µgL-1 exhibited significant AChE, BChE, GST and Na+/K+ATPase inhibition. Based on these enzyme inhibition tests, the toxicity of the three insecticides to C. destructor was CPF > MAL > METH. After 14 days of recovery the yabbies enzymatic activities of AChE, BChE, GST and Na+/K+ATPase was measured. Recovery of The enzyme activity recovery was faster after the exposure to METH than for the yabbies exposed to CPF and MAL. Slow recovery of enzyme activity could affect the physical activities of organisms and produce indirect effects on populations if such crayfish are less able to elude predators or search for food.

Acute and sub-chronic effects of sub-lethal cadmium exposure on energy metabolism in the freshwater shrimp, Paratya curvirostris.

Cadmium (Cd) is a toxic trace element enriched in waters through activities such as mining and agriculture. The freshwater shrimp Paratya curvirostris inhabits near-coastal, lowland streams potentially impacted by Cd, but nothing is known regarding its sensitivity to this metal. An acute (96h) median lethal concentration (LC50) of 405µgL-1 was derived for P. curvirostris, placing it among the most tolerant of freshwater shrimp species. Acute (4 d; 0, 50 and 100µgL-1) and sub-chronic (10 d; 0, 25 and 50µgL-1) exposures then investigated effects of Cd on energy metabolism (respiration rate, excretion rate, O:N ratio). In contrast to effects in previously studied species, Cd induced an increased respiration rate, which when coupled with an unchanged excretion rate, resulted in an increased O:N ratio. These data were explained by an increased reliance on carbohydrate and/or lipid as a metabolic substrate stimulated by increased metabolic costs of toxicant exposure. Similar effects were seen across all time-points, although the lowest effective Cd concentration decreased with increased exposure time. Overall, results suggest that Cd is unlikely to be a significant environmental stressor to P. curvirostris, except in highly contaminated freshwaters, and/or where Cd co-occurs with hypoxia.

De novo assembly and analysis of changes in the protein-coding transcriptome of the freshwater shrimp Paratya australiensis (Decapoda: Atyidae) in response to acid sulfate drainage water.

BACKGROUND: The atyid shrimp Paratya australiensis occurs in surface freshwater habitats throughout eastern Australia and has been used to study the ecotoxicology of contaminants such as pesticides and metals. The acidification of surface water that can occur after acid sulfate material in soils and sediments is oxidised and subsequently re-wetted is a serious environmental issue in coastal regions and inland riverine floodplains worldwide. Solubilisation of soil-associated minerals can result in high waterborne concentrations of mineral salts and dissolved metals, which together with low pH represent a potential threat to aquatic ecosystems in affected regions. The aims of the present study were to gain insight into stress responses induced by exposure to acid drainage water (ADW) in P. australiensis by determining changes in the abundance of protein-coding transcripts and to generate a comprehensive transcriptomic resource to facilitate further research into gene regulation or protein structure and function in this species. Adult P. australiensis were exposed for 24 h to undiluted ADW, 50 % ADW diluted in river water, or to river water as control, and high-throughput mRNA sequencing (RNA-Seq) conducted on whole-body tissues. A reference transcriptome was generated using de novo assembly and putative protein-coding regions were identified and annotated. Changes in transcript abundance in response to ADW exposure were determined by aligning reads to the reference transcriptome and quantifying coverage. RESULTS: A high proportion of arthropod benchmarking universal single-copy orthologues were present in the reference transcriptome. Functions associated with cuticle biosynthesis and oxidative stress were significantly enriched in the lists of transcripts exhibiting differential abundance in either direction after exposure to 50 % or 100 % ADW. Transcripts involved in osmoregulation exhibited decreased abundance following exposure to ADW. The transcriptome contained full-length coding sequences for numerous proteins known to be involved in environmental response pathways, including two putative metallothioneins, four glutathione peroxidases and 19 nuclear receptors. CONCLUSIONS: The results of the present study provide insight into stress response pathways induced in crustaceans by short-term exposure to multiple stressors present in ADW such as low pH, high salinity and dissolved metals, and represent a resource for future toxicogenomics and protein functional studies in P. australiensis.

Microencapsulation of norfloxacin in chitosan/chitosan oligosaccharides and its application in shrimp culture.

Norfloxacin chitosan/chitosan oligosaccharide microcapsules (NCCM) were prepared by emulsion-chemical crosslinking method. The characteristics of obtained microcapsules were evaluated by scanning electron microscopy, Fourier transform infrared spectroscopy and release experiments. Cumulative release profile of norfloxacin from the chitosan microcapsules in natural seawater was measured and the controlled release of drugs was at a uniform rate in 48h. The chitosan microcapsules were applied onto the antibacterial study of the shrimp culture in natural seawater. It is observed that the seawater in the NCCM added groups was relatively clear and the biomass of Vibrio increased slowly in contrast to the control and norfloxacin groups. The inhibition rate of Vibrio in norfloxacin groups obvioursly decreased after the 5th day, whereas, it remained high and stable during experiment period in NCCM groups. The results showed that the chitosan microcapsules as release materials have excellent antibacterial effects on Vibrio in the farming of Penaeus vannamei Boone. The controlled release could obviously reduce dosage of antibiotics and delivery times, and effectively improve the utilization rate of norfloxacin drugs for shrimps.

Influence of temperature on toxicity of single pharmaceuticals and mixtures, in the crustacean A. desmarestii.

Lethal and sublethal responses of the shrimp Atyaephyra desmarestii exposed to three pharmaceutical compounds, Diclofenac (DF), Ibuprofen (IB) and Carbamazepine (CBZ), individually and in mixtures, were evaluated under two temperature scenarios. LC50 (96h) values were obtained individually at 20° and 25°C. At 25°C, mortality in binary and ternary mixtures is higher than at 20°C. The toxicity of the mixtures was predicted on the basis of individual mortality data using two toxicity models: Concentration addition (CA) and Independent action (IA). Our results showed that neither CA nor IA unequivocally predicted the observed toxicity of binary and ternary mixtures. For sublethal toxicity, selected endpoints were: ingestion rate, osmoregulatory capacity and respiration rate. Regarding osmoregulatory capacity, no significant differences were found. The highest ingestion rates were recorded in organisms exposed at 25°C, irrespective of the compound, after 30 and 60min of exposure. At 20°C, there was a significant decrease in respiration rate (Dunnett́s test p<0.05) under conditions of severe anoxia (1mg O2L(-1)) in organisms exposed to 13.3µgL(-1) of DF. At 25°C a significantly lower respiration rate with respect to the control (Dunnett́s test p<0.05) was found in organisms exposed to 13.8µgL(-1) of CBZ under conditions of moderate hypoxia and well-oxygenated water (3 and 5mg O2L(-1), respectively). The respiratory independence of organisms exposed to the higher temperature (25°C) also decreased. This study shows that CBZ and DF individually, even at relatively low concentrations, may produce respiratory deficiencies in the freshwater shrimp, Atyaephyra desmarestii under certain temperature and water oxygenation conditions, thus reducing its ability to function.

Development of an ecotoxicological protocol for the deep-sea fauna using the hydrothermal vent shrimp Rimicaris exoculata.

In light of deep-sea mining industry development, particularly interested in massive-sulphide deposits enriched in metals with high commercial value, efforts are increasing to better understand potential environmental impacts to local fauna. The aim of this study was to assess the natural background levels of biomarkers in the hydrothermal vent shrimp Rimicaris exoculata and their responses to copper exposure at in situ pressure (30MPa) as well as the effects of depressurization and pressurization of the high-pressure aquarium IPOCAMP. R. exoculata were collected from the chimney walls of the hydrothermal vent site TAG (Mid Atlantic Ridge) at 3630m depth during the BICOSE cruise in 2014. Tissue metal accumulation was quantified in different tissues (gills, hepatopancreas and muscle) and a battery of biomarkers was measured: metal exposure (metallothioneins), oxidative stress (catalase, superoxide dismutase, glutathione-S-transferase and glutathione peroxidase) and oxidative damage (lipid peroxidation). Data show a higher concentration of Cu in the hepatopancreas and a slight increase in the gills after incubations (for both exposed groups). Significant induction of metallothioneins was observed in the gills of shrimps exposed to 4µM of Cu compared to the control group. Moreover, activities of enzymes were detected for the in situ group, showing a background protection against metal toxicity. Results suggest that the proposed method, including a physiologically critical step of pressurizing and depressurizing the test chamber to enable the seawater exchange during exposure to contaminants, is not affecting metal accumulation and biomarkers response and may prove a useful method to assess toxicity of contaminants in deep-sea species.

Physiological effects of essential metals on two detritivores: Atyaephyra desmarestii (Millet) and Echinogammarus meridionalis (Pinkster).

Freshwater ecosystems are essential for humans; however, input of several types of contamination has led to the degradation of these ecosystems. Thus, it is urgent to assess their health to allow actions for prevention and remediation. The level of trace metals can be enhanced by natural or anthropogenic sources. Essential metals, such as copper and zinc, become toxic when present in the environment above threshold concentrations. To evaluate the physiological effects of these 2 essential metals for 2 freshwater detritivores, the shrimp Atyaephyra desmarestii and the amphipod Echinogammarus meridionalis, acute tests were performed. Forty-eight hour median lethal concentration (LC50) values were estimated for these species using static bioassays with copper and zinc. Sublethal assays for both metals with several phases were also done to evaluate the effects on feeding behavior. The LC50 values of copper for the shrimp A. desmarestii and amphipod E. meridionalis were 0.128 mg/L and 0.050 mg/L and those of zinc were 7.951 mg/L and 11.860 mg/L, respectively. The results indicated that copper is more toxic to both species. Only E. meridionalis showed deleterious effects of copper on feeding rate. Zinc showed some tendency for feeding inhibition in both species. Environ Toxicol Chem 2016;35:1442-1448. © 2015 SETAC.