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The metallurgy industry is a potent global source of particulate matter (PM) atmospheric emissions. A portion of this PM may settle in aquatic (SePM) carrying metal/metalloid particles and metallic nanoparticles. Surprisingly, this form of contamination has not received due attention from most environmental monitoring agencies. We analyzed the effect of exposure to SePM on shrimp post-larvae, a critical stage for the viability of shrimp populations and for the trophic chain. After acclimation, shrimp were exposed to contaminants using a randomized experimental design-a 4 × 4 factorial arrangement with 2 factors: exposure time (24, 48, 72, and 96 h) and SePM concentration (0.00, 0.01, 0.10, and 1.00 g L -1 ). The bioaccumulation of metals, contamination rates, mortality, and ROS-related biomarkers (lipid peroxidation - LPO; DNA strand breakage DNA SB and metallothionein content - MTs;) were evaluated. After contamination, the water contained 27 different metals/metalloids. Post-larvae accumulated metals, such as Cd, Pb, Al, As, Se, Sr, Zr, Ba, La, Ce, W, and Hg. However, the rise in SePM did not result in a proportional bioaccumulation rise, indicating that effective biological barriers may work for some metals. Although the different levels of SePM changed mortality dynamics, they resulted in a similar final lethality (60-80 %). SePM caused significant damage to lipids (increased LPO), genetic material (DNA SB), and increased Mts. Such effects may reflect a particularly deleterious ecological problem as it is present at such an early stage of life. These results identified a clear environmental risk since the lower level of exposure used was 10 2 times lower than that measured in the habitats affected by local industry. Consequently, our results emphasize the need for clear protocols for monitoring the effects of SePM in aquatic environments., 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 © 2024 Elsevier B.V. All rights reserved.)

Nanoplastics (NPs) are widely distributed in the aquatic environment and have become a global concern as a new type of pollutant. Many researchers have studied the physiological effects of NPs on aquatic organisms, but relatively little is known about their effects on intestinal immune function in crustaceans. Therefore, we used NPs concentrations of 0, 5, 10, 20, 40 mg/L for 28 days of stress, evaluated the effects of NPs exposure on intestinal cell apoptosis, histopathological damage, and glutathione (GSH) metabolism of juvenile East Asian river prawns (Macrobrachium nipponense). As NPs concentration increased, the contents of total GSH and oxidized glutathione decreased gradually (P < 0.05), the concentration of GSH first increased and then decreased (P < 0.05), and the activities of lysozyme, acid phosphatase, phenoloxidase, and alkaline phosphatase first increased and then decreased (P < 0.05). Additionally, intestinal tissue structure was damaged, and the apoptosis rate significantly increased (P < 0.05). The expression of intestinal autophagy genes (CTL, ALF, Crustin, ATG8, and BCL-2) increased at first and then decreased, the expression levels of TNF and Wnt4 significantly decreased, and the expression of Beclin significantly increased with increasing NPs concentration. We also found that AP-1 and PTEN were highly expressed in the hepatopancreas and were involved in intestinal immune responses. Our results showed that exposure to NPs may induce apoptosis of intestinal tissue cells, induce autophagy, and inhibit GSH metabolism, thereby reducing intestinal immune function of M. nipponense. These findings provide a reference for healthy aquaculture and ecological risk assessment of prawns., Competing Interests: Declaration of competing interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled, “Polystyrene nanoplastics induce apoptosis, histopathological damage, and glutathione metabolism disorder in the intestine of juvenile East Asian river prawns (Macrobrachium nipponense)”., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This work studied the effects of C. longa extract as a dietary supplement for Macrobrachium rosenbergii. Shrimp juvenile (n = 960, initial average weight 0.06 ± 0.042 g and initial stocking density 0.4 animals/L) received diets with different concentrations of turmeric extract (0.05, 0.2, 1%) for 60 days, and zootechnical (weight gain, specific growth rate, initial and final condition factor) and metabolic (activities of digestive enzymes, amino acid metabolism enzymes, and energy metabolism enzymes) parameters were evaluated. A transport simulation (TS) test was conducted and, in challenged animals, survival and antioxidant status were studied. Phenolic compound content and in vitro antioxidant potential of the turmeric extract were evaluated. Turmeric extract (0.05%) significantly increased weight gain and the activity of digestive enzymes. An increase in citrate synthase activity for all concentrations of turmeric was also observed. The glutamate dehydrogenase activity in the 0.05% group was also increased by turmeric extract. The in vitro antioxidant activity of the turmeric extract was like butylated hydroxytoluene (BHT), a common antioxidant, and the total phenolic content was 0.98 ± 0.02 mmolAG/g. In animals from the TS test, turmeric extract promoted a dose-dependent increase in animal survival. Furthermore, turmeric supplementation increased the activity of the enzymes catalase, glutathione reductase, glutathione peroxidase, glutathione S-transferase, and it reduced glutathione (GSH) content. A reduction in the levels of lipid peroxidation (TBARS) and protein carbonyl was also observed in the hepatopancreas of all experimental groups. The addition of low concentrations of turmeric extract to the diet of M. rosenbergii juvenile presents great potential for application in commercial shrimp farming, since the extract (at concentrations as low as 0.05 to 1%) increased weight gain and improved the physiological and metabolic conditions of the animals, improving the use of nutrients and consequently the animals' responses to stressful conditions, such as transport., Competing Interests: Declarations. Ethical approval: This declaration is not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study investigated the effects of Bisphenol A (BPA), a common endocrine-disrupting chemical, on the antioxidant enzyme activities in the hepatopancreas and the expression of genes related to ovarian development in oriental river prawns (Macrobrachium nipponense). The 24hLC50 and 48hLC50 values for BPA were 80.59 mg/L and 63.90 mg/L, respectively, with a safe concentration of 12.06 mg/L. Prawns were exposed to low (4.85 mg/L), safe (12.06 mg/L), and high (30.00 mg/L) concentrations of BPA for 10 days to measure enzyme activities, and for 20 days followed by 7 days in BPA-free water to measure gene expression. Short-term exposure (12 h, 1d, 3d) to low concentration BPA did not significantly affect superoxide dismutase (SOD) activity in the hepatopancreas (P > 0.05), but long-term exposure (6d, 10d) significantly reduced SOD activity (P < 0.05). Catalase (CAT) activity showed no significant changes throughout the low concentration exposure period (P > 0.05). At safe and high concentrations, SOD and CAT activities significantly decreased after 12 h of exposure (P < 0.05). BPA affected heat shock protein 90 (HSP90) expression in the ovary, with low concentration BPA significantly upregulating HSP90 after 1 day (P < 0.05), but returning to normal levels after 10 and 20 days. At the safe concentration, HSP90 was significantly upregulated at all three sampling points (1d, 10d, 20d) (P < 0.05), while high concentration exposure led to significant upregulation only on day 10 (P < 0.05). Low concentration BPA had no significant effect on Cathepsin B (CB) and Cathepsin L (CL) gene expression in the ovaries (P > 0.05). However, safe concentration exposure promoted CB expression on days 1, 10, and 20 (P < 0.05), while high concentration exposure significantly increased CB expression on day 1 (P < 0.05), with levels returning to normal on days 10 and 20. CL expression significantly increased after 20 days of exposure to both safe and high concentrations (P < 0.05). Gene expression levels in the ovaries returned to normal after transfer to BPA-free water, with HSP90 and CB normalizing by day 1, and CL by day 7. These results indicate that even safe concentrations of BPA impose stress on the hepatopancreas and increase the expression of HSP90, CB, and CL genes in the ovaries, affecting ovarian development. And, these effects are reversible within a certain period after the removal of BPA., Competing Interests: Declaration of competing interest The authors declare that there are no known competing financial interests or personal relationships that could have appeared to influence the work reported in this study., (Copyright © 2024. Published by Elsevier B.V.)

The aim of this study was to evaluate the sensitivity of the prawn Palaemon argentinus to the pyrethroid cypermethrin (CYP) and the tetramic acid spirotetramat (STM). These treatments were compared with prawns collected at a reference site to define their basal physiological state. Initially, physicochemical parameters and several pollutants at the selected site were analyzed. The LC 50 -96 h was determined in adult prawns. Then, prawns were exposed for 96 h to sublethal concentrations of CYP (0.0005 μg/l) and STM (0.44 mg/l) to evaluate the effects on some biochemical endpoints. A treatment combining both pesticides was also added at 5 % of these values. Controls with and without solvent (acetone) were included. The LC 50 -96 h values were 0.005 μg/l and 4.43 mg/l for CYP and STM, respectively. Moreover, some biomarkers linked to oxidative and energy metabolism were analyzed in the hepatopancreas and muscle of both essayed prawns and those at the basal state. The STM caused a significant decrease in total protein content (32 %) in contrast to the increase of protein carbonyl content (71 %) (p < 0.05). Also, glutathione S-transferase (52 %) and catalase (61 %) activities in the hepatopancreas of exposed prawns were higher compared to both the control and state basal groups (p < 0.05). In muscle, only a significant decrease in the lactate content (69 %) was caused by STM (p < 0.05). In addition, CYP caused a significant increase in the lactate dehydrogenase activity (110 %) in muscle and triacylglycerol content (73 %) in the hepatopancreas (p < 0.05). The integrated biomarker index (IBRv2) analysis showed that STM caused greater damage than CYP. Besides, the combined treatment showed an antagonistic interaction between both insecticides. The differential response of biomarkers to both CYP and STM exposure with respect to their basal levels shows a high sensitivity of P. argentinus demonstrating its potential role as a bioindicator organism., 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 © 2024 Elsevier B.V. All rights reserved.)

Microplastics (MPs) have unique toxicokinetic (TK) processes that differ from those of soluble pollutants. This study investigated the ingestion, migration, accumulation, and clearance of environmental aging MPs in the Japanese swamp shrimp (Macrobrachium nipponense). The concentrations of plastic additives and personal care products adsorbed onto MPs in natural river water were determined, and TK models for MPs and MPs-loaded pollutants were developed. Results showed that the formation of surface biofilms and alterations in the distribution of MPs in waters caused by environmental aging affect MPs bioavailability, which is mainly related to the feeding habits of shrimp. The decrease in MPs particle size caused by biological digestion and the increase in the number of oxygen-containing functional groups caused by environmental aging affect the TK process of MPs. The TK model of MPs-loaded pollutants revealed the cleaning effect of shrimp on pollutants adsorbed onto MPs during swallowing and spitting MPs. This cleaning effect significantly increases the bioavailability of MPs-associated pollutants in aquatic environments. This study provides a new perspective for understanding the interactions between environmental MPs and their associated pollutants in aquatic ecosystems., 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 © 2024 Elsevier B.V. All rights reserved.)

In recent years, there has been extensive research within the scientific community on deep eutectic systems due to their remarkable versatility in solubilizing diverse substances and serving as effective solvents in catalytic processes. While initially regarded as non-toxic, a comprehensive toxicological assessment is essential to comprehend their behavior within organisms. In this study, seven distinct systems, composed of N,N,N-triethyl-N-(2,3-dihydroxypropyl)ammonium chloride (N00Cl) and glycerol-derived ethers with alkyl chains of varying lengths (100, 200, 3F00, 300, 3i00, and 400), in a 1:2 molar ratio were investigated for their aquatic toxicity in shrimp (Palaemon varians). The assessment involved analyzing oxidative stress biomarkers such as glutathione S-transferase, glutathione peroxidase, catalase, superoxide dismutase, total antioxidant capacity (TAC), and lipoperoxidation (MDA content). Results show an odd-even effect for LC 50 values being N00Cl-300, the system showing higher values. Regarding oxidative stress, an imbalance between reactive oxygen species (ROS) and antioxidant capacity in the organisms has been observed, suggesting significant toxicity to shrimps due to the changes in oxidative stress biomarkers at high concentrations. However, at 100 mg/l all systems can be considered environmentally safe, and no negative impacts are expected on aquatic ecosystems., (© 2024. The Author(s).)

Habitat selection by aquatic organisms is dependent on the availability of adequate conditions to support life and the benefits that the habitat provides. Contaminated environments tend to be less attractive to organisms because reduced habitat quality leads to increased maintenance costs. Consequently, reduced colonization of such disturbed habitats is an expected response. However, colonization has been understudied as an ecotoxicological test endpoint, despite its proven ability to assess habitat selection by populations across various taxa. The aim of the present study was to investigate whether previous exposure to copper could alter the colonization behavior of the freshwater prawn Macrobrachium rosenbergii along a non-forced exposure gradient of interconnected copper-contaminated habitats (0 to 500 µg/L) due to physiological and behavioral impairments. To assess this, post-larvae of M. rosenbergii were pre-exposed to 0, 50, 250 and 500 µg/L copper for a maximum period of 48 h. The physiological status and motility of the organisms after pre-exposure to copper were evaluated using behavioral endpoints (swimming activity by video tracking) and biochemical biomarkers (biotransformation, oxidative stress and neurotoxicity). The results indicated that pre-exposure to copper (at concentrations of 0, 50 and 500 µg/L) significantly influenced the median colonization concentration (CC50), which decreased from 270 µg/L to 109 µg/L. None of the assessed swimming parameters (speed, motility rate, exploration rate, and total distance) were affected by the pre-exposure to copper (0, 50 and 250 µg/L). Biochemically, cholinesterase levels were only affected in the prawn population exposed to 250 µg/L of copper. The present study provides a better understanding of the relevance of colonization as an ecotoxicological endpoint for assessing the spatial distribution of populations, including both new inhabitants and previously exposed organisms, in recovering habitats., 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 © 2024 Elsevier B.V. All rights reserved.)

Macrobrachium nipponense is an important commercial freshwater species in China. However, the ability of alkali tolerance of M. nipponense is insufficient to culture in the major saline-alkali water source in China. Thus, it is urgently needed to perform the genetic improvement of alkali tolerance in this species. In the present study, we aimed to analyse the effects of alkali treatment on gills in this species after 96 h alkalinity exposure under the alkali concentrations of 0 mmol/L, 4 mmol/L, 8 mmol/L, and 12 mmol/L through performing the histological observations, measurement of antioxidant enzymes, metabolic profiling analysis, and transcriptome profiling analysis. The results of the present study revealed that alkali treatment stimulated the contents of malondialdehyde, glutathione, glutathione peroxidase in gills, indicating these antioxidant enzymes plays essential roles in the protection of body from the damage, caused by the alkali treatment. In addition, high concentration of alkali treatment (> 8 mmol/L) resulted in the damage of gill membrane and haemolymph vessel, affecting the normal respiratory function of gill. Metabolic profiling analysis revealed that Metabolic pathways, Biosynthesis of secondary metabolites, Biosynthesis of plant secondary metabolites, Microbial metabolism in diverse environments, Biosynthesis of amino acids were identified as the main enriched metabolic pathways of differentially expressed metabolites, which are consistent with the previous publications, treated by the various environmental factors. Transcriptome profiling analyses revealed that the alkali concentration of 12 mmol/L has more regulatory effects on the changes of gene expression than the other alkali concentrations. KEGG analysis revealed that Phagosome, Lysosome, Glycolysis/Gluconeogenesis, Purine Metabolism, Amino sugar and nucleotide sugar metabolism, and Endocytosis were identified as the main enriched metabolic pathways in the present study, predicting these metabolic pathways may be involved in the adaption of alkali treatment in M. nipponense. Phagosome, Lysosome, Purine Metabolism, and Endocytosis are immune-related metabolic pathways, while Glycolysis/Gluconeogenesis, and Amino sugar and nucleotide sugar metabolism are energy metabolism-related metabolic pathways. Quantitative PCR analyses of differentially expressed genes (DEGs) verified the accuracy of the RNA-Seq. Alkali treatment significantly stimulated the expressions of DEGs from the metabolic pathways of Phagosome and Lysosome, suggesting Phagosome and Lysosome play essential roles in the regulation of alkali tolerance in this species, as well as the genes from these metabolic pathways. The present study identified the effects of alkali treatment on gills, providing valuable evidences for the genetic improvement of alkali tolerance in M. nipponense., (© 2024. The Author(s).)

Bicarbonate and sulfate are among two primary ion constituents of saline-alkaline water, with excessive levels potentially causing metabolic disorders in crustaceans, affecting their molting and interrupting development. As an economically important crustacean species, the molecular adaptive mechanism of giant freshwater prawn Macrobrachium rosenbergii in response to the stress of bicarbonate and sulfate remains unexplored. To investigate the mechanism underlying NaHCO 3 , Na 2 SO 4 , and mixed NaHCO 3 , Na 2 SO 4 stresses, M. rosenbergii larvae were exposed to the above three stress conditions, followed by total RNA extraction and high-throughput sequencing at eight distinct time points (0, 4, 8, 12, 24, 48, 72, and 96 h). Subsequent analysis revealed 13, 16, and 13 consistently identified differentially expressed genes (DEGs) across eight time points under three stress conditions. These consistently identified DEGs were significantly involved in the Gene Ontology (GO) terms of chitin-based cuticle development, protein-carbohydrate complex, structural constituent of cuticle, carnitine biosynthetic process, extracellular matrix, and polysaccharide catabolic process, indicating that alkaline stresses might potentially impact the energy metabolism, growth, and molting of M. rosenbergii larvae. Particularly, the transcriptome data revealed that DEGs associated with energy metabolism, immunity, and amino acid metabolism were enriched across multiple time points under three stress conditions. These DEGs are linked to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including glycolysis/glucogenesis, amino sugar and nucleotide sugar metabolism, and lysine degradation. Consistent enrichment findings across the three stress conditions support conclusions above. Together, these insights are instrumental in enhancing our understanding of the molecular mechanisms underlying the alkaline response in M. rosenbergii larvae. Additionally, they offer valuable perspectives on the regulatory mechanisms of freshwater crustaceans amid saline-alkaline water development., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Banana (Musa acuminata) blossom contains high nutritional value and bioactive compounds. In this study, Macrobrachium rosenbergii were fed with diets containing banana blossom powder (BBP) at 10 and 20 g kg -1 , hot-banana blossom (BBH) extract at 10 and 20 g kg -1 , and the basal diet for 56 days. The growth performance, physiological response and immune parameters were evaluated. The results showed that a significantly higher percentage weight gain (PWG) and percentage length gain (PLG) in prawns fed with BBH diet. The feed efficiency (FE) significantly increased in prawns fed BBP. The prawn fed both BBH and BBP diet showed higher survival rate than control group. The prawn fed with BBH showed a significant increase in total haemocyte count (THC) and different haemocyte count (DHC), whereas phenoloxidase (PO) activity and respiratory bursts (RBs) significant increase in prawns fed both BBP and BBH diet. Furthermore, M. rosenbergii fed with both BBP and BBH diets showed significantly higher phagocytic activity and clearance efficiency against Lactococcus garvieae infection. At the end of the 56 days of feeding trial, the susceptibility of prawns to L. garvieae infection and hypothermal (18 °C) stress were evaluated. The results showed that prawns fed BBH diets had a significantly higher survival rate against L. garvieae than those of fed with the basal diet. Anti-hypothermal stress was observed in prawns fed both BBP and BBH diets showing no significant difference in haemolymph glucose in prawns subjected to 18 °C and 28 °C, whereas the norepinephrine level in haemolymph of prawns fed with BBH diets subjected to 18 °C was significantly lower than in prawns subjected to 28 °C. In summary, we recommend addition of hot-banana blossom extract to the diet of M. rosenbergii at 20 g kg -1 to promote growth performance, improve physiological function, enhance immunity, increase anti-hypothermal stress, and to increase resistance against L. gavieae., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Heavy metal Cadmium (Cd 2+ ) pollution has become a severe environmental problem for aquatic organisms. In crustaceans, gills (Gi) and hepatopancreas (Hp) play a vital role in the toxicology. However, in Macrobrachium rosenbergill, there are few researches about gill and hepatopancreases responding to Cd 2+ stress at a molecular level. In this study, transcriptomic analysis was applied to characterize gene expression profiles of gills and hepatopancreas of M. rosenbergill after Cd 2+ exposure for 0 h, 3 h and 3 d. Six cDNA libraries (Gi 0 h, Gi 3 h, Gi 3 d, Hp 0 h, Hp 3 h, and Hp 3 d) were constructed and a total of 66,676 transcripts and 48,991 unigenes were annotated. Furthermore, differentially expressed genes (DEGs) were isolated by comparing the Cd 2+ treated time-point libraries (3 h and 3 d group) with the control library (0 h group). The results showed that most of the DEGs were down-regulated after Cd 2+ exposure and the number of DEGs among gill groups were significantly higher than those among hepatopancreas groups. GO functional and KEGG pathway analysis suggested many key DEGs in response to the Cd 2+ stress, such as metallothionein and Hemocyanin. Additionally, a total of six DEGs were randomly selected to further identify their expressional profile by qPCR. The results indicated that these DEGs were involved in the response to Cd 2+ . This comparative transcriptome provides valuable molecular information on the mechanisms of responding to Cd 2+ stress in M. rosenbergii, which lays the foundation for further understanding of heavy metal stress., (© 2021. The Author(s).)

Deltamethrin (Del), a commonly used broad-spectrum insecticide, has been reported to have a toxic effect on aquatic animals, but knowledge in freshwater prawns is limited. This study revealed that Del is highly toxic to Macrobrachium nipponens with the 24 h, 48 h, 72 h, and 96 h LC 50 values to be 0.268, 0.165, 0.104, and 0.066 μg/L, respectively. To further investigate the toxic effect of Del in M. nipponense and the reversibility of damage, prawns were exposed to 0.05 μg/L Del for four days and then transferred into fresh water for seven days. Histopathological examination, oxidative stress, hepatopancreas function, respiration system, and immune system were analyzed through multiple biomarkers. Results showed that Del exposure caused severe histopathological damage to hepatopancreas and gill in M. nipponense, and the prominent decrease of acid phosphatase (ACP) and alkaline phosphatase (AKP) activity further enhanced the hepatopancreas damage; the accumulation of malonaldehyde (MDA) and hydrogen peroxide (H 2 O 2 ), and the decrease of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity, indicated severe oxidative stress caused by Del. Besides, Del exposure also induced remarkably increased lactic acid (LD) level, decreased lactate dehydrogenase (LDH) activity, and decreased expression of immune-related genes, which demonstrated the respiration disruption and immunosuppression caused by Del. After 7-day decontamination in freshwater, the indicator of hepatopancreas function (ACP and AKP activity) and respiration (LD level and LDH activity) improved to the control group level. However, the histopathological damage and the biomarker in oxidative stress and immune system did not recover to the initial level., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Deltamethrin (DM) is a synthetic pyrethroid used for agricultural purposes to control insects. However, its extensive use contaminates the aquatic environment and results in serious health problems in aquatic organisms. Knowledge about the toxic effect of DM in freshwater prawns is limited; therefore, this study aims to assess the toxicity of DM in Macrobrachium rosenbergii based on multiple biomarkers. Four-day acute toxicity tests showed that DM was highly toxic to M. rosenbergii with the 24 h, 48 h, 72 h and 96 h LC 50 values to be 1.919, 0.603, 0.539, and 0.449 μg/L, respectively. According to 96 h LC 50 , prawns were exposed to DM at three concentrations (0.02, 0.08, and 0.32 μg/L) for 4 days, and then moved into fresh water for decontamination to investigate the toxic effect of DM in M. rosenbergii. At low concentration (0.02 μg/L and 0.08 μg/L), DM did not cause obvious histopathological damage to hepatopancreas and gill tissue, while at high concentration (0.32 μg/L), the histopathological harm was serious and the damage did not recover to the initial level after 7-day decontamination. 0.02 μg/L DM exposure did not induce significant changes in most of the biomarkers except the increased lactate dehydrogenase (LDH) activity, lactic acid (LD) level, and the first increased then decreased mRNA expression of immune-related genes, indicating the stimulation of DM on energy production and immunity. 0.08 μg/L and 0.32 μg/L DM exposure resulted in varying degrees of damage on prawns, but overall, their toxic effects showed similar trends based on the biomarkers. Increase in malonaldehyde (MDA) and hydrogen peroxide (H 2 O 2 ) content and decrease in superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity after DM exposure demonstrated the oxidative stress caused by DM. The significantly increased acid phosphatase (ACP), alkaline phosphatase (AKP), LDH activity and LD level indicated hepatopancreatic dysfunction and respiration disruption. The first increased and then decreased expression pattern of immune-related genes indicated the immunosuppression caused by DM. After 7-day decontamination in freshwater, the activity/level of the biomarkers partly recovered. This study revealed the severe toxic effect of DM on Macrobrachium rosenbergii based on multiple biomarkers, providing fundamental knowledge for the establishment of DM toxicity assessment system with proper parameters in freshwater crustaceans., (Copyright © 2021. Published by Elsevier Inc.)

Sea anemones of the genus Bunodosoma possess along their body column, longitudinally arranged brown-colored vesicles. We have shown that in B. cangicum, these warty structures contain a mixture of potent toxins. This work highlights the neuro-inhibitory effects exhibited by two decapod crustacean species exposed to the extracts from these vesicles. For this, we use the unrefined toxin in doses, exposure times, and different exposure pathways. The findings show that at least one neuro-inhibitory compound is present and remains active regardless of the exposure method or dose tested. This toxin affects neuro-motor pathways but not neuro-sensory pathways. Shrimp exposed to toxin could continue to perceive and track food pellets but could not secure and consume their ration. Of six anatomical reflexes tested under the toxin's influence, voluntary movements of the mouthparts were impacted most commonly. Interestingly, all subject animals recovered from the toxin exposure within 2 h. Finally, we propose Reflexive Action Analysis (RAMP) as a tool to evaluate the potency of other neurotoxic or neuro-inhibitory compounds in crustacea. This work is the first to show the neuro-inhibitory activity of extracts from these sea anemone columnar vesicle structures and the first to evaluate these effects using RAMP reflex analysis., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Aquatic environmental pollutants have various impacts on aquaculture. Specifically, sulfide has been established as being toxic to aquatic animals including the oriental river prawn Macrobrachium nipponense. In response, the hepatopancreas has been broadly studied, as it plays a pivotal role in arthropod nutrient digestion and absorption, energy supply, and organ development as well as in crustacean immunity. However, the underlying molecular mechanisms of hepatopancreas's response to sulfide toxicity are still poorly understand. Herein, we used Nova-seq 6000 platform to conduct a comparative transcriptome analysis of gene expression profiles in the hepatopancreas of M. nipponense, while it was under the influence of a semi-lethal sulfide concentration (3.20 mg/L at 48 h). A total of 139 million raw reads were obtained, in which 67,602 transcripts were clustered into 37,041 unigenes for further analysis. After constant sulfide exposure for 48 h, 235 differentially expressed genes, i.e., DEGs (151 up-regulated and 84 down-regulated) were identified in the sulfide treatment group (TGHP) compared with the control group (CGHP). We used GO and KEGG databases to annotate all the DEGs into 1180 functions and 123 pathways, respectively. The metabolic pathways included proximal tubule bicarbonate reclamation, sulfur metabolism, glycolysis and gluconeogenesis, and the TCA cycle; while immune-related pathways contained Ras, Rap1, focal adhesion and platelet activation. Additionally, apoptosis-involved pathways e.g., lysosome, also exhibited remarkable alteration in the presence of sulfide stress. Notably, responses to external stimuli and detoxification genes- such as GSKIP, CRT2, APOD, TRET1, CYP4C3 and HR39- were significantly altered by the sulfide stress, indicating that significant toxicity was transferred through energy metabolism, growth, osmoregulatory processes and immunity. Finally, we demonstrated that in the present of sulfide stress, M. nipponense altered the expression of detoxification- and extracellular stimulation-related genes, and displayed positive resistance via tight junction activation and lysosome pathways. The results of these novel experiments shed light on the hepatopancreas's molecular response to sulfide stress resistance and the corresponding adaptation mechanism; and enable us to identify several potential biomarkers for further studies., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background and Objective: Effects of Cymbopogon citratus essential oil (EO) was tested on minimizing handling stress in Macrobrachium rosenbergii through the evaluation of their metabolite responses [glucose, lactate, glycogen, protein, Lactate Dehydrogenase (LDH), Malate Dehydrogenase (MDH), Acetylcholinesterase (AChE) and Alanine Aminotransferase (ALT)]. This study aimed to investigate the efficacy of C. citratus extract in the anaesthetization of M. rosenbergii., Materials and Methods: Three treatments including control, prawn exposed to stress alone (T1) and prawn exposed to stress in the presence of C. citratus EO (T2) were tested. A C. citratus EO at 500 μL L-1 had been determined in a previous study and was selected as the critical dose to be applied as an anesthetic agent. Handling stress was induced into prawns by netting, at 2 min interval for 30 min and their hemolymph were collected to determine the metabolite responses., Results: The increase of glucose, lactate and LDH of M. rosenbergii when exposed to handling stress alone (T1) in comparison to T2 (stress with anesthetic C. citratus EO) were identified. Further, a low glycogen level in parallel with low AChE activity was observed which indicates the involvement of secondary metabolites to cope with the energy demand in T1 over T2., Conclusion: This study indicates the efficiency of C. citratus EO to reduce stress during handling in M. rosenbergii.

The marine red algae Asparagopsis armata is an invasive species gaining competitive advantage by releasing large amounts of toxic compounds to the surrounding invaded area. The main objective of this study was to evaluate the effects of this invasive seaweed on marine invertebrates by exposing the common prawn Palaemon elegans and the marine snail Gibbula umbilicalis to the exudate of this seaweed. The seaweed was collected and placed in a tank for 12 h in the dark in a 1:10 ratio. Afterwards the seawater medium containing the released secondary metabolites was collected for further testing. Lethal and sublethal effects of A. armata were investigated. Biochemical biomarker responses associated with energy metabolism (lactate dehydrogenase, LDH; electron transport system activity, ETS; lipid, protein and carbohydrate content) were analysed. The biomarker responses showed physiological status impairment of invertebrates after exposure to low concentrations of this algal exudate. The highest concentrations of exudate significantly increased lipid content in both organisms. In the shrimp, protein content, ETS, and LDH were also significantly increased. By contrast, these parameters were significantly decreased in G. umbilicalis . A behavioural impairment was also observed in G. umbilicalis exposed to A. armata exudate, reducing feeding consumption. These results represent an important step in the research of natural toxic exudates released to the environment and prospective effects of this seaweed in invaded communities under increasing global change scenarios.

The hepatopancreas is the digestive organ of crustaceans, and plays important roles also in the synthesis and secretion of sexual hormones, immunological defenses and xenobiotic detoxification. Although the importance of this organ in crustaceans cannot be underestimated, the effects of ultraviolet B (UVB) radiation on hepatopancreas are poorly understood. Moreover, Macrobrachium prawns, have a transparent carapace, which make them more susceptible to UVB radiation, since their internal organs, such as hepatopancreas, are easily reached by solar radiation. Therefore, we aimed to evaluate UVB radiation toxicity on the morphology and morphometry of hepatopancreatic epithelial cells, and to investigate these UVB effects in subcellular compartments of the ecologically-important freshwater decapod, Macrobrachium olfersii. Hepatopancreas from the UVB-irradiated group showed a granular cytoplasm, with non-defined cell limits. Morphometric analyses revealed that the UVB-irradiated group exhibited a higher frequency of fibrillar (F-cell), resorptive (R-cell) and midget (M-cell), and decreased the blister-like (B-cell). It was also observed increased vacuole frequencies and increased F-, B- and R-cell volumes in the UVB-irradiated group. In addition, it was observed increased B-cell vacuolar volumes and decreased R-cell vacuolar volumes. Ultrastructural alterations occurred in subcellular compartments in F- and R-cells, e.g. loss of mitochondrial crests, morphologically compatible with mitochondrial fission, rough endoplasmic reticulum cisternae dilation, dilation of Golgi lamellar sacs, and increased vacuole and concentric membrane formation in the UVB-irradiated group. Our data showed that the hepatopancreas is an important target of UVB radiation, as demonstrated by a series of organ-specific morphological and morphometric impairments. Therefore, cell damage caused by UVB radiation can compromise metabolic functions in epithelial cells from the hepatopancreas, potentially affecting absorption, secretion and digestion processes, vitellogenin synthesis, immune responses and xenobiotic detoxification., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Many early stages of estuarine species congregate at the surface or in the upper mixing layer making them prone to UV light exposure and oil sheens. Laboratory testing was used to assess UV-oil sheen interactions with grass shrimp (Palaemon pugio). Newly hatched grass shrimp larvae were exposed to a 1-μm thick oil sheen for 24 h with or without an 8-h pulse of UV light. Grass shrimp were then transferred to clean seawater and non-UV conditions to measure development, growth, and reproductive fitness. Minimal toxicity was observed after the initial exposure but larval development was significantly delayed in shrimp exposed to the UV enhanced sheen. After reaching sexual maturity, shrimp were paired to evaluate effects on reproduction. Shrimp initially exposed to the UV enhanced sheen as larvae had a significant reduction in fecundity compared to controls. This demonstrates the importance of examining interactions between UV light and oil since negative effects to aquatic organisms may be underestimated if based on standard laboratory fluorescent lighting. Acute exposures of early life stages to thin oil sheens and UV light may lead to long-term impacts to individuals and ultimately to grass shrimp populations., (Published by Elsevier B.V.)

Contamination by sunscreens has become a serious environmental problem due to the increasing use of these products in coastal regions. Their complex chemical composition supposes an input of different chemical compounds capable of producing toxic effects and repelling organisms. The aim of the current study was to experimentally check the repellency of three commercial sunscreens [A (lotion), B (gel) and C (milk spray)] by assessing the escape (displacement towards areas with lower sunscreen levels) of the estuarine shrimp Palaemon varians exposed (4 h) to a gradient (0-300 mg/L) of the sunscreens in a heterogeneous non-forced exposure scenario. Additionally, mortality and immobility (72 h) were checked in a traditional forced exposure scenario. Considering that the toxicity of sunscreens is a little controversial regarding their chemical availability in the medium, two different methods of sunscreen solubilisation were tested: complete homogenization and direct immersion. Very low mortality was observed in the highest concentration of sunscreens A and C applied by direct immersion; however, for sunscreen B, the main effect was the loss of motility when homogenization was applied. Repellency was evidenced for two sunscreens (A and B) applied by direct immersion. The homogenization in the medium seemed to lower the degree of repellency of the sunscreens, probably linked to the higher viscosity in the medium, preventing the motility of shrimps. By integrating both short-term responses (avoidance and mortality/immobility), the PID (population immediate decline) calculated showed that avoidance might be the main factor responsible for the reduction of the population at the local scale., 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 Ltd. All rights reserved.)

The Giant freshwater prawn (Macrobrachium rosenbergii) breeds when in captive conditions. The eggs of a clutch are attached to the abdomen of berried females. Zinc oxide (ZnO) is one of the most important metal oxide-nanoparticles (NPs) that is widely used in various industries and is released into aquatic environments from wastewater management facilities. The present study was conducted to evaluate effects of ZnO on values for the reproductive variables: larvae development, crustacean hyperglycemic hormone (CHH) release from the X-organ into the hemolymph and anti-oxidative enzymes activity of M. rosenbergii. There were five groups including a group not treated (control), and groups treated with10, 20, 50, 100 mg/L ZnO in triplicate during a 90-day period. Results indicated that ZnO-NPs have marked effects on reproductive performance, offspring development, CHH release from the X-organ into the hemolymph and anti-oxidant enzymes activities with there being no spawning of brood-stock in the 100 mg/L ZnO group and in the prawns treated with 50 mg/L there was spawning but there was larvae mortality immediately subsequent to hatching. Also, values for viability rate of eggs, dry weight of eggs, brood-stock inter-spawn period and egg clutch somatic index (ESI) reproductive variables were affected by the NP. This NP was found to have a dose-dependent effect on CHH release from the X-organ into the hemolymph and also superoxide dismutase (SOD) and catalase activities in M. rosenbergii. The results indicate that M. rosenbergii, a freshwater decapod crustacean, is an appropriate species to study nano-material effects on reproduction in freshwater ecosystems., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Tonalide or acetyl hexamethyl tetralin (AHTN) is used as a fragrance additive in various household products. Recently, AHTN has drawn attention owing to its negative health effects on aquatic organisms. Data on AHTN toxicity toward aquatic species are limited. Therefore, this study tested the oxidative stress induced by AHTN exposure on the Rhodeinae sinensis Gunther and Macrobrachium nipponense. In this study, malonaldehyde (MDA) content and the activities of acetyl cholinesterase (AchE), superoxide dismutase (SOD), glutathione S-transferase (GST), and catalase (CAT) in R. sinensis Gunther were tested after 30 days of exposure to 30.093, 34.005, 38.426, 43.421, 49.067, 55.444, 62.652, 70.800, and 80.000 μg/L AHTN, respectively. The MDA, AchE, SOD, GST and CAT in M. nipponense were tested after 40 days of exposure to 60.000, 72.000, 86.400, 103.680, 124.416, 149.299, 179.159, 214.991, and 257.989 μg/L AHTN, respectively. In addition, an integrated biomarker response (IBR) index was utilised to evaluate the integrated toxic effects of AHTN on R. sinensis Gunther and M. nipponense. Finally, the predicted no-effect concentrations (PNECs) of AHTN, based on reproduction, biochemistry, survival, chronic toxicity, and acute toxicity endpoints were derived. The results indicated that low concentrations of AHTN can induce significant changes of oxidative stress biomarkers. The no observed effect concentrations (NOECs) of SOD, GST, AchE, CAT, and MDA were 103.680, 72.000, <60.000, 72.000, and <60.000 μg/L for R. sinensis Gunther and 38.426, 43.421, 30.093, 30.093, and 38.426 μg/L for M. nipponense, respectively. The IBR calculation results showed that 149.299 μg/L AHTN caused the highest toxic effect on R. sinensis Gunther after 30 days of exposure, whereas 70.797 μg/L AHTN caused the greatest damage to M. nipponense after 40 days of exposure. The PNECs of AHTN based on the non-traditional endpoints of biochemistry and reproduction were 0.00145 μg/L and 0.000395 μg/L, respectively, which were significantly lower than the PNEC of 2.636 μg/L for traditional endpoint survival. Therefore, the protection of aquatic organisms based on non-traditional toxicity endpoints should be considered in ecological risk assessment., 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 Inc. All rights reserved.)

The oriental river prawn, Macrobrachium nipponense, is a commercial freshwater prawn species in China. It is highly sensitive to hypoxia, and this has posed a challenge to its intensive culturing. To date, the effects of hypoxia on reproduction in female prawns are not entirely clear, as are the underlying mechanisms of the effects of hypoxia. In this work, comparative transcriptome and gene expression analyses of the eyestalk were performed in M. nipponense females under hypoxia and reoxygenation conditions. Sequencing and de novo assembly of the combined reads yielded 43,583 unigenes with an average length of 1726 bp. A total of 711 genes were found to be differentially expressed in the eyestalk under the hypoxia and reoxygenation conditions. With the help of functional and pathway enrichment analysis of the differentially expressed genes, a novel set of transcripts that were associated with several important functions, such as hormone biosynthesis and progesterone-mediated oocyte maturation, were identified. Additionally, ten neuropeptides were identified based on the differentially expressed transcripts, and they were validated by quantitative real-time polymerase chain reaction (qRT-PCR) and reverse transcription PCR (RT-PCR) analyses. Three neuropeptide genes were expressed in the neural tissue and ovary of the prawns; this indicates that they were involved in reproductive processes. In particular, RNA interference (RNAi) short neuropeptide F dramatically promoted ovary maturation, as indicated by the gonad somatic index. While the present findings do indicate that hypoxia affects reproductive function in M. nipponense females, in-depth functional analyses of the candidate neuropeptides should be conducted in the future to understand their role in hypoxia adaptation and the associated mechanisms that affect the reproductive capacity of this species., Competing Interests: Declaration of Competing Interest The authors state that this research is free of conflicts of interests., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Tyramine (TA), a biogenic monoamine, plays various important physiological roles including immunological regulation in invertebrates. In this study, the effects of TA on the regulation of immune resistance, carbohydrate metabolism and biogenic monoamine, as well as its signaling pathway in Macrobrachium rosenbergii were determined. Results showed that total haemocyte count, hyaline cells, semigranular cells, and phenoloxidase activity per 50 μL of haemolymph and per granulocyte (the sum of semigranular and granular cells) at 0.5 h as well as phagocytic activity and clearance efficiency to Lactococcus garvieae at 1 h of prawn injected with TA at 1 nmol prawn -1 significantly increased, but the significantly decreased plasma lysozyme activity, phagocytic activity, clearance efficiency, and haemolymph glucose and dopamine were observed in prawn injected with TA at 10 nmol prawn -1 for 0.5 h. Respiratory bursts and haemolymph lactate in two TA-injection treatments at 0.5 h and 0.5-1 h, respectively, were significantly higher than those of the saline control, and in addition, TA depressed dopamine release in a dose-dependent manner after 0.5 h of TA injection. All the examined parameters returned to control levels after prawn injected with TA for 2 h. The inhibited effect of TA (at 10 nmol prawn -1 injection) on the phagocytic activity and clearance efficiency to pathogens was blocked by prazosin (an α1 adrenoceptors antagonist). For prawn received TA for 1 h then challenged with Lactococcus garvieae at 2 × 10 5  colony-forming units prawn -1 , the survival ratio of TA 1 nmol prawn -1 -injected prawn significantly increased by 20%, compared to the saline-challenged control or TA 10 nmol prawn -1 -injected prawn after 144 h of challenge. These results suggested that the level of dopamine release suppression regulated by TA resulted in the immunoenhancing or immunosuppressive effects in prawn, and the signaling pathways of TA in mediating immune function were through octopamine (OA)/TA receptors., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Glyphosate-based herbicides (GBH), including Roundup®, are the most used herbicides in agricultural and non-agricultural areas, which can reach aquatic environments through drift during application or surface runoff. Some studies, mostly in fish, demonstrated that GBH caused oxidative stress in non-target animals. However, only few information is available on the GBH effects in the antioxidant and stress proteins of many other organisms, such as freshwater crustaceans. Thus, we aimed to investigate the effects of environmentally relevant GBH concentrations on the relative transcript expression (RTE) of the superoxide dismutase (sod1), catalase (cat), selenium-dependent glutathione peroxidase (gpx), glutathione-S-transferase (gst), thioredoxin (txn), heat shock protein (hsp70 and hsp90) in the hepatopancreas of the ecologically important freshwater prawn Macrobrachium potiuna. Moreover, this study aimed to assess the gender-differences responses to GBH exposure. Male and female prawns were exposed to three Roundup WG® concentrations (0.0065, 0.065 and 0.28 mg of glyphosate/L) and a control group (0.0 mg/L) for 7 and 14 days. In general, males had an under-expression of the studied genes, indicating an oxidative stress and possible accumulation of ROS in the hepatopancreas. In the opposite, females had an overexpression of the same genes, indicating a more robust antioxidant system, in order to cope with the possible ROS increase after Roundup WG® exposure. Therefore, results confirmed that gender could be a confounding factor in ecotoxicological assessment of GBH effects. Additionally, this work highlights that sod1, cat, gpx, gst, txn, hsp70 and hsp90 gene expressions seem to be useful biomarkers to investigate the oxidative stress caused by Roundup WG® in Macrobrachium sp., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The Pantanal (Brazil) is a wetland region characterized by seasonal flooding. Hydrological cycles influence the water physicochemical parameters, causing seasonal variations in pH and nitrites. Thus, the objective of this study was to evaluate the effects of varying pH and nitrite concentrations on the toxicity of the cypermethrin-based pesticide Barrage®, considering both lethal (mortality) and sublethal endpoints (growth and development). Larvae of the endemic shrimp Macrobrachium pantanalense and of the estuarine Amazonian congener Macrobrachium amazonicum were exposed to cypermethrin (through Barrage®) under several pH levels (6.5, 7.5 and 8.5) or nitrite concentrations (0.1, 0.2 and 0.4 mg/L). The pH had direct effects on all the tested endpoints for both species. For M. pantanalense, the lethal effects of the cypermethrin formulation were more pronounced at low pH (96-h LC 50  = 0.004 μg/L at pH 6.5, and 0.146 μg/L at pH 8.5). For M. amazonicum, an opposite response was observed, with increased toxicity of the formulation at high pH (96-h LC 50  = 0.110 μg/L at pH 6.5 and 0.044 μg/L at pH 8.5). Variations in pH also seemed to modify the sublethal effects of the formulation on larval growth and development of M. pantanalense. Nitrite concentrations affected larval growth of both species, modifying also the effects of the cypermethrin formulation on the larval development of M. amazonicum. This work shows the importance of considering abiotic factors for risk assessment either due to possible direct effects on the physiology of organisms and/or due to interactions with other stressors, particularly in fragile biomes such as Pantanal., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Ammonia nitrogen elevated is one of the commonest problem in the aquatic system, which caused a great threat to the survival and growth of prawn. However, little is know about the ammonia metabolism and detoxification strategy of prawn. In this study, the effects of ammonia-N (0, 0.108, 0.216, 0.324, or 0.54 mg L -1 ) on growth and metabolizing enzymes in hepatopancreas of Macrobrachium rosenbergii, including glutamine synthetase (GS), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and glutamate dehydrogenase (GDH), were investigated. The metabolome of its muscle was also analyzed after exposure to ammonia-N (0, 0.108, 0.324, or 0.54 mg L -1 ) for 20 days. The survival rate of M. rosenbergii decreased significantly after treatment with 0.54 mg L -1 ammonia-N compared with that in the other groups. However, ammonia-N had no significant effect on the growth of the river prawn after exposure for 20 days. GS activity increased significantly after exposure to 0.108 mg L -1 ammonia-N compared with the control and other ammonia-N-treated groups. Hepatopancreatic GDH activity was lower in the prawns treated with 0.216, 0.324, or 0.54 mg L -1 ammonia-N than in the control by 34.70%, 38.80%, or 41.94%, respectively. Ammonia-N had no significant effect on hepatopancreatic AST or ALT activity. Urea nitrogen was higher in the prawns treated with 0.216 mg L -1 ammonia-N than in the control or those treated with 0.54 mg L -1 ammonia-N. Ammonia-N had significant effects on the lipid, carbohydrate. and protein metabolism of M. rosenbergii, including purine metabolism, amino sugar and nucleotide sugar metabolism, α-linolenic acid metabolism, arginine and proline metabolism, glutathione metabolism, and phosphonate and phosphate metabolism, and on the terpenoid biosynthesis, lysine degradation, and lysine biosynthesis pathways. High concentrations of ammonia-N stress increased the content of glutamate and arginine, which may participate in the urea cycle, which synthesizes glutamine or urea to eliminate ammonia toxicity., (Copyright © 2019. Published by Elsevier Inc.)

Open-air landfill's may be are considered as a potential source of human environmental exposure to chemical substances such as, polycyclic aromatic hydrocarbons and toxic metals. Due to possible availability of mercury in the environment caused by open landfill emissions, this study evaluates the spatiality and seasonality of macroinvertebrates, in particular shrimps (Macrobrachium amazonicum), exposure to mercury (Hg). Information regarding Hg accumulation in this crustacean may be important for the development of public policies aiming conservation and preservation of ecosystems surrounding landfills in Amazon, and around the world. Sampling occurred quarterly in the following months: November/2015; February/2016; May/2016 and; August/2016. In each of these months, three points were selected: P1, P2 and P3. The samples were processed via acid digestion and the quantification of metal was performed by inductively coupled plasma mass spectrometry. The average concentration of total mercury (T-Hg) was 24.565 ± 6.610 µg kg -1 wet weight, with minimum and maximum limits of 12.742 ± 11.367 (P3) and 35.509 ± 14.761 µg kg -1 wet weight (P1) in November/2015 and August/2016, respectively. The concentration of total mercury (T-Hg) in shrimps was different between points (p = 0.004) and months (p = 0.000). The T-Hg concentrations were significantly higher in May and August 2016, which corresponds to the dry season. The presence of landfills promotes large accumulation of T-Hg in the aquatic biota and represents a risk to human health. However, seasonal changes in T-Hg levels were observed. In the wettest period, bioconcentration factor levels decrease in aquatic organisms.

Accumulation of nonylphenol (NP) in hepatopancreas, gonad, eyestalk, and muscle of freshwater prawn Macrobrachium rosenbergii following 72 h exposure to 100 µg/L NP, and depuration of NP in these tissues at 0.5-192 h post exposure were examined. We also examined the expressions of vitellogenin (Vg) and vitellogenin receptor (VgR) of prawn following 0-20 days exposure to 0, 1, 10, and 100 µg/L NP. NP accumulation in hepatopancreas and gonad with high concentration, and low concentration in muscle, but depurated faster in eyestalk and muscle. The expressions of vitellogenin (Vg) and vitellogenin receptor (VgR) increased directly with dose and time. In conclusion, NP accumulated significantly in gonad together with high Vg and VgR expressions, and depurated slow in hepatopancreas and gonad when prawns were removed back to control water. The induction of Vg and VgR under NP exposure might be a stress response in M. rosenbergii.

To test the toxic effects of tributyltin (TBT), Macrobrachium rosenbergii were exposed to three concentrations of TBT viz. 10 ng/L, 100 ng/L and 1000 ng/L for 90 days. The bioaccumulation of TBT level varied in hepatopancreas based upon dose dependent manner. Histopathological results revealed the reduction in basement membrane thickness, disruption of the hepatopancreatic tubules and abnormal lumen in hepatopancreas of TBT treated prawns. The ultrastructure of the control prawn showed normal architecture of cellular organelles with prominent nuclei in hepatocytes. On the other hand, many vacuoles, irregular arrangements of microvilli, swollen mitochondria, distorted rough endoplasmic reticulum cisternaes and abnormal nucleus were seen in the TBT treated group. Further, the biochemical and vitellogenin content were altered remarkably due to TBT exposure. It directly indicated that TBT had conspicuously inhibited the vitellogenesis. Therefore, it was inferred that the administration of TBT has considerably affected the hepatopancreatic functions in M. rosenbergii.

Recent research indicates that microplastic (<5 mm) ingestion may impact the immune function of marine and aquatic organisms at the tissue and cellular levels; however, their susceptibility to disease following exposure has not been directly investigated. The objective of the present study was to directly evaluate the impact of microplastic ingestion on the susceptibility of the grass shrimp Palaemonetes pugio to bacterial infection with Vibrio campbellii. Grass shrimp were exposed to one of several particle treatments (natural sediment, polyethylene spheres, polypropylene fragments, tire fragments, and polyester fibers) or particle-free water for 96 h at a nominal concentration of 50 000 particles/L prior to a bacterial challenge with V. campbellii. No significant mortality was observed among any of the particle types during the 96-h particle exposure. The survival of grass shrimp following V. campbellii challenge did not vary significantly among shrimp exposed to particle-free water, sediment, polyethylene spheres, polypropylene fragments, tire fragments, and polyester fibers. Grass shrimp cleared the majority of ingested particles and all the ventilated particles within 48 h. The present study shows that microplastic ingestion did not alter the susceptibility of grass shrimp to bacterial infection, and also provides depuration rates for a variety of microplastic shapes and polymer types that were previously lacking. This information increases our understanding of the size- and shape-dependent effects of microplastic ingestion. Environ Toxicol Chem 2019;38:2233-2242. © 2019 SETAC., (© 2019 SETAC.)

The oriental river prawn, Macrobrachium nipponense, is an economically and nutritionally important species of the Palaemonidae family of decapod crustaceans. Ammonia is a major aquatic environmental pollutant that negatively affects the health of prawns and their associated commercial productivity. Here, we used high-throughput sequencing techniques for detecting the effects of ammonia stress (22.1 mg/L ammonia-N for 48 h) on gene expression in the hepatopancreas of M. nipponense. We generated 176,228,782 high-quality reads after eliminating adapter sequences and filtering out low-quality reads, which were assembled into 63453 unigenes. Comparative analysis of the expression profiles of the ammonia-treated and control groups identified 887 differentially expressed genes (P < 0.05), including 481 upregulated genes and 406 downregulated genes. Analyses of the GO and KEGG databases revealed significant differences between the two groups in 32 pathways. Immune-related pathways under ammonia stress included Complement and coagulation cascades, Platelet activation, B cell receptor signaling pathway, Antigen processing and presentation, Chemokine signaling pathway, NOD-like receptor signaling pathway, RIG-I-like receptor signaling pathway, T cell receptor signaling pathway and Toll-like receptor signaling pathway. Remarkably, ammonia stress altered the expression patterns of key immune genes (lectin3, syntenin, alpha-2-macroglobulin, cathepsin L, PIM3, serine protease inhibitor, suppressor of cytokine signaling-2 like protein), indicating that ammonia-stress induce immune response. These data provide new insights into the immune response of M. nipponense and pave a new way for fighting ammonia stress. The genes and pathways identified here represent valuable genetic resources for development of molecular markers and genetic breeding studies., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

White spot disease caused by white spot syndrome virus (WSSV) is responsible for harming shrimp aquaculture industry and results in a pandemic throughout the world. Cathelicidin 5 treatment enhanced immune parameters including antioxidant enzyme activity and immune-related genes expression in shrimp Exopalaemon modestus. Shrimp treated with cathelicidin 5 and inoculated with white spot syndrome virus (WSSV) exhibited a significantly lower mortality rate and lower viral VP28 amplification and expression than control. This study addresses the role of cathelicidin 5 in immune stimulatory and antiviral activities that could protect E. modestus from WSSV infection., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

We examined the effects of exogenous dopamine on gill (Na + , K + )-ATPase activity in vitro in microsomal preparations from juvenile or adult freshwater shrimp, Macrobrachium amazonicum. Dopamine had no effect on enzyme activity in juveniles but stimulated activity in adult shrimp gills by ≈35%. Stimulation of the gill (Na + , K + )-ATPase in adult shrimps by 100 mmol L -1 dopamine was characterized kinetically by varying ATP, MgATP, and Na + and K + concentrations, together with inhibition by ouabain. Dopamine stimulated ATP hydrolysis by ≈40% obeying Michaelis-Menten kinetics, reaching V M  = 190.5 ± 15.7 nmol Pi min -1  mg -1 protein, K M remaining unaltered. Stimulation by Na + (≈50%) and K + (≈25%) revealed distinct kinetic profiles: although K M values were similar, Na + stimulation followed cooperative kinetics, contrasting with the Michaelian kinetics seen for K + . Stimulation by MgATP increased activity by ≈30% with little change in K M . Similar saturation profiles were seen for ouabain inhibition with very similar calculated K I values. Our findings suggest that dopamine may be involved in hemolymph sodium homeostasis by directly binding to the gill (Na + , K + )-ATPase at a site different from ouabain, thus stimulating enzyme activity in an ontogenetic stage-specific manner. However, dopamine binding does not affect enzyme affinity for cations and ouabain. This is the first report of the direct action of dopamine in stimulating the crustacean gill (Na + , K + )-ATPase., (Copyright © 2019. Published by Elsevier Inc.)

During the past decade, the total population and overall reproductive performance of the oriental river prawn (Macrobrachium nipponense) in the Anzali international wetland and its basin rivers has been severely reduced. This seems to be caused by an increase in heavy metal pollution from industrial wastewaters. We investigated the effects of chronic levels of lead (Pb) and cadmium (Cd) on reproductive success, hemato-immunology, crustacean hyperglycemic hormone (cHH), Pb/Cd bioaccumulation and histopathology of hepatopancreas of oriental river prawn. The present study was separately carried out using four chronic treatments for Pb (0 (control), 0.024, 0.072 and 0.12 μM/l) and Cd (0 (control), 0.026, 0.053 and 0.08 μM/l) in experimental tanks during 60 days. The broodstock were fed by commercial shrimp diet, ad libitum, twice a day. The results revealed that reproductive activities ceased when Pb levels were 0.072 and 0.12 μM/l and also when Cd levels reached 0.08 μM/l. At 0.024 μM/l Pb, 0.026 and 0.053 μM/l Cd, the prawns showed initial reproductive activity but after spawning, the egg-clutches were found detached from the pleopods of the brood females. Hemato-immunology results showed that by increasing the Pb and Cd concentrations, large granular hemocyte (LGH) and cHH values also increased. However, small granular hemocyte (SGH) count was inversely affected. A significant increase in mortality (P < 0.05) was observed as both Pb and Cd concentrations increased. Histological observations are indicative of increased necrosis, lumen size, melanization, vacuolation and abnormal lumen shape, as the heavy metals concentrations increased. In conclusion, obtained results can help to elucidate effects of Pb and Cd on freshwater decapod crustaceans reproductive performance and related physiological parameters., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Most aquatic animals use infochemicals from both conspecifics and heterospecifics to assess local predation risks and enhance predator detection. Released substances from injured conspecifics and other species (chemical alarm cues) are reliable cues to indicate an imminent danger in a specific habitat and often mediate the development of inducible defenses. Amphibian and fish embryos have been shown to acquire this information while at the embryonic stage of development, in relation to the developing nervous system and sensory development. With the exception of Daphnia , there is no information on chemically mediated responses to alarm cues in embryos of any crustacean groups. Therefore, we tested whether embryo exposure to chemical cues simulating predation on conspecifics or heterospecifics (closely related, non-coexisting species), or a mixture of both, alters embryonic developmental time, size and morphology of the first larval instar in Palaemon argentinus (Crustacea: Decapoda). Embryonic exposure to chemical alarm cues from conspecifics shortened the embryonic developmental time and elicited larger larvae with a longer rostrum. Rostrum length of the first larval instar changed independently of their size, thus elongated rostra can be considered a defensive feature. Embryonic developmental time was not altered by chemical alarm cues from either heterospecifics or the mixed cues treatment; however, exposure to these cues resulted in larger larvae compared with the control group. Chemically induced morphological plasticity in larvae in response to alarm cues from con- and heterospecifics suggests that such cues are conserved in palaemonids shrimps, providing embryos with an innate recognition of heterospecific alarm cues as predicted by the phylogenetic relatedness hypothesis., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Microcystins (MCs), produced by cyanobacteria, can strongly inhibit the activity of protein phosphatase, and exhibit strong hepatotoxicity. Macrobrachium rosenbergii is an important aquaculture economic species. Cyanobacterial blooms occur frequently during the culture of M. rosenbergii. However, the effects of MCs on the M. rosenbergii immune function have not been studied. In the present study, M. rosenbergii were exposed to environment-related concentrations of MC-LR type (0.5 and 5 μg/L) for 3 weeks. Hepatopancreatic histology was investigated, and antioxidant enzymes activity, acid phosphatase, alkaline phosphatase and lysozyme activity in hepatopancreas were also analyzed. Results showed that MC-LR could damage M. rosenbergii hepatopancreas, induce hepatopancreatic apoptosis and antioxidant dysfunctions. The expression profiles of major immune-related genes after MC-LR exposure were also detected. Some genes with antibacterial functions were suppressed, and the expression of apoptosis-related genes were up-regulated. After MC-LR exposure, the cumulative mortality of M. rosenbergii infected with Vibrio vulnificus and Aeromonas hydrophila were much higher than the control in a time-dose dependent manner. These results indicated the potential negative influence of MC-LR on the immune function of M. rosenbergii., (Copyright © 2019. Published by Elsevier Ltd.)

In order to study the effects of Moringa oleifera leaf extract on Macrobrachium rosenbergii under high ammonia exposure, freshwater prawns were randomly divided into five groups: a control group was fed with basal diet, and four treatment groups fed with basal diet supplemented with 0.25%, 0.5% and 1.0% M. oleifera leaf extract and 0.025% Enrofloxacin for 60 days, respectively. Then, freshwater prawns were exposed to high ammonia stress for 72 h and Vibro anguillarum infection. The growth, antioxidant capabilities, related immune genes as well as resistance to infection by V. anguillarum were determined. The results showed that compared with the control group, the weight gain, specific growth rate and protein efficiency rate, haemolymph catalase (CAT), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) increased while feed conversion ratio, haemolymph aspartate aminotransferase, alanine aminotransferase, nitrogen oxide (NO), hepatopancreas heat shock proteins (HSP70), immune deficiency (IMD) expression levels decreased in the group of 0.5% M. oleifera leaf extract before the stress. After ammonia stress, the group of 0.5% M. oleifera leaf extract also could improve the haemolymph SOD, glutathione peroxidase, NO, iNOS, hepatopancreas HSP70 expression levels and reduce haemolymph CAT, hepatopancreas peroxiredoxin 5 and NF kappa B inhibitor alpha expression level compared with the control group. The rate of mortality of the prawns challenged with V. anguillarum was lower in the supplemented groups in comparison with the control group with the lowest being in the group of 0.5% M. oleifera leaf extract. Antioxidant activities as well as biochemical parameters in the enrofloxacin group (0.025%E) were not significantly enhanced both pre and post challenge in comparison with the M. oleifera leaf extract groups, showing the superiority of the natural herb over the synthetic antibiotic. In summary, this study suggested that at an inclusion rate of 0.5%, M. oleifera leaf extract could increase the growth performance, even has positive effects on physiological and immune function and prevents high ammonia stress in the Freshwater prawn, M.rosenbergii., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Microcystins (MCs) are produced by cyanobacterial blooms and known for their hepatotoxicity. They could cause serious damage to the reproduction of higher vertebrate mice and fish. However, few studies have focused on the reproductive toxicity of MCs to invertebrates. Giant freshwater prawn Macrobrachium rosenbergii are highly cultivated in China. During their breeding process, M. rosenbergii are often infested by cyanobacteria blooms. In the present study, to investigate the toxic effect of MCs on the testicular development of M. rosenbergii. Male M. rosenbergii were exposed to environmental relevant concentration of MC-LR for 1, 2 and 3 weeks. Results showed that MC-LR entered M. rosenbergii testis, down-regulated hemolymph testosterone (T) levels, and damaged testicular germ cells, mitochondria and cell junctions, and inhibited testicular development. Moreover, MC-LR could significantly induce the expression of gonadal development related genes in testis and eyestalk). The present results indicate that MC-LR can disrupt the testicular development of M. rosenbergii by affecting T levels and gonadal development related genes in the testis and eyestalk., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Glyphosate-based herbicides (GBH), including Roundup, are the most widely used pesticides in the world. Glyphosate residues have been detected in surface and groundwater, in food, and in human blood and urine. The effects of this herbicide on different levels of biological organization are an important concern that needs to be investigated. In general, the toxicity of GBH in invertebrates is poorly understood, and it is the motivation of this study. Thus, the aim of this study was to evaluate cellular responses of the hepatopancreas, an organ involved in the detoxification process in invertebrates, after exposure to environmentally relevant concentrations of GBH, using prawn Macrobrachium potiuna as a model. Prawns were exposed to three concentrations of GBH (0.0065, 0.065 and 0.28 mg L -1 ) for 7 or 14 days. Alterations in the morphology of the hepatopancreas and in subcellular components of R cells, which are responsible for the detoxification process, were analyzed, and an index for subcellular alterations was standardized. GBH exposure induced tissue commitments on the hepatopancreas, as well as important impairments of R cells that could compromise the normal functioning of the cells, especially in the detoxification processes. The major cellular impairments were intense vacuolization, dilatation of the cisterns of the rough endoplasmic reticulum and Golgi bodies, increase of perinuclear space, necrosis, concentric membrane formation and mitochondria crest loss. Our data contribute to the knowledge of the cytotoxic effects of low GBH concentrations on aquatic invertebrates, specifically their effects on the hepatopancreas, an important organ for the metabolism of crustaceans. These results also indicate that concentrations considered safe by regulatory agencies should be reviewed to minimize the effects on non-target organisms. This study also contributes to the standardization of an ultrastructure index for the assessment of GBH in palaemonids, which could be used for the assessment of contaminants in crustaceans and other species with hepatopancreas., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A key question to be asked when developing regional water quality criteria with scarce toxicity data is whether such data need to be locally derived. To address this, ammonia toxicity data from local aquatic species in the Liao River were compared against data from species native and non-native to China, based on comparisons of the overall trends of species sensitivity distributions and derived water quality criteria. Liao River data were acquired by acute and chronic tests using five local freshwater invertebrate species, and then compiled alongside published data from Chinese national guidelines and international literature. Models of best fit using three species sensitivity distribution approaches (log-logistic, log-normal, and Burr III) did not vary markedly (r 2 >0.9), and no specific model provided a best fit across all data sets. The comparisons of the overall trend of species sensitivity distribution curves showed no significant differences at either a national (Chinese native taxa tested in China versus non-native taxa) or regional level (Liao River taxa versus non-Liao River taxa). The comparisons also revealed that the inclusion or exclusions of different ecological groups had little influence on the overall trends of species sensitivity distributions. These findings suggested data on non-local and non-native species, and data from local species tested elsewhere, could be appropriate for guiding the derivation of ammonia water quality criteria for regions such as Liao River. However, caution is needed when using hazardous concentration 5% values in the development of site-specific water quality criteria for a river basin due to the considerable variation observed for ammonia (16.8-56.6 mg/L), although these differences were not statistically significant. Based on the toxicity test evaluation, a preliminary acute value of 10.0 mg/L and chronic value of 1.7 mg/L (at pH of 7.0 and 20 °C) are proposed as site-specific ammonia water quality criteria for the Liao River, China., (Copyright © 2018. Published by Elsevier Inc.)

The oriental river prawn Macrobrachium nipponense is a highly adaptable, tolerant, and fecund freshwater prawn that inhabits a wide range of aquatic environments. The hepatopancreas of crustaceans is not only a site for secretion of digestive enzymes, and also plays important roles in several metabolic processes, such as lipid and carbohydrate metabolism. It is the main organ for the detoxification and immunity. In this study, high-throughput sequencing techniques were used to detect the effect of nitrite stress (10 mg/L nitrite-N for 48 h) on gene expression in the hepatopancreas of M. nipponense. A total of 13,769 million reads were harvested, and 94,534 transcripts were de novo assembled using Trinity software and produced 56,054 non-redundant transcripts. A total of 825 differentially expressed genes were obtained comparing 48 h nitrite stress with control group. In the analysis of GO and KEGG database, significant differences were found in 49 pathways. Immune-related pathways under nitrite stress included arginine and proline metabolism, glutamate metabolism, Jak-Stat signaling pathway, endocytosis, wnt signaling pathway, RIG-I-like receptor signaling pathway, TGF-beta signaling pathway, GnRH signaling pathway and phagosome. Apoptosis-related pathway was also significantly altered, such as lysosome and apoptosis. Remarkably, nitrite stress altered the expression patterns of key apoptosis genes (tetraspanins-like protein, LAMP, CD63, caspase 3C and Caspase 1) and immune genes (Serine proteinase-like protein, C-type lectin, daf-36, SOCS-2, alpha-2-macroglobulin), confirmed that nitrite-stress induce immune response and eventually even apoptosis. This study provided a new insight into the role of hepatopancreas in crustaceans, and further investigation will continue., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Pantanal shrimp Macrobrachium pantanalense was exposed for 96 h to the cypermethrin-based formulation Barrage ® . Population-relevant endpoints (survival, swimming behavior) as well as histopathology of gills were analyzed. A 96 h-LC 50 of 0.93 μg/L of cypermethrin was calculated while equilibrium disturbances were observed at 1.25 μg/L. Histological examination showed predominantly regressive changes in the gills of shrimp exposed to concentrations of 0.25 and 1.25 μg/L. Three levels of lesions were observed in the gills: I- Intercellular edema, epithelial lifting of the lamellae and lamellar fusion, fat vacuoles and hypertrophy of gill epithelial cells or mucous cells; II- nuclear changes, atrophy (reduction of volume and number) and hyperplasia of gill epithelia and III- necrosis. This study shows the high sensitivity of the shrimp M. pantanalense to the pesticide Barrage ® highlighting the importance of monitoring agrochemicals residues in the Pantanal region (Brazil) and conduct risk evaluation studies to prevent deleterious effects on the aquatic communities of Pantanal., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Giant freshwater prawn Macrobrachium rosenbergii is an economically important species. However, its growth retardant have brought serious economic losses in recent years. Antibiotics abuse is suggested as a reason for M. rosenbergii's growth retardant, while few studies focused on the toxic effect of antibiotics on M. rosenbergii. To investigate the effect of enrofloxacin, a widely used antibiotic, on juvenile M. rosenbergii, a 14 days exposure study was carried out within 0.2, 1 and 5 mg/L enrofloxacin and followed by 7 days decontamination. Results showed that during the test period, enrofloxacin had the largest accumulation in juvenile shrimp at day 3, and gradually decreased at day 7 and 14, and almost all the drugs are cleared after 3 days decontamination. Short-term exposure to low dose enrofloxacin can promote the growth of juveniles. High dose enrofloxacin inhibited the growth of juvenile shrimp, to gill and liver damage, and induced apoptosis of the hepatopancreatic cells. These adverse effects was possibly caused by enrofloxacin-induced oxidative stress. Moreover, we also found the damage caused by high concentrations of enrofloxacin was irreversible in the short term. Collectively, these data indicated that enrofloxacin did affect the juvenile shrimp growth and development, and high level enrofloxacin abuse may contributed to M. rosenbergii's growth retardant., (Copyright © 2018. Published by Elsevier Inc.)

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.

In the present study, an acute toxic test was performed to assess the oxidative stress and genotoxic effects of the herbicide on the freshwater shrimp Macrobrachium nipponensis. The results showed that the 48-h and 96-h LC 50 values of Roundup to M. nipponensis were 57.684 mg/L and 11.237 mg/L, respectively. For further investigation, the shrimps were exposed to sublethal concentrations of 0.35, 0.70, 1.40, 2.80 and 5.60 mg/L for 96 h. A significant decrease in total haemocytes count (THC) was observed at concentration of 5.60 mg/L throughout the experiment. The level of superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) in all the treatments decreased in a dose- and time-dependent manner except for the concentration group of 0.35 mg/L. The malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ) and protein carbonyl in serum increased significantly at concentrations of 2.80 mg/L and 5.60 mg/L. A significant decrease in acetylcholinesterase (AChE) activity was observed at each concentration (P＜0.05). In addition, the micronucleus (MN) frequency of haemocytes significantly increased (P＜0.05) at concentrations of 1.40, 2.80 and 5.60 mg/L, whereas the comet ratio and %DNA in the tails exhibited a clear time- and dose-dependent response during the exposure. The analysis of the integrated biomarker response (IBR) showed the induction of oxidative stress biomarkers and the inhibition of antioxidants, and this dose-dependent relation suggests the sensitivity and availability of all the biomarkers. These results revealed that Roundup had a prominent toxic effect on M. nipponensis based on the antioxidative response inhibition and genotoxicity., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Since the end of the 1980s, white shrimps (Palaemon longirostris) from the Gironde estuary have exhibited exoskeletal malformations, mainly involving cephalothorax, rostrum, scaphocerites and uropods. An 8-month study was carried out in 2015. Each month, 200 individuals were sampled and examined for exoskeletal malformations. Temporal variations in malformation frequency were noted, particularly during the breeding period, along with decreases in the size of non-deformed shrimps related to the appearance of juveniles in breeding sites, and high mortality among deformed shrimps. A significant increase in proportions of deformed shrimp was observed, relating particularly to the size (and therefore the age) of individuals. No significant difference was found between shrimp proportions with different numbers of malformations (one to four) for a fixed size class, nor was there any variation in proportions within different size classes for a fixed number of malformations. This would appear to indicate that the number of malformations is acquired and new malformations do not seem to appear during the life cycle, except for the smallest (youngest) shrimps. The malformation spectrum showed no significant differences between the biggest and smallest individuals for the different malformation associations, except for those involving cephalothorax, rostrum and uropods. This would suggest that some malformation associations lead to a higher mortality rate in shrimps subjected to them, due to greater impairment of feeding and/or swimming behaviour. Multiple component analysis of the different types of malformation showed correlations between exoskeletal pieces (rostrum and cephalothorax) and appendixes (scaphocerites and uropods). Regarding metal contamination in shrimp, no significant difference was highlighted between deformed and non-deformed shrimps. Organic pollutants were not measured in tissues. Certain herbicides such as metolachlore and chlortoluron were detected at high concentrations in the Gironde estuary during the breeding period corresponding to the higher occurrence of exoskeletal malformations.

Essential oil derivatives are widely used for anaesthetising aquatic animals. However, the effectiveness of anaesthesia often varies according to the anaesthetic agent, species, temperature, dosage, and interactions among these factors. This study evaluated the effects of eugenol on three sizes of the shrimp Palaemonetes sinensis at different concentrations and temperatures. Eugenol dose, water temperature, and shrimp size were found to significantly influence anaesthesia in P. sinensis. Induction time decreased linearly with increasing water temperature and eugenol concentration, while it increased with body weight. However, recovery times lengthened with increasing concentration and temperature, and shortened with lower body size. At 100 and 200 μL/L eugenol concentrations, the survival rates of medium and large shrimps were maintained at over 80% at all temperatures studied over 72 h recovery. However, the survival rates of small shrimps were below 60% at 24 °C and 28 °C over 5 days of recovery. These results suggest that eugenol is an effective and rapid anaesthetic for P. sinensis, but it might have disadvantages such as slow recovery and possible mortality in small shrimps and at higher temperatures and dosages.

The use of freshwater invertebrates as bioindicator of heavy metal pollution is an important tool for environmental biomonitoring. This study investigated antioxidant activities and DNA profile in two limicolous earthworms (Alma millsoni and Libyodrilus violaceus) and freshwater prawns (Macrobrachium vollenhovenii) at selected points of Ogun River, Abeokuta. Heavy metal concentrations and DNA profile in the earthworms and prawn tissues were measured using standard procedures. Zn concentration was higher than other heavy metals in A. millsoni (685.83±114.42mg/kg), L. violaceus (1913.3±1098.7mg/kg) and M. vollenhovenii (134.7±13.61mg/kg). Superoxide dismutase activity ranged from 62.44±7.16-79.82±11.18U/g tissues, 60.26±11.18-71.07±7.54U/g tissues and 74.07±16.69-87.79±8.50U/g tissues in A. millsoni, L. violaceus and M. vollenhovenii respectively. RAPD-PCR revealed varying DNA profile among the earthworms samples; the UPGMA dendrogram formed two distinct clusters at genetic similarity coefficient of 0.15-0.2 with one cluster consisting of Alma millsoni and Libyodrilus violaceus from Sokori, Enugada and Iberekodo sampling points and the second cluster forming two distinct sub-clusters comprising Arakanga and Ago-ika's L. violaceus in one and A. millsoni in the other. High genetic variability was recorded among the earthworm species while the freshwater prawn showed no variability. Antioxidant activities and genetic variability in earthworms could serve as biomarkers of heavy metal pollution in freshwater environment., (Copyright © 2017 Elsevier B.V. All rights reserved.)