The effect of banana blossom on growth performance, immune response, disease resistance, and anti-hypothermal stress of Macrobrachium rosenbergii.

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.

Comparative transcriptome analysis of the gills and hepatopancreas from Macrobrachium rosenbergii exposed to the heavy metal Cadmium (Cd2+).

Heavy metal Cadmium (Cd2+) 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 Cd2+ 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 Cd2+ 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 Cd2+ 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 Cd2+ 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 Cd2+ 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 Cd2+. This comparative transcriptome provides valuable molecular information on the mechanisms of responding to Cd2+ stress in M. rosenbergii, which lays the foundation for further understanding of heavy metal stress.

Assessment of deltamethrin toxicity in Macrobrachium nipponense based on histopathology, oxidative stress and immunity damage.

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 LC50 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 (H2O2), 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.

Effect of Cymbopogon citratus Essential Oil (EO) on Handling Stress in Giant Freshwater Prawn (Macrobrachium rosenbergii).

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.

Multi-biomarker assessment in the giant freshwater prawn Macrobrachium rosenbergii after deltamethrin exposure.

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 LC50 values to be 1.919, 0.603, 0.539, and 0.449 µg/L, respectively. According to 96 h LC50, 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 (H2O2) 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.

Comparative transcriptome reveals the response of oriental river prawn (Macrobrachium nipponense) to sulfide toxicity at molecular level.

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.

The effects of vesicle toxin from the sea anemone Bunodosoma cangicum on the behavior of a freshwater shrimp, Palaemon argentinus, and shore crab, Neohelice granulata.

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.

Effect of Fluoxetine Hydrochloride on Routine Metabolism of Lambari (Deuterodon iguape, Eigenmann, 1907) and Phantom Shrimp (Palaemon pandaliformis, Stimpson, 1871)

HIGHLIGHTS Fluoxetine increases the metabolic rate and excretion of ammonia in both species. O:N ratio in fish showed higher values in the highest concentrations of fluoxetine. The LC50 - 96 hour values of Palaemon pandaliformis represented greater toxicity. Both species are a good biological model for fluoxetine exposure studies.

Abstract Fluoxetine is an emerging pollutant that acts as a selective serotonin reuptake inhibitor (SSRI) and being a hydrolytic molecule that is photolytically stable and accumulaties in biological tissues, its disposal in the aquatic environment can interfere with the physiology of fish and shrimp. Thus, the objective of this study was to analyze the effects of fluoxetine on routine metabolism (metabolic rate, specific ammonia excretion and O:N ratio) of Deuterodon iguape and Palaemon pandaliformis. For this, five groups of each species, were exposed to different concentrations of fluoxetine for 24 hours (D. iguape) and 2 hours (P. pandaliformis). The results demonstrated that in D. iguape exposure to fluoxetine significantly increased both the metabolic rate by 75%, 85%, 55% and 50% for concentrations of 0.05; 0.1; 0.5 and 1.0 mgL-1, respectively, and the specific ammonia excretion by 40%, 48% and 20% for concentrations of 0.05; 0.1 and 0.5 mgL-1, respectively, when compared with their control. The O:N ratio was statistically greater in concentrations of 0.5 and 1.0 mgL-1. Concerning P. pandaliformis, exposure to fluoxetine increased metabolic rate at concentrations 30.0 and 60.0 µgL-1, and also increased specific ammonia excretion at concentrations 10.0, 30.0 and 60.0 µgL-1, when compared with the control group. It was concluded that exposure to fluoxetine increases the routine metabolism of both species and that at the concentration 1.0 mgL-1, Deuterodon iguape required different energy substrates.

Impacts of the Invasive Seaweed Asparagopsis armata Exudate on Energetic Metabolism of Rock Pool Invertebrates.

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.

Developmental and reproductive effects in grass shrimp (Palaemon pugio) following acute larval exposure to a thin oil sheen and ultraviolet light.

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.

Effects of ZnO nanoparticles on the Giant freshwater prawn (Macrobrachium rosenbergii, de Man, 1879): Reproductive performance, larvae development, CHH concentrations and anti-oxidative enzymes activity.

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.

Ultraviolet B radiation affects epithelial cell morphology and ultrastructure in the hepatopancreas of the freshwater decapod Macrobrachium olfersii.

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.

Repellency and mortality effects of sunscreens on the shrimp Palaemon varians: Toxicity dependent on exposure method.

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.

Antioxidative enzyme activities in the Rhodeinae sinensis Gunther and Macrobrachium nipponense and multi-endpoint assessment under tonalide exposure.

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.

The temporary modulation of tyramine on immune responses, carbohydrate metabolism, and catecholamines in Macrobrachium rosenbergii.

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 × 105 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.

Macrobrachium amazonicum (Crustacea, Decapoda) Used to Biomonitor Mercury Contamination in Rivers.

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.

Identification of neuropeptides from eyestalk transcriptome profiling analysis of female oriental river prawn (Macrobrachium nipponense) under hypoxia and reoxygenation conditions.

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.

Modulation of antioxidant gene expressions by Roundup® exposure in the decapod Macrobrachium potiuna.

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.

Effects of ammonia-N exposure on the growth, metabolizing enzymes, and metabolome of Macrobrachium rosenbergii.

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.

Effects of pH and nitrites on the toxicity of a cypermetrin-based pesticide to shrimps.

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 LC50 = 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 LC50 = 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.