The tale of an endemic shrimp's exceptional osmoregulation and the ancient Athalassic mangrove oasis.

The hyperarid mangrove in the Middle East is characterised by the absence of rivers or freshwater inputs and is one of the most extreme settings of this ecosystem on Earth. Endemic to Qatar's hyperarid mangroves, a Palaemon shrimp is uniquely confined to a sole mangrove site in the Arabian Gulf. Within these mangrove channels, we unveiled brine groundwater sources exceeding 70 ppt salinity, contrasting the local marine standard of 42 ppt. Concurrently, a mysid species typically linked to salt pans and groundwater coexists. Stable isotopic analysis implied the existence of a predator-prey dynamic between this mysid species and the studied shrimp. Then, investigating the endemic shrimp's adaptation to extreme salinity, we conducted osmolarity experiments and phylogenetic studies. Our findings demonstrate that this shrimp transitions from hypo- to hyper-osmoregulation, tolerating salinities from 18 to 68 ppt-an unprecedented osmoregulatory capacity among caridean shrimps. This speciation pattern likely arises from the species osmolarity adaptation, as suggested for other Palaemon congeners. Phylogenetic analysis of the studied Palaemon, along with the mangrove's geological history, suggests a profound evolutionary interplay between the ecosystem and the shrimp since the Eocene. This study proposes the hyperarid mangrove enclave as an Athalassic mangrove oasis-a distinctive, isolated ecosystem within the desert landscape.

Regulation of early spermatogenesis in the giant prawn Macrobrachium rosenbergii by a GCL homolog†.

The germ cell-less gene is crucial for gonad development in various organisms. Early interventions in its expression suggested a regulatory role at the mitotic stages of spermatogenesis, and its early knockout resulted in complete sterility in Drosophila. Genomic and transcriptomic data available for the catadromous giant prawn Macrobrachium rosenbergii enabled the identification of a germ cell-less homolog for this species, which we termed MroGCL (mRNA accession number OQ533056). An open reading frame containing 494 amino acids and a typical evolutionarily conserved BTB/POZ domain suggests possible protein-protein interaction functions in keeping with the Drosophila germ cell-less protein. Genomic mapping of MroGCL showed a full length of 120 896 bases. Analysis of the temporal expression of MroGCL showed constant expression in early prawn embryonic and larval stages, but a significant increase 10 days after metamorphosis when crucial sexual differentiation processes occur in prawns. In adult animals, high expression was detected in the gonads compared to the somatic tissues. RNAi-based knock-down experiments showed that both the silenced and control groups reached advanced spermatogenic stages, but that there was a significant decrease in the yield of spermatozoa in about half of the silenced animals. This finding supports our hypothesis that MroGCL is crucial for mitosis during early stage spermatogenesis. In conclusion, this study contributes to the understanding of crustacean gonad development and provides a stepping stone in the development of environmentally valuable sterile crustacean populations.

Metabolomic assessment of African snail (Achatina fulica) meal on growth performance of giant river prawn (Macrobrachium rosenbergii).

This study aimed to investigate the effects of replacing fishmeal (FM) with African giant snail (Achatina fulica) meal (SM) on the growth performance of giant river prawn (Macrobrachium rosenbergii), as well as to analyze the associated metabolomic changes. Six diets were formulated, replacing FM with SM at different inclusion levels ranging from 0 % to 100 %. Growth performance and feed conversion ratio of prawns fed diets with FM replaced by SM up to 80 % were not significantly different from control. In contrast, significantly decreased growth performance and higher feed conversion ratio (FCR) occurred with diets containing 100 % SM. To gain insights into the metabolic regulation of prawns fed different diets, a 1H NMR metabolomics approach was used to assess the metabolic changes in prawns fed diets containing 0 % and 80 % SM. The results revealed up-regulated metabolites significantly involved in several metabolic pathways, including alanine, aspartate, and glutamate metabolism; citrate cycle (TCA cycle); aminoacyl-tRNA biosynthesis; and valine, leucine, and isoleucine biosynthesis. These findings imply that including SM in the diet might modulate the regulation of muscle amino acids and tRNA synthesis, suggesting a potential impact on protein biosynthesis mechanisms. Additionally, alterations in the TCA cycle may reflect changes in carbon utilization, potentially contributing to the growth performance of giant river prawns when fishmeal is replaced with SM without adversely affecting their growth. In conclusion, this study demonstrated that SM could be a promising alternative protein source in aquafeed. The metabolomic approach provides valuable insights into the metabolic changes in prawns fed different diets, aiding in the development of more effective aquafeeds in the future. The study's limitations, such as the simplified diet formulation and the limited scope of the metabolomic analysis, were acknowledged and discussed, highlighting the need for further research to build upon these findings.

Pyrethroid insecticides pose greater risk than organophosphate insecticides to biocontrol agents for human schistosomiasis.

Use of agrochemicals, including insecticides, is vital to food production and predicted to increase 2-5 fold by 2050. Previous studies have shown a positive association between agriculture and the human infectious disease schistosomiasis, which is problematic as this parasitic disease infects approximately 250 million people worldwide. Certain insecticides might runoff fields and be highly toxic to invertebrates, such as prawns in the genus Macrobrachium, that are biocontrol agents for snails that transmit the parasites causing schistosomiasis. We used a laboratory dose-response experiment and an observational field study to determine the relative toxicities of three pyrethroid (esfenvalerate, λ-cyhalothrin, and permethrin) and three organophosphate (chlorpyrifos, malathion, and terbufos) insecticides to Macrobrachium prawns. In the lab, pyrethroids were consistently several orders of magnitude more toxic than organophosphate insecticides, and more likely to runoff fields at lethal levels according to modeling data. At 31 water contact sites in the lower basin of the Senegal River where schistosomiasis is endemic, we found that Macrobrachium prawn survival was associated with pyrethroid but not organophosphate application rates to nearby crop fields after controlling for abiotic and prawn-level factors. Our laboratory and field results suggest that widely used pyrethroid insecticides can have strong non-target effects on Macrobrachium prawns that are biocontrol agents where 400 million people are at risk of human schistosomiasis. Understanding the ecotoxicology of high-risk insecticides may help improve human health in schistosomiasis-endemic regions undergoing agricultural expansion.

Are there any effects of eco-climatic factors on freshwater prawn reproduction: a study on monsoon river prawn, Macrobrachium malcolmsonii.

Prawns are regarded as delicious food items and a source of income as it is the salient exportable fishery item of Bangladesh and a number of other countries. Climate change may inhibit the general physiological activity of prawns. This study explored the eco-climatic effects on reproduction, including size at first sexual maturity (Lm), spawning season, and fecundity of monsoon river prawn, Macrobrachium malcolmsonii (Milne-Edwards, 1844). The study was carried out in the Ganges (Padma) River basin, northwestern Bangladesh, from January to December 2014. Seven hundred fifty-seven specimens ranging from 0.80 to 18.27 g BW (body weight) with TLs (total length) of 4.20 to 11.52 cm were analyzed. The Lm was estimated as 6.2-6.6 cm. Total fecundity was higher than earlier studies and ranged from 2743 to 122,165 with a mean value of 6715 ± 2437. The spawning season was estimated from April to October with a peak in July. Significant correlation was found between rainfall and the presence of ovigerous females, i.e., spawning season. The number of ovigerous females increased in ≥ 100 mm average rainfall, and the maximum ovigerous female spawned at 250-320 mm rainfall. Spawning season of M. malcolmsonii may have shifted with shifting rainy season as a result of climate change. The findings of our investigation would be useful for sustainable management of the fishery of M. malcolmsonii and other prawn species in the Ganges River, Bangladesh, and adjoining countries with similar eco-climatic factors.

Sensitivity of cholinesterase activity in juvenile giant freshwater prawn (Macrobrachium rosenbergii de Man, 1879) to organophosphate diazinon.

Pesticides containing diazinon are frequently used in agriculture in the Vietnamese Mekong delta region leading to their potential residual occurrence in the environment. Under laboratory conditions, exposure to diazinon has been shown to result in adverse inhibition of cholinesterase enzyme (ChE) activity and subsequent death in several fish species. This study investigated a 96-h median lethal concentration (LC50) of diazinon concentrations from 0.08 to 1.25 mg/L on juvenile giant freshwater prawns (Macrobrachium rosenbergii de Man, 1879) in tanks in the laboratory. Inhibition of ChE in the flesh and in the eyes of the tested shrimps after exposed to diazinon concentrations of 2.7, 27, 67.5 µg/L equivalent to 1%, 10%, 25% LC50-96 h was calculated. The results indicated that diazinon was highly toxic to giant freshwater prawn with a low LC50-96 h of 270 µg/L. The activity of ChE in the flesh was more sensitive to diazinon than that in the eyes. Furthermore, in the future, the activity of ChE in the flesh or in the eyes of shrimps has potential to be used as biomarker for rapid recognition of diazinon contamination in water.

Sex-specific thermal tolerance limits in the ditch shrimp Palaemon varians: Eco-evolutionary implications under a warming ocean.

As global temperatures continue to rise due to global change, marine heatwaves are also becoming more frequent and intense, impacting marine biodiversity patterns worldwide. Organisms inhabiting shallow water environments, such as the commercially relevant ditch shrimp Palaemon varians, are expected to be the most affected by rising temperatures. Thus, addressing species' thermal ecology and climate extinction-risk is crucial to foster climate-smart conservation strategies for shallow water ecosystems. Here, we estimated sex-specific upper thermal tolerance limits for P. varians via the Critical Thermal Maximum method (CTmax), using loss of equilibrium as endpoint. We further calculated thermal safety margins for males and females and tested for correlations between upper thermal limits and shrimps' body size. To determine sex-biased variation in P. varians' traits (CTmax, weight and length), we compared trait variation between females and males through the coefficient of variation ratio (lnCVR). Females displayed an average CTmax value 1.8% lower than males (CTmaxfemales = 37.0 °C vs CTmaxmales = 37.7 °C). This finding may be related to the larger body size exhibited by females (156% heavier and 39% larger than males), as both length and weight had a significant effect on CTmax. The high energetic investment of females in offspring may also contribute to the differences recorded in thermal tolerance. Overall, organisms with a smaller body-size displayed a greater tolerance to elevated temperature, thus suggesting that smaller individuals may be positively selected in warmer environments. This selection may result in a reduction of size-at-maturity and shifts in sex ratio, given the sexual dimorphism in body size of shrimps. The thermal safety margin of P. varians was narrow (∼2.2 °C for males and ∼1.5 °C for females), revealing the vulnerability of this species to ocean warming and heatwaves.

Experimental evidence of contamination driven shrimp population dynamics: Susceptibility of populations to spatial isolation.

Contamination is likely to affect the composition of an ecological landscape, leading to the rupture of ecological connectivity among habitats (ecological fragmentation), which may impact on the distribution, persistence and abundance of populations. In the current study, different scenarios within a spatially heterogeneous landscape were simulated in the Heterogeneous Multi-Habitat Assay System (HeMHAS) to evaluate the potential effect that contamination (copper at 0.5 and 25 µg/L) might have on habitat selection by the estuarine shrimp Palaemon varians in combination with two other ecological factors: predator presence and food availability. As a result, P. varians detected and avoided copper; however, in the presence of the predation signal, shrimps shifted their response by moving to previously avoided regions, even if this resulted in a higher exposure to contamination. When encouraged to move towards environments with a high availability of food, a lower connectivity among the shrimp populations isolated by both contamination and predation risk simultaneously was evidenced, when compared to populations isolated only by the risk of predation. These results indicate that contamination might: (i) trigger avoidance in shrimps, (ii) prevent colonization of attractive foraging areas, (iii) enhance populations' isolation and (iv), make populations more susceptible to local extinction.

Functional and Stress Response Analysis of Heat Shock Proteins 40 and 90 of Giant River Prawn (Macrobrachium rosenbergii) under Temperature and Pathogenic Bacterial Exposure Stimuli.

The aims of this research were to perform molecular characterization and biofunctional analyses of giant river prawn Hsp40 and Hsp90 genes (Mr-hsp40 and Mr-hsp90) under various stress conditions. Comparisons of the nucleotide and amino acid sequences of Mr-hsp40 and Mr-hsp90 with those of other species showed the highest similarity scores with crustaceans. Under normal conditions, expression analysis using quantitative real-time RT-PCR (qRT-PCR) indicated that Mr-hsp40 was highly expressed in the gills and testis, and Mr-hsp90 expression was observed in all tissues, with the highest expression in the ovary. The expression patterns of Mr-hsp40 and Mr-hsp90 transcripts under Aeromonas hydrophila challenge and heat-cold shock conditions were examined in gills, the hepatopancreas and hemocytes, at 0, 3, 6, 12, 24, 48 and 96 h by qRT-PCR. Under bacterial challenge, Mr-hsp40 displayed variable expression patterns in all tissues examined during the tested periods. In contrast, upregulated expression of Mr-hsp90 was quickly observed from 3 to 12 h in the gills and hepatopancreas, whereas obviously significant upregulation of Mr-hsp90 was observed in hemocytes at 12-96 h. Under temperature shock conditions, upregulation of Mr-hsp40 expression was detected in all tested tissues, while Mr-hsp90 expression was quickly upregulated at 3-48 h in all tissues in response to 35 °C conditions, and conditions of 35 and 25 °C stimulated its expression in gills and the hepatopancreas at 12 and 48 h, respectively. Silencing analyses of these two genes were successfully conducted under normal, high-temperature (35 °C) and A. hydrophila infection conditions. Overall, knockdown of Mr-hsp40 and Mr-hsp90 effectively induced more rapid and higher mortality than in the PBS control and GFP induction groups in temperature and infectious treatments. Evidence from this study clearly demonstrated the significant functional roles of Mr-hsp40 and Mr-hsp90, which are crucially involved in cellular stress responses to both temperature and pathogenic bacterial stimuli.

Acclimation to cyclic hypoxia improves thermal tolerance and copper survival in the caridean shrimp Palaemon varians.

In response to the continuous variation of environmental parameters, species must be able to adjust their physiology to overcome stressful conditions, a process known as acclimatization. Numerous laboratory studies have been conducted to understand and describe the mechanisms of acclimation to one environmental stressor (e.g. cyclic hypoxia), but currently our understanding of how acclimation to one stressor can change tolerance to a subsequent stressor is limited. Here, in two different experiments, we used the shrimp Palaemon varians to test how, following 28-days acclimation to cyclic hypoxia (mimicking a cyclic hypoxic regime currently found in its natural habitat), critical thermal maximum (CTmax) and sensitivity to copper (Cu2+) exposure (30 mgL-1) changed in comparison to shrimp acclimated to normoxic conditions and then exposed to thermal stress or Cu2+. Acclimation to cyclic hypoxia improved both CTmax (~1 °C higher than controls) and survival to acute Cu2+ exposure (~30% higher than controls) and induced significant gene expression changes (i.e. up-regulation of heat shock protein 70 - HSP70, hypoxia inducible factor - HIF, phosphoenolpyruvate carboxykinase - PEPCK, glucose 6-P transporter - G6Pt, metallothionein - Mt, and down-regulation of hemocyanin - Hem) in animals acclimated to cyclic hypoxia. Our results demonstrate how acclimation to cyclic hypoxia improved tolerance to subsequent stressors, highlighting the complexity of predicting organismal performance in variable (i.e. where multiple parameters can simultaneously change during the day) environments.

Two genes related to reproductive development in the juvenile prawn, Macrobrachium nipponense: Molecular characterization and transcriptional response to nanoplastic exposure.

Plastic pollution in the form of nanoplastics poses a global threat to aquatic ecosystems and the organisms inhabiting them. However, few studies have been conducted on the effects of nanoplastic exposure on reproductive development in crustaceans. In order to address this issue, juvenile oriental river prawns (Macrobrachium nipponense) were exposed to different concentrations of 75-nm polystyrene nanoplastics (0, 5, 10, 20, 40 mg/L) for 28 days. In order to study the regulation of reproduction-related genes in the presence of nanoplastics, the Wee1 protein kinase gene (Wee1) and OTU domain ubiquitin aldehyde binding protein gene (OTUB) were selected. In this study, for the first time, the full-length cDNA of Mn-Wee1 and Mn-OTUB were cloned from M. nipponense. Homologous alignments revealed that Mn-Wee1 had a highly conserved function-critical sequence, and that Mn-OTUB was more closely related to OTUB1 than OTUB2. With increasing concentration of nanoplastics, the expression of both genes increased initially, then decreased. The inhibition of expression of Wee1 and OTUB occurred in 40 mg/L group, respectively. Analysis of the data also indicated that nanoplastic exposure might have differing effects on gene expression in M. nipponense male and female reproductive organs.

Salt transport by the gill Na+-K+-2Cl- symporter in palaemonid shrimps: exploring physiological, molecular and evolutionary landscapes.

Palaemonid shrimps inhabit osmotic niches from marine to continental waters. They hyper-regulate hemolymph osmolality and ionic concentrations in dilute media, hypo-regulating in concentrated media. Their gill epithelia express ion transporters like the Na+-K+-2Cl- symporter (NKCC) thought to play a role in salt secretion. To examine Cl- hypo-regulatory capability and phylogenetic correlations between gill NKCC mRNA levels and protein expression, we used palaemonids ranging from marine tide pools through estuaries (Palaemon) to coastal and continental fresh waters (Macrobrachium). We established the species' upper critical salinity limits (UL50) and short- (24 h) and long-term (120h) hypo-regulatory abilities at salinities of 80% of their UL50's (80%UL50). The Palaemon species exhibited the highest UL50's and greatest hypo-regulatory capabilities; among the Macrobrachium species, UL50's were higher in the diadromous than in the hololimnetic species. While basal transcript levels of gill NKCC mRNA were highest in P. pandaliformis, levels were unaffected by salinity or exposure time in all species. However, gill NKCC protein abundance increased after 120-h exposure at the 80%UL50 in all Macrobrachium species, except M. potiuna. Unexpectedly, hemolymph hyper-osmoregulatory capability in acclimatization media correlated with gill NKCC protein synthesis, while gill NKCC mRNA expression correlated with hemolymph hyper-Cl- regulation in Macrobrachium. These findings, together with the evolutionary history of osmoregulation in this shrimp clade, suggest a role for the gill NKCC symporter in both salt uptake and secretion. The evolution of NKCC protein expression responsiveness, unlike hemolymph hypo-regulation and NKCC mRNA expression, may have been driven by environmental salinity during niche radiation. SUMMARY STATEMENT: While mRNA expression of the gill Na+-K+-2Cl- symporter is unchanged during acclimation of palaemonid shrimps to saline media, protein expression is up regulated, revealing a role in chloride secretion.

Two new species of emTypton/em Costa, 1844 from tropical American waters, with taxonomic notes on emT. tortugae/em McClendon, 1911 and a new record of br /emT. granulosus/em Ayón-Parente, Hendrickx amp; Galvan-Villa, 2015 (Decapoda: Caridea: Palaemonidae).

Two new species of the palaemonid shrimp genus Typton Costa, 1844 are described based on material from Panama and Mexico. Both species are closely related to T. tortugae McClendon, 1911, a species originally described from the Dry Tortugas, off southern Florida, USA, and later scarcely recorded from other western Atlantic localities, from Bermuda to Mexico and Brazil. Some clarification and additional illustrations are provided for the type material of T. tortugae. Typton jonkayei sp. nov., is described based on material from fouling-encrusting communities dominated by sponges, growing on submerged roots of the red mangrove, Rhizhophora mangle L., in Bocas del Toro, Caribbean coast of Panama. This new species differs from T. tortugae in several morphological details, for instance, on the minor and major chelipeds (second pereiopods), telson, uropod, frontal margin and ambulatory pereiopods. Typton cousteaui sp. nov. is described based on a single ovigerous female dredged in the southern Gulf of California off Baja California Sur, Mexico, previously reported as T. tortugae. This new taxon seems to represent a true cryptic species with no significant morphological divergence from the allopatrically isolated T. tortugae, except for slight morphometric differences. In addition, T. granulosus Ayón-Parente, Hendrickx Galvan-Villa, 2015 is recorded from the Pacific coast of Panama, based on material collected in the Coiba Archipelago. Some taxonomic, distributional and ecological remarks are provided for T. granulosus and the closely related T. serratus Holthuis, 1951.

New data on Beurlenia araripensis Martins-Neto & Mezzalira, 1991, a lacustrine shrimp from Crato Formation, and its morphological variations based on the shape and the number of rostral spines.

Fossil freshwater carideans are very rare worldwide. Here, we present new taxonomic remarks about Beurlenia araripensis from the Early Cretaceous laminated limestones of the Crato Formation, Araripe Basin, northeastern Brazil. We analyzed five fossil samples, testing the morphological variations such as, rostrum with 5 to 14 supra-rostral spines and 2 to 3 sub-rostral spines, which appears as serrate for Caridea. This variation demonstrates a morphologic plasticity also seen in extant species of the group, such as those of the genera Macrobrachium and Palaemon.

MicroRNA transcriptome analysis of oriental river prawn Macrobrachium nipponense in responding to starvation stress.

Food deprivation or fasting is an important environmental factor, and a regular occurrence in both natural aquatic habitats and artificial ponds. However, the potential immunoregulatory mechanisms underlying starvation stress in crustaceans remain unclear. MicroRNAs (miRNAs) are a new class of non-coding RNAs that can regulate various biological processes, such as stress and immune responses. In the present work, miRNAs related to starvation stress responses and immune properties were identified and characterised in oriental river prawn Macrobrachium nipponense using high-throughput sequencing and bioinformatics analyses. Twelve small RNA libraries from hepatopancreas tissue were sequenced across four fasting stages lasting 0, 7, 14 or 21 days. In total, 550 miRNAs were identified including 198 putative novel miRNAs and 352 conserved miRNAs belonging to 57 families. Moreover, compared with expression levels at 0 days, 27, 27 and 43 miRNAs were differentially expressed (DE-miRNAs) at 7, 14 and 21 days, respectively. Among these, four DE-miRNAs (ame-miR-190-5p, dme-miR-307a-3p, hme-miR-2788-3p and novel_68) were co-expressed at all three timepoints. Furthermore, 661 target genes regulated by these DE-miRNAs were identified, and associated functional annotations were derived by GO enrichment and KEGG pathway analyses, which showed that most DE-miRNAs were mainly participated in metabolic processes and immune responses. Furthermore, 26 host DE-miRNAs potentially participated in interactions with white spot syndrome virus (WSSV) were identified by predicting and analysing target genes from WSSV. The further WSSV challenge under starvation stress showed that dme-miR-307a-3p played a part in the antiviral responses against WSSV. Our results demonstrate that dme-miR-307a-3p may play vital regulatory roles in responding to starvation stress and WSSV infection. The findings contribute new insight into the molecular mechanisms associated with immune responses to environmental stress in crustaceans.

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 analysis of lethality in response to RNA interference of the oriental river prawn (Macrobrachium nipponense).

A previous study identified slow-tonic S2 tropomyosin and slow tropomyosin isoform as sex-related genes in Macrobrachium nipponense. Their functions were analyzed using RNA interference. However, more than half of the specimens died approximately 8-12 h after injection of the respective double-stranded RNAs (dsRNAs), and HE staining indicated that the heart and gills were the most likely tissues responsible for the resultant deaths. In the current study, we conducted a comparative transcriptomic study of the gills and hearts of M. nipponense to identify potential target genes associated with acute death after dsRNA injection. A total of 68,772 annotated unigenes were generated. In the heart, differentially expressed genes (DEGs) were mainly enriched in glycolysis/gluconeogenesis and oxidative phosphorylation, while the most relevant pathways in the gills were lysosome, phagosome, and peroxisome. Ten DEGs were screened out and analyzed under lethal hypoxic stress. Among these, fructose 1, 6-biphosphate-aldolase (FBA), glyceraldehyde 3-phosphate dehydrogenase (GDPDH), alcohol dehydrogenase class-3 (ADC3), ATP-synthase subunit 9 (ATPS9), and acid ceramidase-like (ACL) were all differentially expressed under hypoxic conditions. This study shed light on the lethal mechanism caused by interference with tropomyosin genes in M. nipponense, and identifies the related pathways and key genes that could help to improve stress resistance and tolerance in M. nipponense.

Molecular characterization and expression dynamics of three key genes in the PI3K-AKT pathway reveal its involvement in the immunotoxicological responses of the giant river prawn Macrobrachium rosenbergii to acute ammonia and nitrite stress.

Ammonia nitrogen and nitrite are two common forms of environmental toxicants for aquatic organisms including crustaceans. The PI3K-AKT pathway is an important intracellular signaling pathway related to cellular stress response, but involvement of this pathway in the immunotoxicological response of decapod crustaceans to aquatic toxicants such as ammonia nitrogen and nitrite still remains enigmatic. In this study, based on transcriptome mining and molecular cloning techniques, three key genes (named as MrPI3K, MrAKT and MrFoxO) in the PI3K-AKT signaling pathway were identified from the giant river prawn Macrobrachium rosenbergii. Sequence homology and phylogenetic analysis revealed that all the three genes harbored signature sequences of corresponding protein families, and shared high levels of similarities with their respective homologs from other species. MrPI3K, MrAKT and MrFoxO all displayed ubiquitous tissue distribution profiles, but their expression levels varied to a great extend among different tissues and between sexes. Following exposure to nitrite (20 mg/L nitrite-N) or ammonia (25 mg/L total ammonia-N) stresses for 24 h and 48 h, the three genes all responded by altering their expression levels at different time points, but they didn't show uniform expression patterns following these stresses, indicating the diversified roles of these genes in different tissues and the complexity of this signaling pathway. Remarkably, MrPI3K and MrAKT were induced only in the hemocytes and intestine, respectively, indicating their specific roles in these organs. Our study demonstrated the potential utility of these genes as biomarkers of acute ammonia or nitrite toxicity in prawns, and also provided evidence that the PI3K-AKT pathway is involved in the immunotoxicological responses to nitrite and ammonia stress in M. rosenbergii.

Cadmium-induced oxidative stress, metabolic dysfunction and metal bioaccumulation in adult palaemonid shrimp Palaemon macrodactylus (Rathbun, 1902).

The Palaemonid shrimp Palaemon macrodactylus is widely distributed in coastal areas and estuaries which are easily contaminated by various pollutants. However, the responses of this species to environmental toxicants are not well described. In the present study, adult individuals of P. macrodactylus were exposed to gradient concentrations of Cadmium (Cd) to evaluate its acute toxic effects, including bioaccumulation, induced oxidative stress and changed energy metabolism in this species. The medium lethal concentration (LC50) of Cd at 24 h, 48 h, 72 h, and 96 h were 2.60, 0.88, 0.49 and 0.37 mg/L, respectively. Cd bioaccumulations in tissues of shrimp increased in a concentration-dependent manner, and higher concentration (50% 96 h-LC50, 0.185 mg/L) of Cd exposure led to a maximum increase of Cd concentration by 14.8, 145.5 and 15.8 folds in gill, hepatopancreas and abdominal muscle. Cd exposure caused a significant inhibition on the activity of catalase (CAT), and total superoxide dismutase (T-SOD), decrease in the total antioxidant capacity (T-AOC), and an increase of malonadehyde (MDA) content, which indicated a damage to the antioxidant system of shrimp. Meanwhile, Cd exposure also led to a significant up-regulation in the expression level of metallothionein gene (MT), and down-regulations at the mRNA level of heat shock protein 70 (HSP70) and CAT. Moreover, Cd exposure significantly inhibited the oxygen consumption rate (22%), and increased the ammonia excretion rate (43%), hence lead to a significant decrease of the O:N ratio (45%) in shrimp. The results indicated that Cd exposure could induce obvious oxidative stress, energy metabolic dysfunction and bioaccumulation of Cd in P. macrodactylus. The data obtained from the present study would provide useful information for further understanding on the toxicological mechanism of Cd to crustaceans in coastal areas and estuaries.

Do plasticity in gene expression and physiological responses in Palaemonid prawns facilitate adaptive response to different osmotic challenges?

Integrating physiological and genomic approaches in a comparative framework offers excellent opportunity to investigate the underlying mechanisms for acclimation to specific challenges. The present study was conducted on three different prawn species (inhabitants of different salinity environments) of the genus Macrobrachium (M. australiense, M. tolmerum and M. novaehollandiae) to investigate the salinity induced changes in expression patterns of 10 candidate genes in the gill tissue (that previously had been inferred to play important functional roles in acclimation and adaptation to freshwater environments), and hemolymph osmolality. The prawn individuals were maintained in laboratory condition under three different salinity levels (0‰, 6‰ and 12‰) for 28 days using 6‰ as the control. All of the genes studied, showed salinity induced differential expression patterns. Genes with more important functional roles under low ionic conditions (i.e. Claudin, Na+/H+exchanger, V-type H+-ATPase and UNT2) showed 2.5 to 6 fold higher expression at 0‰ compared with at higher salinities (6‰ and 12‰) but no significant differences (p > 0.05) were observed between 6‰ and 12‰ for the same genes. In contrast, 1.5 to 4 fold higher expression levels were observed at 6‰ and 12‰ for genes that have important roles in mediating salinity tolerance (i.e., Na+/K+-ATPase, Na+/K+/2Cl-Co-transporter, Diuretic Hormone, Crustacean Hyperglycaemic Hormone and UNT1). The osmotic stress response gene, Calreticulin, showed significant differences (p < 0.05) in expression between different salinity comparisons. Hemolymph osmolality also was impacted in all three species with a strong correlation evident between hemolymph osmolality and expression of genes influencing this trait. Findings indicate an important role of plasticity that facilitates rapid acclimation to changing salinity levels.