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Parabens are preservatives widely used in personal care products, pharmaceuticals, and foodstuffs. However, they are still unregulated chemical compounds. Given their extensive use and presence in different environmental compartments, parabens can adversely affect animal health. Thus, the current study aimed to summarize and critically analyze the bioaccumulation and ecotoxicity of parabens in aquatic species. Studies have been mostly conducted in laboratory conditions (75%), using mainly fish and crustaceans. Field studies were carried out across 128 sampling sites in six countries. Paraben bioaccumulation was predominantly detected in fish muscle, liver, brain, gills, ovary, and testes. Among the parent parabens, methylparaben (MeP), ethylparaben (EtP), and propylparaben (PrP) have been detected frequently and more abundantly in tissues of marine and freshwater specimens, as well as the metabolite 4-hydroxybenzoic acid (4-HB). Parabens can induce lethal and sublethal effects on aquatic organisms, such as oxidative stress, endocrine disruption, neurotoxicity, behavioral changes, reproductive impairment, and developmental abnormalities. The toxicity of parabens varied according to species, taxonomic group, developmental stage, exposure time, and concentrations tested. This study highlights the potential bioaccumulation and ecotoxicological impacts of parabens and their metabolites on aquatic invertebrates and vertebrates. Additionally, future research recommendations are provided to evaluate the environmental risks posed by paraben contamination more effectively., 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 Ltd. All rights reserved.)

Galectins are a member of the β-galactoside binding protein family, which play a pivotal role in the immune defense of vertebrates as a pattern recognition receptor and occupy an important position in the innate immune system of invertebrates. The study of galectins in aquatic organisms has only recently emerged. Galectins in aquatic animals exhibit agglutination activity toward bacteria, inhibit bacterial growth, and enhance phagocytosis of immune cells. Additionally, some galectins contribute to the antiviral immune defenses of aquatic animals. This review aims to review recent advancements in the antimicrobial mechanisms, molecular structures, and evolution of galectins from fish, mollusks, and crustaceans. The antimicrobial galectins, as crucial components in the innate immune defense, pave new avenues for developing innovative disease control strategies in aquaculture.

Evolutionary innovations in chemical secretion-such as the production of secondary metabolites, pheromones, and toxins-profoundly impact ecological interactions across a broad diversity of life. These secretory innovations may involve a "legacy-plus-innovation" mode of evolution, whereby new biochemical pathways are integrated with conserved secretory processes to create novel products. Among secretory innovations, bioluminescence is important because it evolved convergently many times to influence predator-prey interactions, while often producing courtship signals linked to increased rates of speciation. However, whether or not deeply conserved secretory genes are used in secretory bioluminescence remains unexplored. Here, we show that in the ostracod Vargula tsujii, the evolutionary novel c-luciferase gene is co-expressed with many conserved genes, including those related to toxin production and high-output protein secretion. Our results demonstrate that the legacy-plus-innovation mode of secretory evolution, previously applied to sensory modalities of olfaction, gustation, and nociception, also encompasses light-producing signals generated by bioluminescent secretions. This extension broadens the paradigm of secretory diversification to include not only chemical signals but also bioluminescent light as an important medium of ecological interaction and evolutionary innovation., Competing Interests: Conflict of Interest The authors declare no competing interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)

Heavy metals pose significant threats to ecosystems and human health due to their persistence and bioaccumulation. In Thailand, rapid industrialization, extensive agriculture, and urban development have exacerbated heavy metal pollution in both aquatic and terrestrial ecosystems. This systematic review, conducted according to PRISMA guidelines, evaluates study designs and methodologies to assess heavy metal bioaccumulation in Thai fauna, with a focus on ecological and health impacts. The review reveals that fish, particularly from families like Cyprinidae and Cichlidae, account for 42.11 % of studies, with species such as swamp eel, Henicorhynchus siamensis, Arius maculatus, Osteogeneiosus militaris, Puntioplites proctozystron, and Channa striata showing significant bioaccumulation. Molluscs (31.58 %), including Tegillarca granosa and Filopaludina martensi, serve as critical bioindicators of aquatic pollution due to their filter-feeding habits. Amphibians and crustaceans, like Fejervarya limnocharis and Fenneropenaeus merguiensis, also demonstrate vulnerability to heavy metal contamination. Key contamination hotspots include urban waterways in Bangkok, industrial discharges in Songkhla Lake, and mining sites in Loei Province, highlighting widespread environmental and health impacts. Despite extensive research, gaps remain, particularly concerning benthic scavengers and detritivores, which are vital for ecosystem functions. The review underscores the need for targeted monitoring and mitigation, including stricter regulations on industrial discharges, improved waste treatment, and better management of agricultural runoff. While metals like cadmium (Cd), lead (Pb), copper (Cu), and zinc (Zn) are well-studied, further research on less-examined metals and species-specific bioaccumulation patterns is crucial to enhancing environmental management, supporting biodiversity conservation, and improving ecosystem resilience in Thailand., 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 The Authors. Published by Elsevier Inc. All rights reserved.)

Pharmaceuticals (PhACs) are increasingly detected in aquatic ecosystems, yet their effects on biota remain largely unknown. The environmentally relevant concentrations of many PhACs may not result in individual-level responses, like mortality or growth inhibition, traditional toxicity endpoints. However, this doesn't imply the absence of negative effects on biota. Metabolomics offers a more sensitive approach, detecting responses at molecular and cellular levels and providing mechanistic understanding of adverse effects. We evaluated bioaccumulation and metabolic alterations in a benthic ostracod, Heterocypris incongruens, exposed to a mixture of five PhACs (carbamazepine, tiapride, tolperisone, propranolol and amlodipine) at environmentally relevant concentrations for 7 days using liquid chromatography coupled with mass spectrometry. The selection of PhACs was based, among other factors, on risk quotient values determined using toxicological data available in the literature and concentrations of PhACs quantified in our previous research in the sediments of the Odra River estuary. This represents a novel approach to PhACs selection for metabolomic studies that considers strictly quantitative data. Amlodipine and tolperisone exhibited the highest bioaccumulation. Significant impacts were observed in Alanine, aspartate and glutamate metabolism, Starch and sucrose metabolism, Arginine biosynthesis, Histidine metabolism, Tryptophan metabolism, Glycerophospholipid metabolism, and Glutathione metabolism pathways. Most of the below-individual-level responses were likely nonspecific and related to dysregulation in energy metabolism and oxidative stress response. Additionally, some pharmaceutical-specific responses were also observed. Therefore, untargeted metabolomics can be used to detect metabolic changes resulting from environmentally relevant concentrations of PhACs in aquatic ecosystems and to understand their underlying mechanism., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Joanna Giebultowicz reports financial support was provided by National Science Centre Poland. If there are other authors, they 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.)

Crustaceans played a major role in establishing that gamma-aminobutyric acid (GABA) functioned as an inhibitory transmitter compound. In fact, it is now widely accepted that GABA is the major inhibitory transmitter compound in all animal species where it has been examined. The story of its acceptance as a neurotransmitter, however, is more interesting than that. GABA was first isolated from mammalian brains by three laboratories in 1950. Great excitement surrounded this discovery, and many laboratories began exploring its function. This excitement peaked at two large international congresses in the United States at the end of the first decade of study, where a consensus of major figures in the field was that GABA was not a transmitter compound. How could this have happened?, Competing Interests: Declaration of Competing Interest The authors declare that the review was written and compiled in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The neuroendocrine system of crustaceans is complex and regulates many processes, such as development, growth, reproduction, osmoregulation, behavior, and metabolism. Once stimulated, crustaceans' neuroendocrine tissues modulate the release of monoamines, ecdysteroids, and neuropeptides that can act as hormones or neurotransmitters. Over a few decades, research has unraveled some mechanisms governing these processes, substantially contributing to understanding crustacean physiology. More aspects of crustacean neuroendocrinology are being comprehended with molecular biology, transcriptome, and genomics analyses. Hence, these studies will also significantly enhance the ability to cultivate decapods, such as crabs and shrimps, used as human food sources. In this review, current knowledge on crustacean endocrinology is updated with new findings about crustacean hormones, focusing mainly on the main neuroendocrine organs and their hormones and the effects of these molecules regulating metabolism, growth, reproduction, and color adaptation. New evidence about vertebrate-type hormones found in crustaceans is included and discussed. Finally, this review may assist in understanding how the emerging chemicals of environmental concern can potentially impair and disrupt crustacean's endocrine functions and their physiology., Competing Interests: Declaration of competing interest As the corresponding author, I am sending the manuscript “A brief and updated introduction to the neuroendocrine system of crustaceans” in order to submit for publication on the Special Issue “Hormonal Regulation and Disruption in Invertebrates” in the Journal of Molecular and Cellular Endocrinology. In the name of all authors, I certify that this manuscript is not under consideration by any other journal and that all the authors have read the manuscript and agreed to have their names listed as authors. I also certify that all the authors collaborated in the development of this research work. Finally, there is no potential conflict of interest including any financial, personal or other relationships with other people or organizations related to this manuscript., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Marine pollution caused by heavy metals has emerged as a significant environmental concern, garnering increased attention in recent years. The accumulation of heavy metals in the tissues of marine organisms poses substantial threats to both marine ecosystems and human populations that rely on seafood as a primary food source. Fish and crustaceans are effective biomonitors for assessing heavy metal contamination in aquatic environments. In this study, we determined the concentrations of several heavy metals, including cadmium (Cd), lead (Pb), nickel (Ni), mercury (Hg), and tin (Sn), in four fish species (Mugil cephalus, Mugil capito, L. aurata, and Morone labrax) and five crustacean species (S. rivulatus, Cerastoderma glaucum, Paratapes undulatus, R. decussatus, Callinectes sapidus, and Metapenaeus Stebbingi) from Temsah Lake during both winter and summer seasons. To evaluate the potential ecological and health risks associated with consuming these fish and crustacean species, we calculated the metal pollution index (MPI), weekly intake (EWI), target hazard quotient (THQ), and carcinogenic risk (CR) values. The results revealed a noticeable increase in metal levels during the summer compared to winter in the studied samples. Moreover, the concentration of heavy metals in the muscles of the species generally exceeded those in the liver and gills. The MPI values indicated that Morone labrax exhibited the highest values during winter, while L. aurata showed the highest values during summer. Mugil cephalus demonstrated the lowest MPI values in both seasons. The EWI values for the studied metals were found to be lower than the corresponding tolerable weekly intake (TWI) values. Additionally, under average exposure conditions, the THQ and HI data were generally below one for most study species in the area. The calculated CR values for investigated metals in the studied species indicated acceptable carcinogenic risk levels. Therefore, this suggests that consuming studied species within Temsah lake does not present any potential health hazards for consumers., (© 2024. The Author(s).)

Background: Hydrocarbon pollution stemming from petrochemical activities is a significant global environmental concern. Bioremediation, employing microbial chitinase-based bioproducts to detoxify or remove contaminants, presents an intriguing solution for addressing hydrocarbon pollution. Chitooligosaccharides, a product of chitin degradation by chitinase enzymes, emerge as key components in this process. Utilizing chitinaceous wastes as a cost-effective substrate, microbial chitinase can be harnessed to produce Chitooligosaccharides. This investigation explores two strategies to enhance chitinase productivity, firstly, statistical optimization by the Plackett Burman design approach to  evaluating the influence of individual physical and chemical parameters on chitinase production, Followed by  response surface methodology (RSM) which delvs  into the interactions among these factors to optimize chitinase production. Second, to further boost chitinase production, we employed heterologous expression of the chitinase-encoding gene in E. coli BL21(DE3) using a suitable vector. Enhancing chitinase activity not only boosts productivity but also augments the production of Chitooligosaccharides, which are found to be used as emulsifiers., Results: In this study, we focused on optimizing the production of chitinase A from S. marcescens using the Plackett Burman design and response surface methods. This approach led to achieving a maximum activity of 78.65 U/mL. Subsequently, we cloned and expressed the gene responsible for chitinase A in E. coli BL21(DE3). The gene sequence, named SmChiA, spans 1692 base pairs, encoding 563 amino acids with a molecular weight of approximately 58 kDa. This sequence has been deposited in the NCBI GenBank under the accession number "OR643436". The purified recombinant chitinase exhibited a remarkable activity of 228.085 U/mL, with optimal conditions at a pH of 5.5 and a temperature of 65 °C. This activity was 2.9 times higher than that of the optimized enzyme. We then employed the recombinant chitinase A to effectively hydrolyze shrimp waste, yielding chitooligosaccharides (COS) at a rate of 33% of the substrate. The structure of the COS was confirmed through NMR and mass spectrometry analyses. Moreover, the COS demonstrated its utility by forming stable emulsions with various hydrocarbons. Its emulsification index remained stable across a wide range of salinity, pH, and temperature conditions. We further observed that the COS facilitated the recovery of motor oil, burned motor oil, and aniline from polluted sand. Gravimetric assessment of residual hydrocarbons showed a correlation with FTIR analyses, indicating the efficacy of COS in remediation efforts., Conclusions: The recombinant chitinase holds significant promise for the biological conversion of chitinaceous wastes into chitooligosaccharides (COS), which proved its potential in bioremediation efforts targeting hydrocarbon-contaminated sand., (© 2024. The Author(s).)

Genes from ancient families are sometimes involved in the convergent evolutionary origins of similar traits, even across vast phylogenetic distances. Sulfotransferases are an ancient family of enzymes that transfer sulfate from a donor to a wide variety of substrates, including probable roles in some bioluminescence systems. Here, we demonstrate multiple sulfotransferases, highly expressed in light organs of the bioluminescent ostracod Vargula tsujii , transfer sulfate in vitro to the luciferin substrate, vargulin. We find luciferin sulfotransferases (LSTs) of ostracods are not orthologous to known LSTs of fireflies or sea pansies; animals with distinct and convergently evolved bioluminescence systems compared to ostracods. Therefore, distantly related sulfotransferases were independently recruited at least three times, leading to parallel evolution of luciferin metabolism in three highly diverged organisms. Reuse of homologous genes is surprising in these bioluminescence systems because the other components, including luciferins and luciferases, are completely distinct. Whether convergently evolved traits incorporate ancient genes with similar functions or instead use distinct, often newer, genes may be constrained by how many genetic solutions exist for a particular function. When fewer solutions exist, as in genetic sulfation of small molecules, evolution may be more constrained to use the same genes time and again.

C-type lectins (CTLs) function as pattern recognition receptors (PRRs) by recognizing invading microorganisms, thereby triggering downstream immune events against infected pathogens. In this study, a novel CTL containing a low-density lipoprotein receptor class A (LDLa) domain was obtained from Litopenaeus vannamei, designed as LvLDLalec. Stimulation by the bacterial pathogen Vibrio anguillarum (V. anguillarum) resulted in remarkable up-regulation of LvLDLalec, as well as release of LvLDLalec into hemolymph. The rLvLDLalec protein possessed broad-spectrum bacterial binding and agglutinating activities, as well as hemocyte attachment ability. Importantly, LvLDLalec facilitated the bacterial clearance in shrimp hemolymph and protected shrimp from bacterial infection. Further studies revealed that LvLDLalec promoted hemocytes phagocytosis against V. anguillarum and lysosomes were involved in the process. Meanwhile, LvLDLalec participated in humoral immunity through activating and inducing nuclear translocation of Dorsal to regulate phagocytosis-related genes and antimicrobial peptides (AMPs) genes, thereby accelerated the removal of invading pathogens in vivo and improved the survival rate of L. vannamei. These results unveil that LvLDLalec serves as a PRR participate in cellular and humoral immunity exerting opsonin activity to play vital roles in the immune regulatory system of L. vannamei., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Green biorefinery process was conducted to extract α-chitin and high-value co-products from shrimp shell waste through microbial fermentation using mature coconut water (MCW) as a sole nutrient source. Symbiotic co-lactic acid fermentation (Co-LAF) by Lactobacillus plantarum and Streptococcus thermophilus produced higher levels of lactic acid (LA) and protease activity than their mono-cultures, which led to greater demineralization (DM) and deproteinization (DP) of shrimp shell powder (SSP). After optimizing Co-LAF through Response Surface Methodology and successive fermentation by an acid-active proteolytic fungus Rhizopus oligosporus, the highest DM of 94.0 ± 0.91 % and DP of 86.7 ± 0.1 % were achieved. Based on FT-IR, XRD, and SEM analysis, the bio-extracted chitin had similar structural characteristics to commercial α-chitin but with better quality. These strategies not only contribute to environmentally-friendly and cost-effective extraction of α-chitin (303 ± 18 mg/g-SSP), but also co-produce LA (57.18 ± 0.89 g/L), acid protease (4.33 ± 0.5 U/mL), bio-calcium (277 ± 12 mg-CaSO 4 /g-SSP), protein hydrolysate (268 ± 5 mg/g-SSP), and pigments (28.78 ± 1.56 µg/g-SSP)., 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 © 2023 Elsevier Ltd. All rights reserved.)

The unscientific disposal of the most abundant crustacean wastes, especially those derived from marine sources, affects both the economy and the environment. Strategic waste collection and management is the need of the hour. Sustainable valorization approaches have played a crucial role in solving those issues as well as generating wealth from waste. The shellfishery wastes are rich in valuable bioactive compounds such as chitin, chitosan, minerals, carotenoids, lipids, and other amino acid derivatives. These value-added components possessed pleiotropic applications in different sectors viz., food, nutraceutical, cosmeceutical, agro-industrial, healthcare, and pharmaceutical sectors. The manuscript covers the recent status, scope of shellfishery management, and different bioactive compounds obtained from crustacean wastes. In addition, both sustainable and conventional routes of valorization approaches were discussed with their merits and demerits along with their combinations. The utilization of nano and microtechnology was also included in the discussion, as they have become prominent research areas in recent years. More importantly, the future perspectives of crustacean waste management and other potential valorization approaches that can be implemented on a large 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 © 2023 Elsevier B.V. All rights reserved.)

Ferroptosis, characterized by iron-dependent cell death, has recently emerged as a critical defense mechanism against microbial infections. The present study aims to investigate the involvement of exosomes in the induction of ferroptosis and the inhibition of bacterial infection in crustaceans. Our findings provide compelling evidence for the pivotal role of exosomes in the immune response of crustaceans, wherein they facilitate intracellular iron accumulation and activate the ferroptotic pathways. Using RNA-seq and bioinformatic analysis, we demonstrate that cytochrome P450 (CYP) can effectively trigger ferroptosis. Moreover, by conducting an analysis of exosome cargo proteins, we have identified the participation of six-transmembrane epithelial antigen of prostate 4 in the regulation of hemocyte ferroptotic sensitivity. Subsequent functional investigations unveil that six-transmembrane epithelial antigen of prostate 4 enhances cellular Fe 2+ levels, thereby triggering Fenton reactions and accelerating CYP-mediated lipid peroxidation, ultimately culminating in ferroptotic cell death. Additionally, the Fe 2+ -dependent CYP catalyzes the conversion of arachidonic acid into 20-hydroxyeicosatetraenoic acid, which activates the peroxisome proliferator-activated receptor. Consequently, the downstream target of peroxisome proliferator-activated receptor, cluster of differentiation 36, promotes intracellular fatty acid accumulation, lipid peroxidation, and ferroptosis. These significant findings shed light on the immune defense mechanisms employed by crustaceans and provide potential strategies for combating bacterial infections in this species., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

The light-dark cycle significantly impacts the growth and development of animals. Mantis shrimps (Oratosquilla oratoria) receive light through their complex photoreceptors. To reveal the adaptive expression mechanism of the mantis shrimp induced in a dark environment, we performed comparative transcriptome analysis with O. oratoria cultured in a light environment (Oo-L) as the control group and O. oratoria cultured in a dark environment (Oo-D) as the experimental group. In the screening of differentially expressed genes (DEGs) between the Oo-L and Oo-D groups, a total of 88 DEGs with |log2FC| > 1 and FDR < 0.05 were identified, of which 78 were upregulated and 10 were downregulated. Then, FBP1 and Pepck were downregulated in the gluconeogenesis pathway, and MKNK2 was upregulated in the MAPK classical pathway, which promoted cell proliferation and differentiation, indicating that the activity of mantis shrimp was slowed and the metabolic rate decreases in the dark environment. As a result, the energy was saved for its growth and development. At the same time, we performed gene set enrichment analysis (GSEA) on all DEGs. In the KEGG pathway analysis, each metabolic pathway in the dark environment showed a slowing trend. GO was enriched in biological processes such as eye development, sensory perception and sensory organ development. The study showed that mantis shrimp slowed down metabolism in the dark, while the role of sensory organs prominent. It provides important information for further understanding the energy metabolism and has great significance to study the physiology of mantis shrimp in dark environment., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Predators in aquatic environments can be exposed to microplastics (MPs) directly through water and indirectly through prey. Laboratory experiments were conducted to study the potential of MP trophic transfer in Baltic Sea littoral food chains of different lengths. The longest studied food chain had three trophic levels: zooplankton, chameleon shrimp (Praunus flexuosus) and rockpool prawn (Palaemon elegans). 10 μm fluorescence microspheres were used as tracer MP particles and MP ingestion was verified with epifluorescence microscopy. Transfer of MPs occurred up to both second and third trophic level. The number of ingested microspheres in both chameleon shrimp and rockpool prawn was higher when the animals were exposed through pre-exposed prey in comparison to direct exposure through the water. The results show that trophic transfer may be an important pathway of and increase the microplastic exposure for some animals at higher trophic levels in highly polluted areas., 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 © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

For several decades, researchers have been using protein-fragment complementation assay (PCA) approaches for biosensing to study protein-protein interaction for a variety of aims, including viral infection, cellular apoptosis, G protein coupled receptor (GPCR) signaling, drug and substrate screening, and protein aggregation and protein editing by CRISPR/Cas9. As a reporter, NanoLuc (NLuc), a smaller and the brightest engineered luciferase derived from deep-sea shrimp Oplophorus gracilirostris , has been found to have many benefits over other luminescent enzymes in PCA. Inspired by the split green fluorescent protein (GFP) and its β-barrel structure, two split NLuc consisting of peptide fragments have been reported including the binary and ternary NLuc systems. NanoBiT® (large fragment + peptide) has been used extensively. In contrast, tripart split NLuc (large fragment + 2 peptides) has been applied and hardly used, while it has some advantages over NanoBiT in some studies. Nevertheless, tripart NLuc has some drawbacks and challenges to overcome but has several potential characteristics to become a multifunctional and powerful tool. In this review, several aspects of tripart NLuc are studied and a brief comparison with NanoBiT® is given.

This research aims to introduce a low-cost, non-commercial culture medium and optimize the operating conditions for biological chitin extraction from green tiger shrimp waste in the Persian Gulf zone. For this purpose, the two most commonly used microorganisms, Bacillus licheniformis and Lactobacillus plantarum, were obtained to deproteinize and demineralize the shrimp shells within both culture mediums using a successive two-stage process. It was found that the proposed non-commercial culture medium was more efficient than the purchased and ready-to-use commercial medium and increased deproteinization and demineralization efficiency by 9 % and 11 %, respectively. According to the optimization, which was performed using a response surface methodology based on a central composite design, the demineralization model is more complicated than the deproteinization model. The presented model predicted deproteinization and demineralization yields with good accuracy. The FTIR results revealed that shrimp shells and chitin have similar main functional groups, while the degree of acetylation of the extracted chitin was 62.26 %. SEM results illustrated the formation of microfibrils and the chitin structure's porosity. The XRD data showed that the crystallinity index of chitin was 93.9 %. Besides, the thermal stability of the extracted chitin, with a maximum degradation temperature of 380 °C is comparable with the literature data., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Shahriar Osfouri reports administrative support was provided by Persian Gulf University., (Copyright © 2023 Elsevier B.V. All rights reserved.)

As the main allergenic food, shrimp can trigger allergic reactions in various degrees. In this study, arginine kinase (AK) was identified as an allergen in Oratosquilla oratoria by LC-MS/MS. The open reading frame of AK was obtained, which included 356 amino acids, and recombinant AK (rAK) was expressed in Escherichia coli . The results of immunological analysis and circular dichroism showed that rAK displayed similar IgG-/IgE-binding activity and structure as native AK. Besides, five IgE linear epitopes of AK were verified by serological analysis, on the basis of which an epitope-deleted derivative was obtained and named as mAK-L. It has been shown that mAK-L displayed hypo-immunoreactivity compared to rAK, and the contents of secondary structures were different. In conclusion, these discoveries enrich the overall understanding of crustacean allergens and epitopes and set the foundations for food allergy diagnosis and immunotherapy.

The immune deficiency (IMD) pathway is critical for elevating host immunity in both insects and crustaceans. The IMD pathway activation in insects is mediated by peptidoglycan recognition proteins, which do not exist in crustaceans, suggesting a previously unidentified mechanism involved in crustacean IMD pathway activation. In this study, we identified a Marsupenaeus japonicus B class type III scavenger receptor, SRB2, as a receptor for activation of the IMD pathway. SRB2 is up-regulated upon bacterial challenge, while its depletion exacerbates bacterial proliferation and shrimp mortality via abolishing the expression of antimicrobial peptides. The extracellular domain of SRB2 recognizes bacterial lipopolysaccharide (LPS), while its C-terminal intracellular region containing a cryptic RHIM-like motif interacts with IMD, and activates the pathway by promoting nuclear translocation of RELISH. Overexpressing shrimp SRB2 in Drosophila melanogaster S2 cells potentiates LPS-induced IMD pathway activation and diptericin expression. These results unveil a previously unrecognized SRB2-IMD axis responsible for antimicrobial peptide induction and restriction of bacterial infection in crustaceans and provide evidence of biological diversity of IMD signaling in animals. A better understanding of the innate immunity of crustaceans will permit the optimization of prevention and treatment strategies against the arising shrimp diseases.

Crustacean female sex hormone (CFSH) is believed to regulate the development of female-related phenotypes in crustaceans. However, its role in gonadal development has been understudied. This study identified a CFSH gene, Lvit-CFSH1b, in the peppermint shrimp Lysmata vittata, a protandric simultaneous hermaphroditism (PSH) species. Lvit-CFSH1b is only expressed in the eyestalk ganglion. qRT-PCR showed that the expression level of Lvit-CFSH1b significantly increased with the gonad development from stage I to III (male phase) and decreased at stage IV (euhermaphrodite phase). Gene knockdown of Lvit-CFSH1b resulted in retardation of female phenotypes and stimulated the development of male phenotypes. At the same time, ovarian development was inhibited, and spermatogenesis was promoted. In addition, injection of rCFSH1b increased ovarian expression of vitellogenin (Lvit-Vg) and hepatopancreas expression of vitellogenin receptor (Lvit-VgR), while suppressing the expressions of insulin-like androgenic gland hormones (Lvit-IAG1 and Lvit-IAG2) in androgenic glands. The addition of rCFSH1b induced the in vitro expression of Lvit-Vg in ovarian and Lvit-VgR in hepatopancreas explants. In conclusion, this study provides convincing evidence that CFSH expedites the feminization process and impedes masculinization by inhibiting IAG in hermaphroditic crustaceans., 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 © 2023. Published by Elsevier B.V.)

One-step fermentation, inoculated with Lactobacillus fermentum ( L. fermentum ) in shrimp by-products, was carried out to obtain chitin and flavor protein hydrolysates at the same time. The fermentation conditions were optimized using response surface methodology, resulting in chitin with a demineralization rate of 89.48%, a deproteinization rate of 85.11%, and a chitin yield of 16.3%. The surface of chitin after fermentation was shown to be not dense, and there were a lot of pores. According to Fourier transform infrared spectroscopy and X-ray diffraction patterns, the fermented chitin belonged to α-chitin. More than 60 volatiles were identified from the fermentation broth after chitin extraction using gas chromatography-ion transfer spectrometry analysis. L. fermentum fermentation decreased the intensities of volatile compounds related to unsaturated fatty acid oxidation or amino acid deamination. By contrast, much more pleasant flavors related to fruity and roasted aroma were all enhanced in the fermentation broth. Our results suggest an efficient one-step fermentation technique to recover chitin and to increase aroma and flavor constituents from shrimp by-products.

Selective breeding for acute hepatopancreatic necrosis disease (AHPND) resistant shrimp is an effective way to deal with heavy losses to shrimp aquaculture caused by AHPND. However, knowledge about the molecular mechanism of susceptibility or resistance to AHPND is very limited. We herein performed a comparative transcriptomic analysis of gill tissue between AHPND susceptible and resistant families of the white Pacific shrimp Litopenaeus vannamei during Vibrio parahaemolyticus (VP AHPND ) infection. A total of 5,013 genes that were differentially expressed between the two families at 0 and 6 h post-infection, and 1,124 DEGs were shared for both two time points. Both GO and KEGG analyses in each or two time point's comparisons showed DEGs involved in endocytosis, protein synthesis and cell inflammation were significantly enriched. Several immune DEGs including PRRs, antioxidants and AMPs were also identified. The susceptible shrimp showed enhanced endocytosis, higher aminoacyl-tRNA ligase activity and occurrence of inflammatory response, while the resistant shrimp had much more strong ability in ribosome biogenesis, antioxidant activity and pathogen recognition and clearance. These genes and processes were mostly associated with mTORC1 signaling pathway, which could reflect differences in cell growth, metabolism and immune response between the two families. Our findings indicate a close link between mTORC1 signaling-related genes and Vibrio -resistance phenotype of shrimp, and provide new clues for further research on resistance strategy of shrimp to AHPND., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Liu, Yu, Li, Sun and Li.)

Early Mortality Syndrome (EMS) has been a major problem for shrimp aquaculture in Southeast Asia due to its epizootic prevalence within the region since the first reported case in 2009. This study explores the application of halophilic marine bacilli isolated from coral mucus and their quorum-quenching abilities as potential biocontrol agents in aquaculture systems to combat the causative agent of EMS, Vibrio parahaemolyticus. N-acylhomoserine lactone (AHL)-degrading (AiiA) activity was first screened by PCR then confirmed by bio-reporter assay, and a combination of 16S rDNA sequence analysis and quantitative phenotype assays including biofilm-formation and temperature-growth responses were used to demonstrate diversity amongst these quorum-quenching isolates. Three phenotypically distinct strains showing notable potential were chosen to undergo co-cultivation as a method for strain improvement via long term exposure to the pathogenic V. parahaemolyticus. The novel approach taken led to significant improvements in antagonism and quorum quenching activities as compared to the ancestral wild-type strains and offers a potential solution as well as pathway to improve existing beneficial microbes for one of the most pressing issues in shrimp aquacultures worldwide., (© 2023. The Author(s).)

For the simultaneous identification and quantification of five nitrofurans metabolites in farmed shrimp and fish, 3-amino-2-oxazolidinone (AOZ), 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ), 1-aminohydantoine (AHD), semicarbazide (SEM), and 3,5-dinitrosalicylic acid hydrazide (DNSH), an accurate, precise, and specific method was developed. The mixture of water and methanol (60/40; v/v ) was found to be the final optimised solvent for injection. The analytical run duration was 7 min, and the mobile phase included 2 mM methanol and ammonium formate. The new reference point for action (RPA) of 0.50 µg kg -1 as per EC/1871/2019 was taken into consideration and evaluated for the performance characteristics as per the CIR (EC)/2021/808 criteria. Specificity, relative retention time (≤0.25%) relative ion ratio (≤40%), linearity (0.25 to 2.0 µg kg -1 ), trueness (between 82.8 and 118.1%), repeatability (RSD r ≤14%), within lab reproducibility (RSD wr ≤16.9%), CCα (0.32-0.36 μg kg -1 ), ruggedness and relative matrix effect (≤14.26%) achieved acceptable values.

Terasi is a fermented shrimp paste in Indonesia. We examined the effect of the Terasi manufacturing process on the abundance of the allergen tropomyosin (TM) and its IgG/IgE-binding ability. Terasi was produced from three shrimps, Akiami (Acetes japonicus), Okiami (Euphausia pacifica), and Isazaami (Neomysis awatchensis). Protein degradation and TM IgE-binding activity were examined by immunoblotting using anti-TM rabbit IgG and competitive enzyme-linked immunosorbent assays using shrimp-allergic patients' sera. The processing caused TM degradation, and the IgG-specific response in Akiami meat disappeared at the second fermentation step but remained in both Okiami and Isazaami Terasi. In contrast, TM IgE-binding in all meats decreased gradually during manufacturing and nearly completely disappeared in Akiami Terasi. Conclusively, Terasi production is an effective manufacturing process to reduce the IgE-binding ability of TM, and Terasi can be recognized as a low allergenic seafood when produced under an appropriate manufacturing condition., 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 © 2022. Published by Elsevier Ltd.)

Alpha 1-acid glycoprotein (AGP) is an acute phase protein in mammals, including humans. The amount of AGP in human serum varies in response to certain diseases; thus, many efforts have been made to develop methods for quantifying human AGP. We recently discovered that luminescence occurs merely by mixing Cypridina luciferin with human AGP under human serum-free neutral or basic buffer conditions. In this study, we tested an application of Cypridina luciferin for quantifying AGP contained in human serum. Our luminescence spectrum measurements of Cypridina luciferin with human serum samples showed that the maximum emission wavelength with human serum (480 nm) differed from that with human AGP (464 nm) due to the abundant presence of endogenous human serum albumin (HSA). Furthermore, the luminescence intensities of Cypridina luciferin with human AGP in HSA-depleted human serum were consistent with those in a human serum-free basic buffer, but those in human serum were not. These results indicated that depletion of HSA in human serum was required to use Cypridina luciferin for quantifying AGP in human serum. Additionally, we found that the luminescence intensity of Cypridina luciferin with bovine AGP was approximately tenfold lower than that with human AGP., (© 2022. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.)

Body organization within arthropods is enormously diverse, but a fusion of segments into "functional groups" (tagmatization) is found in all species. Within Tetraconata/Pancrustacea, an anterior head, a locomotory thorax region, and a posterior, mostly limbless tagma known as the abdomen is present. The posterior-most tagma in crustaceans is frequently confused with the malacostracan, for example, decapod pleon often misleadingly termed abdomen, however, its evolutionary and developmental origin continues to pose a riddle, especially the completely limbless abdomen of the "entomostracan morphotype" (e.g., fairy shrimps). Since the discovery of Hox genes and their involvement in specifying the morphology or identity of segments, tagmata, or regions along the anteroposterior axis of an organism, only a few studies have focused on model organisms representing the "entomostracan morphotype" and used a variety of dedicated Hox genes and their transcription products to shine light on abdomen formation. The homeotic genes or the molecular processes that determine the identity of the entomostracan abdomen remain unknown to date. This study focuses on the "entomostracan morphotype" representative Derocheilocaris remanei (Mystacocarida). We present a complete overview of development throughout larval stages and investigate homeotic gene expression data using the antibody FP6.87 that binds specifically to epitopes of Ultrabithorax/Abdominal-A proteins. Our results suggest that the abdomen in Mystacocarida is bipartite (abdomen I + abdomen II). We suggest that the limbless abdomen is an evolutionary novelty that evolved several times independently within crustaceans and which might be the result of a progressive reduction of former thoracic segments into abdominal segments., (© 2022 The Authors. Journal of Experimental Zoology Part B: Molecular and Developmental Evolution published by Wiley Periodicals LLC.)

Early marine invertebrates like the Branchiopoda began their sojourn into dilute media some 500 million years ago in the Middle Cambrian. Others like the Mollusca, Annelida, and many crustacean taxa have followed, accompanying major marine transgressions and regressions, shifting landmasses, orogenies, and glaciations. In adapting to these events and new habitats, such invertebrates acquired novel physiological abilities that attenuate the ion loss and water gain that constitute severe challenges to life in dilute media. Among these taxon-specific adaptations, selected from the subcellular to organismal levels of organization, and constituting a feasible evolutionary blueprint for invading freshwater, are reduced body permeability and surface (S) to volume (V) ratios, lowered osmotic concentrations, increased osmotic gradients, increased surface areas of interface epithelia, relocation of membrane proteins in ion-transporting cells, and augmented transport enzyme abundance, activity, and affinity. We examine these adaptations in taxa that have penetrated into freshwater, revealing diversified modifications, a consequence of distinct body plans, morpho-physiological resources, and occupation routes. Contingent on life history and reproductive strategy, numerous patterns of osmotic regulation have emerged, including intracellular isosmotic regulation in weak hyper-regulators and well-developed anisosmotic extracellular regulation in strong hyper-regulators, likely reflecting inertial adaptations to early life in an estuarine environment. In this review, we address osmoregulation in those freshwater invertebrate lineages that have successfully invaded this biotope. Our analyses show that across 66 freshwater invertebrate species from six phyla/classes that have transmuted into freshwater from the sea, hemolymph osmolalities decrease logarithmically with increasing S:V ratios. The arthropods have the highest osmolalities, from 300 to 650 mOsmoles/kg H2O in the Decapoda with 220-320 mOsmoles/kg H2O in the Insecta; osmolalities in the Annelida range from 150 to 200 mOsmoles/kg H2O, and the Mollusca showing the lowest osmolalities at 40-120 mOsmoles/kg H2O. Overall, osmolalities reach a cut-off at ∼200 mOsmoles/kg H2O, independently of increasing S:V ratio. The ability of species with small S:V ratios to maintain large osmotic gradients is mirrored in their putatively higher Na+/K+-ATPase activities that drive ion uptake processes. Selection pressures on these morpho-physiological characteristics have led to differential osmoregulatory abilities, rendering possible the conquest of freshwater while retaining some tolerance of the ancestral medium., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

Chitin, the second richest polymer in nature, is composed of the monomer N-acetylglucosamine (GlcNAc), which has numerous functions and is widely applied in the medical, food, and chemical industries. However, due to the highly crystalline configuration and low accessibility in water of the chitin resources, such as shrimp and crab shells, the chitin is difficult utilize, and the traditional chemical method causes serious environment pollution and a waste of resources. In the present study, three genes encoding chitinolytic enzymes, including the N-acetylglucosaminidase from Ostrinia furnacalis ( OfHex1 ), endo-chitinase from Trichoderma viride ( TvChi1 ), and multifunctional chitinase from Chitinolyticbacter meiyuanensis ( CmChi1 ), were expressed in the Pichia pastoris system, and the positive transformants with multiple copies were isolated by the PTVA (post-transformational vector amplification) method, respectively. The three recombinants OfHex1, TvChi1, and CmChi1 were induced by methanol and purified by the chitin affinity adsorption method. The purified recombinants OfHex1 and TvChi1 were characterized, and they were further used together for degrading chitin from shrimp and crab shells to produce GlcNAc through liquid-assisted grinding (LAG) under a water-less condition. The substrate chitin concentration reached up to 300 g/L, and the highest yield of the product GlcNAc reached up to 61.3 g/L using the mechano-enzymatic method. A yield rate of up to 102.2 g GlcNAc per 1 g enzyme was obtained.

We characterized the existence of O-β(1,4)-GlcNAc polymers (β1,4GNP) that were anchored on the O-linked glycosylation sites of shrimp thrombospondin (pmTSP-II). There were five putative β1,4GNP linkages on the epithelial growth factor-like domain of pmTSP-II. Antibody against O-β-GlcNAc (CTD110.6) was used to prove the existence of linear and complex β1,4GNP. The antibody well reacted with linear chito-triose, -tetraose and -pentaose conjugated with phosphatidylethanolamine lipid. The immunoreactivity could also be detected with a complex β1,4GNP within pmTSP-II (at MW > 250 kDa). Upon denaturing the protein with SDS-PAGE buffer, the size of pmTSP-II was shifted to be 250 kDa, approximately 2.5 folds larger than the deduced molecular mass of pmTSP-II (110 kDa), suggesting additional association of pmTSP-II apart from its known disulfide bridging. This was confirmed by chitinase digestion on pmTSP-II protein leading to the subsequent smaller protein bands at 110-170 kDa in time- and concentration-dependent manners. These bands well reacted with CTD110.6 antibody and disappeared after extensive chitinase hydrolysis. Together, we believe that β1,4GNP on pmTSP-II serve the function in an inter-chain association to provide structural architecture of egg extracellular matrix, a novel function of pmTSP-II in reproductive biology., (© 2022. The Author(s).)

Marine crustacean waste has not been fully utilized and is a rich source of chitin. Enzymatic degradation has attracted the wide attention of researchers due to its unique biocatalytic ability to protect the environment. Chitosan (CTS) and its derivative chitosan oligosaccharides (COSs) with various biological activities can be obtained by the enzymatic degradation of chitin. Many studies have shown that chitosan and its derivatives, chitosan oligosaccharides (COSs), have beneficial properties, including lipid-lowering, anti-inflammatory and antitumor activities, and have important application value in the medical treatment field, the food industry and agriculture. In this review, we describe the classification, biochemical characteristics and catalytic mechanisms of the major degrading enzymes: chitinases, chitin deacetylases (CDAs) and chitosanases. We also introduced the technology for enzymatic design and modification and proposed the current problems and development trends of enzymatic degradation of chitin polysaccharides. The discussion on the characteristics and catalytic mechanism of chitosan-degrading enzymes will help to develop new types of hydrolases by various biotechnology methods and promote their application in chitosan.

Chitin extraction from shrimp shell powder (SSP) using protease-producing microbes is an attractive approach for valorizing shrimp shell waste because it is simple and environmentally friendly. In this study, the protease production and chitin extraction from SSP by Bacillus cereus HMRSC30 were simultaneously optimized using statistical approaches. As a result, fermentation in medium composed of 30 g/L SSP, 0.2 g/L MgSO 4 · 7H 2 O, 3 g/L (NH 4 ) 2 SO 4 , 0.5 g/L K 2 HPO 4 , and 1.5 g/L KH 2 PO 4 (pH 6.5) for 7 days maximized protease production (197.75 ± 0.33 U/mL) to approximately 1.64-fold compared to unoptimized condition (126.8 ± 0.047 U/mL). This level of enzyme production was enough to achieve 97.42 ± 0.28% deproteinization (DP) but low demineralization (DM) of 53.76 ± 0.21%. The high DM of 90% could be easily accomplished with the post-treatment using 0.4 M HCl and acetic acid. In addition, the study evaluated the possible roadmap to maximize the value of generated products and obtain additional profits from this microbial process. The observation showed the possibility of serving crude chitin as a bio-adsorbent with the highest removal capacity against Coomassie brilliant blue (97.99%), followed by methylene blue (74.42%). The recovered protease exhibited the function to remove egg yolk stain, indicating its potential for use as a detergent in de-staining. The results corroborated the benefits of microbial fermentation by B. cereus HMRSC30 as green process for comprehensive utilization of shrimp shell waste as well as minimizing waste generation along the established process., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cadmium (Cd) can adversely affect aquatic life, altering reproductive and molting processes in crustaceans. The objective of this study was to evaluate the influence of Cd on reproduction and molting in the crab Callinectes danae. Adult females were obtained from environments with different levels of pollution: low (LC), medium (MC), and high contaminated (HC) areas. Animals from LC, MC, and HC areas were exposed to 0, 0.5, and 2 mg L -1 of CdCl 2 for 3 h. Cd bioaccumulation, oxidative stress (evaluated by antioxidant enzymes activity), and lipid peroxidation (LPX) were analyzed in mature ovaries (stage II), gills, and hepatopancreas. The expression levels of crustacean hyperglycemic hormone (CHH) and molt-inhibiting hormone (MIH) genes were quantified in the eyestalks, while 17β-estradiol (E2) and melatonin concentration were measured in the hemolymph. Cd bioaccumulated mainly in the hepatopancreas and gills, with increased E2, LPX, and antioxidant enzymes in HC compared to the LC region. Decreased CHH and MIH transcripts were observed in the animals from HC regions compared to LC and MC areas. Physiological differences were recorded, especially for bioaccumulation, oxidative stress, and hormone levels, in animals sampled in HC areas compared to LC and MC regions. In conclusion, the physiological damage triggered by Cd could be reduced due to higher levels of melatonin and antioxidant enzymes in HC areas., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Manganese (Mn 2+ as MnSO 4 &/or MnCl 2 ) is a common and essential element for maintaining life in plants and animals and is found in soil, fresh waters and marine waters; however, over exposure is toxic to organisms. MnSO 4 is added to soil for agricultural purposes and people are exposed to Mn 2+ in the mining industry. Hypermanganesemia in mammals is associated with neurological issues mimicking Parkinson's disease (PD) and appears to target dopaminergic neural circuits. However, it also seems that hypermanganesemia can affect many aspects of health besides dopaminergic synapses. We examined the effect on development, behavior, survival, cardiac function, and glutamatergic synaptic transmission in the Drosophila melanogaster. In addition, we examined the effect of Mn 2+ on a sensory proprioceptive organ and nerve conduction in a marine crustacean and synaptic transmission at glutamatergic neuromuscular junctions of freshwater crayfish. A dose-response effect of higher Mn 2+ retards development, survival and cardiac function in larval Drosophila and survival in larvae and adults. MnSO 4 as well as MnCl 2 blocks stretch activated responses in primary proprioceptive neurons in a dose-response manner. Mn 2+ blocks glutamatergic synaptic transmission in Drosophila as well as crayfish via presynaptic action. This study is relevant in demonstrating the effects of Mn 2+ on various physiological functions in order to learn more about acute and long-term consequences Mn 2+ exposure., (Copyright © 2021 Elsevier Inc. All rights reserved.)

It is important to convert waste shrimp shells into chitin and its derivatives, but it still faces disadvantages of pollution and long process. In this research, deep eutectic solvents made from choline chloride (ChCl) and p-toluenesulfonic acid monohydrate (TsOH) were used to convert shrimp shells into chitin or carbon materials through adjustment of temperature with a dissolution-regeneration method in one step. Obtained precipitates and regenerated products gradually changed from chitin to carbon materials with the temperature adjusted to 130 °C and 110 °C, respectively. The α-chitin with the highest purity of 97.9 ± 0.1 wt% was prepared using 15 wt% ChCl/TsOH (1:2, n:n) aqueous solution at 70 °C, and the yield was 59.4 ± 1.9 wt%. Moreover, the carbon materials prepared by ChCl/TsOH (1:4) treatment with water and ethanol as anti-solvent were S/N/O-doped carbon materials, which exhibited potential in the supercapacitor with a specific capacitance of 24.8 and 19.3 F/g at 1 A/g, respectively., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Stomatopoda is a crustacean order including sophisticated predators called spearing and smashing mantis shrimps that are separated from the well-studied Eumalacotraca since the Devonian. The spearing mantis shrimp has developed a spiky dactyl capable of impaling fishes or crustaceans in a fraction of second. In this high velocity hunting technique, the spikes undergo an intense mechanical constraint to which their exoskeleton (or cuticle) has to be adapted. To better understand the spike cuticle internal architecture and composition, electron microscopy, X-ray microanalysis and Raman spectroscopy were used on the spikes of 7 individuals (collected in French Polynesia and Indonesia), but also on parts of the body cuticle that have less mechanical stress to bear. In the body cuticle, several specificities linked to the group were found, allowing to determine the basic structure from which the spike cuticle has evolved. Results also highlighted that the body cuticle of mantis shrimps could be a model close to the ancestral arthropod cuticle by the aspect of its biological layers (epi- and procuticle including exo- and endocuticle) as well as by the Ca-carbonate/phosphate mineral content of these layers. In contrast, the spike cuticle exhibits a deeply modified organization in four functional regions overprinted on the biological layers. Each of them has specific fibre arrangement or mineral content (fluorapatite, ACP or phosphate-rich Ca-carbonate) and is thought to assume specific mechanical roles, conferring appropriate properties on the entire spike. These results agree with an evolution of smashing mantis shrimps from primitive stabbing/spearing shrimps, and thus also allowed a better understanding of the structural modifications described in previous studies on the dactyl club of smashing mantis shrimps., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Coexistence of metals and microplastics (MPs) in aquatic environments represents a growing concern; however, little is known regarding the risks associated with their combined effects. Here, the effects of five metals (As, Cd, Cu, Pb, and Zn), alone or combined with MPs for various premixing durations (30 and 60 days), on the juvenile and adult stages of the marine mysid Neomysis awatschensis were evaluated. The toxicity (50% lethal concentration for 96 h) and bioconcentration of metals premixed with MPs were measured, and their effects on the antioxidant defense and cholinergic systems were examined. Metal toxicity increased with increasing premixing period with MPs, and juveniles were more sensitive to exposure to metals premixed with MPs than adults. Metal bioconcentration in the mysid body increased following co-exposure with MPs. Metals premixed with MPs significantly increased intracellular malondialdehyde content at both stages but decreased glutathione content in juveniles. At both stages, catalase and superoxide dismutase activity was suppressed following co-exposure to metals and MPs, except under the Cu treatment. Moreover, co-exposure inhibited acetylcholinesterase activity at both stages, suggesting cholinergic impairment. Taken together, metals and MPs produce synergistic detrimental effects on marine mysids in a stage-specific manner. Further studies are warranted to elucidate the role of MPs as a vector for contaminants and stimulator of toxicity in aquatic organisms., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Little is known about the mechanisms underlying the relationship between genetic variation and the adaptation of Oratosquilla oratoria populations to different habitat temperature. Here, the genome-wide genetic information of three O. oratoria populations were obtained by IIB restriction site-associated DNA (2b-RAD) sequencing and 2403 single-nucleotide polymorphisms (SNPs) were identified. Based on the 2403 SNPs, we found a remarkable genetic differentiation between the Yellow Sea and the East China Sea groups of O. oratoria. Furthermore, 63 SNPs are thought to be associated with different sea temperatures. Based on the 63 SNPs, it is hypothesised that the long-term temperature differences may contribute to the variation of genes associated with multiple biological functions, such as material metabolism, cytoskeleton, cellular processes, inflammatory response and hormonal regulation. This study provides new information for elucidating the molecular mechanisms underlying the relationship between genetic variation and the adaptation of Oratosquilla oratoria populations to different temperature., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Arthropods (including insects, crustaceans, and arachnids) rely on the synthesis of chitin to complete their life cycles (Merzendorfer 2011). The highly conserved chitin synthetic process and the absence of this process in vertebrates make it an exploitable target for pest management and veterinary medicines (Merzendorfer 2013; Junquera et al. 2019). Susceptible, nontarget organisms, such as insects and aquatic invertebrates, exposed to chitin synthesis inhibitors may suffer population declines, which may have a negative impact on ecosystems and associated services. Hence, it is important to properly identify, prioritize, and regulate relevant chemicals posing potential hazards to nontarget arthropods. The need for a more cost-efficient and mechanistic approach in risk assessment has been clearly evident and triggered the development of the adverse outcome pathway (AOP) framework (Ankley et al. 2010). An AOP links a molecular initiating event (MIE) through key events (KEs) to an adverse outcome. The mechanistic understanding of the underlying toxicological processes leading to a regulation-relevant adverse outcome is necessary for the utilization of new approach methodologies (NAMs) and efficient coverage of wider chemical and taxonomic domains. In the last decade, the AOP framework has gained traction and expanded within the (eco)toxicological research community. However, there exists a lack of mature invertebrate AOPs describing molting defect-associated mortality triggered by direct inhibition of relevant enzymes in the chitin biosynthetic pathway (chitin synthesis inhibitors) or interference with associated endocrine systems by environmental chemicals (endocrine disruptors). Arthropods undergo molting to grow and reproduce (Heming 2018). This process is comprised of the synthesis of a new exoskeleton, followed by the exuviation of the old exoskeleton (Reynolds 1987). The arthropod exoskeleton (cuticle) can be divided into 2 layers, the thin and nonchitinous epicuticle, which is the outermost layer of the cuticle, and the underlying chitinous procuticle. A single layer of epithelial cells is responsible for the synthesis and secretion of both cuticular layers (Neville 1975). The cuticle protects arthropods from predators and desiccation, acts as a physical barrier against pathogens, and allows for locomotion by providing support for muscular function (Vincent and Wegst 2004). Because the procuticle mainly consists of chitin microfibrils embedded in a matrix of cuticular proteins supplemented by lipids and minerals in insects (Muthukrishnan et al. 2012) and crustaceans (Cribb et al. 2009; Nagasawa 2012), chitin is a determinant factor for the appropriate composition of the cuticle and successful molting (Cohen 2001). A detailed overview of the endocrine mechanisms regulating chitin synthesis is given in Supplemental Data, Figure S1. The shedding of the old exoskeleton in insects is mediated by a sequence of distinct muscular contractions, the ecdysis motor program (EMP; Ayali 2009; Song et al. 2017a). Like the expression of chitin synthase isoform 1 (CHS-1), the expression of peptide hormones regulating the EMP is also controlled by ecdysteroids (Antoniewski et al. 1993; Gagou et al. 2002; Ayali 2009). Cuticular chitin is polymerized from uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) by the transmembrane enzyme CHS-1, which is localized in the epithelial plasma membrane in insects (Locke and Huie 1979; Binnington 1985; Merzendorfer and Zimoch 2003; Merzendorfer 2006). Because crustaceans are also dependent on the synthesis of chitin, the underlying mechanisms are believed to be similar, although less is known about different CHS isoforms and their localization (Rocha et al. 2012; Qian et al. 2014; Uddowla et al. 2014; Harðardóttir et al. 2019). Disruption of either chitin synthesis or the upstream endocrine pathways can lead to lethal molting disruption (Arakawa et al. 2008; Merzendorfer et al. 2012; Song et al. 2017a, 2017b). In the case of chitin synthesis inhibition, molting disruption can be referred to as "premature molting." If ecdysis cannot be completed because of decreased chitin synthesis, the organism may not successfully molt. Even if ecdysis can be completed on inhibition of chitin synthesis, the organism may not survive because of the poor integrity of the new cuticle. These effects are observed in arthropods following molting, which fail to survive subsequent molts (Arakawa et al. 2008; Chen et al. 2008) or animals being stuck in their exuviae (Wang et al. 2019) and ultimately dying as a result of insufficient food or oxygen intake (Camp et al. 2014; Song et al. 2017a). The term "premature molting" is used to differentiate from the term "incomplete ecdysis," which describes inhibition of ecdysis on a behavioral level, namely through reduction of the EMP (Song et al. 2017a). The present AOP describes molting-associated mortality through direct inhibition of the enzyme CHS-1. It expands the small but increasing number of invertebrate AOPs that have relevance to arthropods, the largest phylum within the animal kingdom (Bar-On et al. 2018). The development of this AOP will be useful in further research and regulatory initiatives related to assessment of CHS inhibitors and identification of critical knowledge gaps and may suggest new strategies for ecotoxicity testing efforts. Environ Toxicol Chem 2021;40:2112-2120. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC., (© 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.)

The hepatopancreas is an important digestive and immune organ in crustacean. There were low but stable numbers of microbes living in the hemolymph of crustacean, whereas the organs (including hepatopancreas) of crustacean were immersed in the hemolymph. It is very important to study the immune mechanism of the hepatopancreas against bacteria. In this study, a novel CTL (HepCL) with two CRDs, which was mainly expressed in the hepatopancreas, was identified in red swamp crayfish ( Procambarus clarkii ). HepCL binds to bacteria in vitro and could enhance bacterial clearance in vivo . Compared with the C-terminal CRD of HepCL (HepCL-C), the N-terminal CRD (HepCL-N) showed weaker bacterial binding ability in vitro and stronger bacterial clearance activity in vivo . The expression of some antimicrobial proteins, such as FLP, ALF1 and ALF5, was downregulated under knockdown of HepCL or blocked with Anti-HepCL after challenge with Vibrio in crayfish. These results demonstrated that HepCL might be involved in the antibacterial immune response by regulating the expression of antimicrobial proteins., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Cao, Pan, Sun, Liu and Lan.)

Many studies have characterized class A GPCRs in crustaceans; however, their expression in crustacean chemosensory organs has yet to be detailed. Class A GPCRs comprise several subclasses mediating diverse functions. In this study, using sequence homology, we classified all putative class A GPCRs in two chemosensory organs (antennular lateral flagellum [LF] and walking leg dactyls) and brain of four species of decapod crustaceans (Caribbean spiny lobster Panulirus argus, American lobster Homarus americanus, red-swamp crayfish Procambarus clarkii, and blue crab Callinectes sapidus). We identified 333 putative class A GPCRs- 83 from P. argus, 81 from H. americanus, 102 from P. clarkii, and 67 from C. sapidus-which belong to five distinct subclasses. The numbers of sequences for each subclass in the four decapod species are (in parentheses): opsins (19), small-molecule receptors including biogenic amine receptors (83), neuropeptide receptors (90), leucine-rich repeat-containing GPCRs (LGRs) (24), orphan receptors (117). Most class A GPCRs are predominately expressed in the brain; however, we identified multiple transcripts enriched in the LF and several in the dactyl. In total, we found 55 sequences with higher expression in the chemosensory organs relative to the brain across three decapod species. We also identified novel transcripts enriched in the LF including a metabotropic histamine receptor and numerous orphan receptors. Our work establishes expression patterns for class A GPCRs in the chemosensory organs of crustaceans, providing insight into molecular mechanisms mediating neurotransmission, neuromodulation, and possibly chemoreception., Competing Interests: The authors have declared that no competing interests exist.

Transmembrane conductance of small uncharged solutes such as glycerol typically occurs through aquaglyceroporins (Glps), which are commonly encoded by multiple genes in metazoan organisms. To date, however, little is known concerning the evolution of Glps in Crustacea or what forces might underly such apparent gene redundancy. Here, we show that Glp evolution in Crustacea is highly divergent, ranging from single copy genes in species of pedunculate barnacles, tadpole shrimps, isopods, amphipods and decapods to up to 10 copies in diplostracan water fleas although with monophyletic origins in each lineage. By contrast the evolution of Glps in Copepoda appears to be polyphyletic, with surprisingly high rates of gene duplication occurring in a genera- and species-specific manner. Based upon functional experiments on the Glps from a parasitic copepod (Lepeophtheirus salmonis), we show that such lineage-level gene duplication and splice variation is coupled with a high rate of neofunctionalization. In the case of L. salmonis, splice variation of a given gene resulted in tissue- or sex-specific expression of the channels, with each variant evolving unique sites for protein kinase C (PKC)- or protein kinase A (PKA)-regulation of intracellular membrane trafficking. The combined data sets thus reveal that mutations favouring a high fidelity control of intracellular trafficking regulation can be a selection force for the evolution and retention of multiple Glps in copepods.

Beitang shrimp paste (BSP) is fermented by different parts of shrimp, such as the head (H), meat (M), or the whole shrimp (S and W). Microbial communities of BSP were dominated by Firmicutes and Proteobacteria at the phyla level and Tetragenococcus at the genus level. However, the microbial diversity of M was the lowest than the others. Non-dominant bacterial communities were presented by a mutual symbiotic model in BSP fermentation. Tetragenococcus, Halanaerobium, Streptococcus, and Brevundimonas were positively correlated with the biosynthesis of amino acids, fatty acids, and metabolic cofactors; Marinilactibacillus and Pseudomonas might be the main contributors to inorganic sulfides, nitrogen oxides, and long-chain alkanes in BSP; Psychrobacter was closely related to the ester characteristics of methyl palmitoleate and methyl hexadecanoate in H. Halanaerobium and Streptococcus promoted the production of pyrazines in S. Tetragenococcus was positively correlated with acetic acid, decanoic acid, and palmitic acid that improved the sour aroma of M. The relationship between bacteria and aroma formation under different raw materials was expected to improve the quality of BSP., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Chitin is among the most important components of the crustacean cuticular exoskeleton and intestinal peritrophic matrix. With the progress of genomics and sequencing technology, a large number of gene sequences related to chitin metabolism have been deposited in the GenBank database in recent years. Here, we summarized the genes and pathways associated with the biosynthesis and degradation of chitins in crustaceans based on genomic analyses. We found that chitin biosynthesis genes typically occur in single or two copies, whereas chitin degradation genes are all multiple copies. Moreover, the chitinase genes are significantly expanded in most crustacean genomes. The gene structure and expression pattern of these genes are similar to those of insects, albeit with some specific characteristics. Additionally, the potential applications of the chitin metabolism genes in molting regulation and immune defense, as well as industrial chitin degradation and production, are also summarized in this review.

Shrimp are increasingly exposed to warmer temperatures and lower oxygen concentrations in their habitat due to climate change. These conditions may lead to oxidative stress and apoptosis. We studied the effects of high temperature, hypoxia, reoxygenation, and the combination of these factors on lipid peroxidation, protein carbonylation, and caspase-3 activity in gills of white shrimp Litopenaeus vannamei. Silencing of mitochondrial manganese superoxide dismutase (mMnSOD) was used to determine the role of this enzyme in response to the abiotic stressors described above, to avoid oxidative damage and apoptosis. In addition, mMnSOD gene expression and mitochondrial SOD activity were evaluated to determine the efficiency of silencing this enzyme. The results showed that there was no effect of the abiotic stress conditions on the thiobarbituric acid reactive substances (TBARS), but protein carbonylation increased in all the oxidative stress treatments and caspase-3 activity decreased in hypoxia at 28 °C. On the other hand, mMnSOD-silenced shrimp experienced higher oxidative stress, since TBARS, carbonylated proteins and caspase-3 activity increased in some silenced treatments. Unexpectedly, mitochondrial SOD activity increased in some of the silenced treatments as well. Altogether, these results suggest that mMnSOD has a key role in shrimp for the prevention of oxidative damage development and induction of apoptosis in response to hypoxia, reoxygenation, high temperature, and their interactions, as conditions derived from climate change., (Copyright © 2020 Elsevier Inc. All rights reserved.)

In vertebrates, gonadotropin-releasing hormone (GnRH) regulates gonadal maturation by stimulating the synthesis and release of pituitary gonadotropins. GnRH has also been identified in invertebrates. Crustacea consists of several classes including Cephalocarida, Remipedia, Branchiopoda (e.g., tadpole shrimp), Hexanauplia (e.g., barnacle) and Malacostraca (e.g., shrimp, crab). In the malacostracan crustaceans, the presence of GnRH has been detected in several species, mainly by immunohistochemistry. In the present study, we examined whether a GnRH-like peptide exists in the brain and/or nerve ganglion of three classes of crustaceans, the tadpole shrimp Triops longicaudatus (Branchiopoda), the barnacle Balanus crenatus (Hexanauplia), and the hermit crab Pagurus filholi (Malacostraca), by immunohistochemistry using a rabbit polyclonal antibody raised against chicken GnRH-II (GnRH2). This antibody was found to recognize the giant freshwater prawn Macrobrachium rosenbergii GnRH (MroGnRH). In the tadpole shrimp, GnRH-like-immunoreactive (ir) cell bodies were located in the circumesophageal connective of the deuterocerebrum, and GnRH-like-ir fibers were detected also in the ventral nerve cord. In the barnacle, GnRH-like-ir cell bodies and fibers were located in the supraesophageal ganglion (brain), the subesophageal ganglion, and the circumesophageal connective. In the hermit crab, GnRH-like-ir cell bodies were detected in the anterior-most part of the supraesophageal ganglion and the subesophageal ganglion. GnRH-like-ir fibers were observed also in the thoracic ganglion and the eyestalk. These results suggest that a GnRH-like peptide exists widely in crustacean species.

Heat shock proteins (HSPs) are molecular chaperones with critical roles in the maintenance of cellular proteostasis. HSPs, which regulate protein folding and refolding, assembly, translocation, and degradation, are induced in response to physiological and environmental stressors. In recent years, HSPs have been recognized for their potential role in immunity; in particular, these proteins elicit a variety of immune responses to infection and modulate inflammation. This review focuses on delineating the structural and functional roles of crustacean HSPs in the innate immune response. Members of crustacean HSPs include high molecular weight HSPs (HSP90, HSP70, and HSP60) and small molecular weight HSPs (HSP21 and HSP10). The sequences and structures of these HSPs are highly conserved across various crustacean species, indicating strong evolutionary links among this group of organisms. The expression of HSP-encoding genes across different crustacean species is significantly upregulated upon exposure to a wide range of pathogens, emphasizing the important role of HSPs in the immune response. Functional studies of crustacean HSPs, particularly HSP70s, have demonstrated their involvement in the activation of several immune pathways, including those mediating anti-bacterial resistance and combating viral infections, upon heat exposure. The immunomodulatory role of HSPs indicates their potential use as an immunostimulant to enhance shrimp health for control of disease in aquaculture., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Hemocyanins are copper-binding proteins that play a crucial role in the physiological processes in crustaceans. In this study, the cDNA encoding hemocyanin subunit 5 from the Black sea crab Eriphia verrucosa (EvHc5) was cloned using EST analysis, RT-PCR and rapid amplification of the cDNA ends (RACE) approach. The full-length cDNA of EvHc5 was 2254 bp, consisting of a 5' and 3' untranslated regions and an open reading frame of 2022 bp, encoding a protein consisting of 674 amino acid residues. The protein has an N-terminal signal peptide of 14 amino acids as is expected for proteins synthesized in hepatopancreas tubule cells and secreted into the hemolymph. The 3D model showed the presence of three functional domains and six conserved histidine residues that participate in the formation of the copper active site in Domain 2. The EvHc5 is O-glycosylated and the glycan is exposed on the surface of the subunit similar to Panulirus interruptus . The phylogenetic analysis has shown its close grouping with γ-type of hemocyanins of other crustacean species belonging to order Decapoda, infraorder Brachyura.