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Omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) have many health benefits to human. Increasing evidence have shown that climate change reduces the availability of plankton n-3 LC-PUFA to primary consumers which potentially reduces the availability of n-3 LC-PUFA to human. Since marine bivalves are an important source of n-3 LC-PUFA for human beings, and bivalve aquaculture completely depends on phytoplankton in ambient water as food, it is important to understand the impact of climate change on the lipid nutritional quality of bivalves. In this study, fatty acid profile of different bivalves (mussels, oysters, clams, scallops and cockles) from different regions (tropical, subtropical and temperate) and time (before 1990, 1991-1995, 1996-2000, 2001-2005, 2006-2010, 2011-2015, 2016-2020) were extracted from published literature to calculate various lipid nutritional quality indicators. The results of this study revealed that the effects of global warming and declines in aragonite saturation state on the lipid content and lipid indices of bivalves are highly dependent on the geographical region and bivalves. In general, global warming has the largest negative impact on the lipid content and indices of temperate bivalves, including decreasing the PUFA/SFA, EPA + DHA and n-3/n-6. However, global warming has a much smaller negative impact on lipid content and lipid indices in other regions. The declines of aragonite saturation state in seawater promotes the accumulation of lipid content in tropical and subtropical bivalves, but it compromised the PUFA/SFA, EPA + DHA and n-3/n-6 of bivalves in all regions. The findings of this study not only fill the knowledge gap of the impact of climate change on the lipid nutritional quality of bivalves, but also provide guidance for the establishment of bivalve aquaculture and fisheries management plans to mitigate the impact of climate change.

The aquaculture industry has a shortage of objected protection against soybean meal-induced enteritis (SBMIE) in carnivorous fish caused by soybean meal feed. Our initial study discovered that Lithospermum erythrorhizon (LE) ethanol extract has potential application value in improving SBMIE. A feeding trial (for eight weeks) was conducted to investigate LE ethanol extract on pearl gentian grouper SBMIE of protection to clarify the influence of LE ethanol extract on the immune tolerance profile. Three hundred and sixty pearl gentian groupers were administered one of three distinct dietary regimes: 1) 100 % fish meal (FM); 2) soybean meal substitution of 50 % fish meal protein (SBM); 3) SBM diet +0.2 % LE ethanol extract (SBMLE). Each treatment included three 1000 L cisterns-each of cisterns with 30 fish. The preliminary weight of the fish varied between 72.01 g and 72.50 g. Growth performance results showed that WGR and SGR were significantly decreased in the SBM group (P < 0.05), while there was no significant difference between the FM and SBMLE groups. There was no significant difference in survival among the three groups. The results showed that SBM-fed fish exhibited enteritis manifested by mucosal fold shortening, lamina propria widening, decreased serum immune markers (IgM, C3, and C4), and up-regulated expression of pro-inflammatory cytokines (il17 and il12) and immune-related gene (tlr3, and tlr9). The addition of 0.2 % LE ethanol extract to the SBM diet, reversed the above symptoms, and anti-inflammatory cytokine (tgf-β1), gene expression increased significantly (P < 0.05). Intestinal transcriptome analysis exhibited that the DEGs between the FM group and the SBM group were mainly enriched in FoxO signaling pathway, while the DEGs between the SBM group and the SBMLE group were enriched in MAPK signaling pathway and FoxO signaling pathway. The RT-qPCR results also revealed changes in MAPK/FoxO signaling pathway-related genes, including Dusp1, jund, Irs2b, fbxo32, and ccng2. Overall, Lithospermum erythrorhizon ethanol extract may alleviate SBMIE by regulating MAPK/FoxO signaling pathway, which would be beneficial for enhancing the immune tolerance and utilization efficiency of pearl gentian groupers to dietary soybean meal., Competing Interests: Declaration of competing interest 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 © 2024 Elsevier Inc. All rights reserved.)

Horseshoe crabs are living fossils. In recent decades, the population of horseshoe crabs, especially the tri-spine horseshoe crab Tachypleus tridentatus, has decreased significantly and was listed as an 'endangered species' under the IUCN Red List in 2019. In order to improve the reproduction of T. tridentatus to facilitate stock enhancement, it is important to understand their ovarian development. In this study, a novel TtVtg2-like gene from T. tridentatus was cloned and functionally characterized. The total legth of TtVtg2-like was 5469 bp, encoding a protein consisting of 1822 amino acid with a pI value of 6.51 and a molecular weight of 208.68 KDa. The TtVtg2-like was highly expressed in the ovary and yellow connective tissues, mainly localized in cytoplasm and endoplasmic reticulum vesicles of oocytes and yellow connective tissues, respectively. RNA interference of TtVtg2-like caused the accumulation of ROS, DNA damage, and apoptosis of ovarian primary cells. The results of this study provide useful baseline information for future studies on ovarian development in horseshoe crabs., Competing Interests: Declaration of competing interest The authors declared that there is no conflict of interest in this study. All authors have seen the manuscript and approve it to submit., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sophora flavescens is considered to be of various medicinal and economic importance. Previous studies have shown that it has antibacterial, antioxidant, and immune-enhancing functions. To explore the full potential of S. flavescens in fish feed development, we investigated the effects of different levels of S. flavescens added to the diet on the growth performance, haematological indices, and the intestinal bacterial community of Nile tilapia (Oreochromis niloticus). The feeding trial lasted for 56 days, and the initial weight of the fish was 40.00 ± 5.00 g. Nile tilapia were randomly divided into six groups: a control group (CN) and five S. flavescens supplementation groups (0.02%, 0.04%, 0.08%, 0.16%, and 0.3% [w/w] of S. flavescens in diet). The growth performance of fish increased first and then decreased with the increase of S. flavescens supplemental level. Compared with other experimental groups, the growth performance of fish supplemented with 0.08% S. flavescens was significantly improved (p < 0.05). Haematological indices exhibited that erythrocyte (RBC), along with leucocyte (WBC) indices, exhibited a secondary trend of increasing and then decreasing with the increase of S. flavescens, reaching the highest level at 0.08% (p < 0.05). However, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), and mean corpuscular haemoglobin concentration (MCHC) exhibited a secondary trend of first decreasing and then increasing with the increase of S. flavescens and reached the lowest value at 0.08% (p < 0.05). Compared with the CN, alkaline phosphatase (AKP) significantly decreased and changed when the additional amount of S. flavescens was 0.04% in each treatment group (p < 0.05). In another way, the gut microbial profiles revealed that Bacteroides dominated the gut communities, and compared with the control group, two uncultured bacteria were suppressed. In addition, when the supplemental level of S. flavescens was more significant than 0.04%, the proliferation of Arcobacter cryaerophilus, Solibacillus silvestris, and Escherichia sp. in the gut of Nile tilapia was promoted. The results revealed that S. flavescens, as an additive to the Nile tilapia diet with levels ranging from 0.04% to 0.08%, can enhance the growth performance and immunity and promote the proliferation of intestinal beneficial bacteria of Nile tilapia., (© 2024 Fisheries Society of the British Isles.)

Many researchers have evaluated the fishery carbon sink potential of bivalve aquaculture, with most studies focusing on the Life Cycle Assessment (LCA) of individual bivalves, and there is currently no consensus on whether bivalves are carbon sinks or carbon sources. It is worth noting that most studies have not considered the impact of bivalve aquaculture on ecosystems when evaluating its carbon sink potential. In this context, based on existing literature, this article aims to comprehensively review the effects of bivalve aquaculture on carbon storage in the water column and sediment of aquaculture areas. In general, our findings revealed that moderate and low stocking densities of bivalve aquaculture do not lead to significant changes in the abundance of phytoplankton, but it does indeed alter the phytoplankton community structure from dominated by huge diatom with lower carbon densities to dominated by small phytoplankton with higher carbon densities. Therefore, bivalve aquaculture may increase the total carbon storage in the water column. In addition, bivalve aquaculture also increases the sedimentation rate of suspended particles, increasing the rate of carbon burial, especially in low-energy environment and shallow water areas. The findings of this article fill the knowledge gap of fishery carbon sink in bivalve aquaculture from an ecosystem perspective., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The vitellogenin present in the bloodstream undergoes internalization into developing oocytes through the vitellogenin receptor (VgR), a process mediated by receptor-mediated endocytosis. VgR plays a crucial role in facilitating the accumulation of vitellogenin and the maturation of oocytes. In this study, we characterized a Tachypleus tridentatus vitellogenin receptor (TtVgR) gene from the tri-spine horseshoe crab, revealing a length of 1956 bp and encoding 652 amino acid residues with 12 exons. TtVgR has a molecular weight of 64.26 kDa and an isoelectric point of 5.95. Predictions indicate 85 phosphorylation sites and 7 glycosylation sites within TtVgR. Transcriptional analysis demonstrated specific expression of TtVgR in the ovary and yellow connective tissue. TtVgR was identified and distributed in the plasma membrane of oocytes. The siRNA-mediated TtVgR knockdown significantly reduced the transcriptional activity of TtVgR. This depletion induced excessive ROS production, resulting in DNA damage in ovarian primary cells. TUNEL and flow cytometry analyses confirmed ovarian cell apoptosis following TtVgR knockdown, indicating DNA damage in ovarian primary cells. These findings underscore the importance of TtVgR in ovarian cell development, suggesting its potential involvement in vitellogenesis and oocyte maturation. This knowledge may inform innovative breeding strategies and contribute to the sustainable management and conservation of the tri-spine horseshoe crab., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Dissolved oxygen (DO) is an important parameter that affects the biology, physiology, and immunology of aquatic animals. In recent decades, DO levels in the global oceans have sharply decreased, partly due to an increase in atmospheric carbon dioxide, temperature, and anthropogenic nutrient loads. Although there have been many reports on the effects of hypoxia on the survival, growth, behavior, and immunity of bivalves, this information has not been well organized. Therefore, this article provides a comprehensive review of the effects of hypoxia on bivalves. In general, hypoxia negatively impacts the food consumption rate and assimilation efficiency, as well as increasing respiration rates in many bivalves. As a result, it reduces the energy allocation for bivalve growth, shell formation, and reproduction. In severe cases, prolonged exposure to hypoxia can result in mass mortality in bivalves. Moreover, hypoxia also has adverse effects on the immunity and response of bivalves to predators, including decreased burial depths, sensitivity to predators, impairment of byssus production, and negatively impacts on the integrity, strength, and composition of bivalve shells. The tolerance of bivalves to hypoxia largely depends on size and species, with larger bivalves being more susceptible to hypoxia and intertidal species being relatively more tolerant to hypoxia. The information in this article is very useful for elucidating the current research status of hypoxia on bivalves and determining future research directions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Marine hypoxia poses a significant challenge in the contemporary marine environment. The horseshoe crab, an ancient benthic marine organism, is confronted with the potential threat of species extinction due to hypoxia, making it an ideal candidate for studying hypoxia tolerance mechanisms. In this experiment, juvenile Tachypleus tridentatus were subjected to a 21-day trial at DO:2 mg/L (hypoxia) and DO:6 mg/L conditions. The experimental timeline included a 14-day exposure phase followed by a 7-day recovery period. Sampling occurred on days 0, 7, 14, and 21, where the period from day 14 to day 21 corresponds to seven days of recuperation. Several enzymatic activities of important proteins throughout this investigation were evaluated, such as succinate dehydrogenase (SDH), phosphofructokinase (PFK), hexokinase (HK), lactate dehydrogenase (LDH), and pyruvate kinase (PK). Concurrently, the relative expression of hexokinase-1 (HK), hypoxia-inducible factor 1-alpha inhibitor (FIH), and hypoxia-inducible factor 1-alpha (HIF-1α), pyruvate dehydrogenase phosphatase (PDH), succinate dehydrogenase assembly factor 4 (SDH), and Glucose-6-phosphatase (G6Pase) were also investigated. These analyses aimed to elucidate alterations in the hypoxia signaling pathway and respiratory energy metabolism. It is revealed that juvenile T. tridentatus initiated the HIF pathway under hypoxic conditions, resulting in an upregulation of HIF-1α and FIH-1 gene expression, which in turn, influenced a shift in metabolic patterns. Particularly, the activity of glycolysis-related enzymes was promoted significantly, including PK, HK, PKF, LDH, and the related HK gene. In contrast, enzymes linked to aerobic respiration, PDH, and SDH, as well as the related PDH and SDH genes, displayed down-regulation, signifying a transition from aerobic to anaerobic metabolism. Additionally, the activity of gluconeogenesis-related enzymes such as PK and G6Pase gene expression were significantly elevated, indicating the activation of gluconeogenesis and glycogenolysis pathways. Consequently, juvenile T. tridentatus demonstrated an adaptive response to hypoxic conditions, marked by changes in respiratory energy metabolism modes and the activation of hypoxia signaling pathways., Competing Interests: Declaration of competing interest The authors state that they have no known competing financial interests or personal relationships that could affect the work reported in this article., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

PhoB is a response regulator protein that plays a key role in the PhoBR two-component signal transduction system. In this study, we used transcriptome and proteomics techniques to evaluate the detect the gene network regulated by PhoB of Streptococcus agalactiae. The results showed that expression of biofilm formation and virulence-related genes were changed after phoB deficiency. Crystal violet and CLSM assay confirmed that the deletion of the phoB increased the thickness of S. agalactiae biofilm. The results of lacZ reporter and the bacterial one-hybridization method showed that PhoB could directly bind to the promoter regions of hemolysin A and ciaR genes but not to the promoter regions of cylE and hemolysin III. Through the construction of an 18-base pair deoxyribose nucleic acid (DNA) random fragment library and the bacterial one-hybridization system, it was found that the conservative sequence of PhoB binding was TTGGAGAA(G/T). Our research has uncovered the virulence potential of the PhoBR two-component system of S. agalactiae. The findings of this study provide the theoretical foundation for in-depth research on the pathogenic mechanism of S. agalactiae., 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.)

Inadequate gonadal maturation and poor spawning performance increasingly threaten the sustainability of shrimp aquaculture. Unraveling the mechanisms regulating ovarian development and maturation hence is critical to address industry challenges. Vitellogenin (Vtg), a precursor of yolk protein found in the hepatopancreas and ovary of shrimp, plays a key role in facilitating shrimp's oocyte maturation and embryonic development after oviposition. This study found that FpVtg was specifically expressed in F. penicillatus hepatopancreas and ovary. FpVtg was localized predominantly in the oocyte cytoplasm and distributed uniformly in the hepatopancreas tissue. Silencing FpVtg led to apoptosis in both hepatopancreas and ovary tissues. Furthermore, FpVtg depletion upregulated the expression of ovarian peritrophin 1, ovarian peritrophin 2, serine proteinase inhibitor 6, and juvenile hormone esterase-like carboxylesterase 1, while downregulated that of vitellogenin, delta-9 desaturase, and insulin-like receptor. KEGG pathway analysis implicated such as PI3K-AKT signaling, RNA transport, ECM-receptor interaction, hippo signaling, oocyte meiosis, and apoptosis were enriched and involved in ovarian development. These findings have provided insights into the FpVtg's reproductive role and the associated regulatory genes and pathways in F. penicillatus. This knowledge can contribute to establishing strategies to improve the breeding and aquaculture production of F. penicillatus by elucidating its vitellogenesis regulation in redtail prawn and other penaeid species. Further characterization of the implicated pathways and genes will clarify the intricacies underlying ovarian maturation., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The insulin-like androgenic gland hormone gene (IAG), primarily expressed in the androgenic gland (AG), plays a crucial role in controlling male sex differentiation and maintaining male secondary sexual characteristics in decapods. In this study, we investigated the mRNA and microRNA expression profiles of male Procambarus clarkii to understand the transcriptomic regulatory mechanism of IAG after the injection of an efficient siRNA (GsiRNA) designed based on IAG. The results revealed that several differentially expressed genes were enriched in reproduction-related pathways, such as the wnt signaling pathway, MAPK signaling pathway, and GnRH signaling pathway. In the testis (Te), the injection of GsiRNA led to the up-regulation of many ovary-related genes and down-regulation of testis-related genes. Moreover, the brain (Br) and abdominal nerve cord (AN) appeared to be involved in the regulation of IAG, with numerous differentially expressed genes found in Br and AN. Notably, the expression of five neuropeptide genes, Crustacean hyperglycemic hormone, pigment-dispersing hormone, red pigment concentrating hormone precursor, corazonin, and gonadotropin-releasing hormone II receptor isoform X1 in Br/AN, was significantly changed. Additionally, three ovary-related miRNAs (miR-263a, miR-263b, miR-133) highly expressed in Te/AG showed significant up-regulation after GsiRNA injection. Furthermore, the long-term interference of GsiRNA was found to inhibit the development of male external sexual characteristics during the juvenile stage and delay it during the adult stage. This research provides valuable insights into the molecular regulatory mechanism and function of IAG in P. clarkii., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Vitellogenin (Vtg) serves as the precursor of yolk protein and exhibits widespread distribution in tissues, including in the ovary of both vertebrates and invertebrates. Vtg plays a critical role in facilitating oocyte maturation and embryonic development following oviposition. In this study, we have successfully elucidated the complete transcript sequence of TtVtg6-like from an ancient chelicerate Tachypleus tridentatus. The TtVtg6-like transcript encompassed a length of 4887 bp and encoded 1629 amino acids residues. Notably, TtVtg6-like was found to contain 25 exons. Furthermore, the molecular weight and isoelectric point of TtVtg6-like were determined to be 191.6 KDa and 6.73, respectively. Subsequent mRNA expression analysis demonstrated the specific expression of TtVtg6-like in ovary and yellow connective tissue. In addition, TtVtg6-like was located and distributed in both ovary and yellow connective tissue. Intriguingly, employing an siRNA approach to silence TtVtg6-like resulted in a decrease in TtVtg6-like transcription levels. Concomitantly, TtVtg6-like silencing led to increase production of ROS, ultimately resulting in DNA damage and cell apoptosis within the ovarian primary cell. The induction of apoptosis ovarian primary cells due to TtVtg6-like silencing was further corroborated through TUNEL assay and flow cytometry analysis. Overall, our findings underscore the significance of TtVtg6-like in ovarian cell development, revealing its potential association with ovarian cell apoptosis. Consequently, the insights gained from this study contribute to the future exploration of vitellogenesis and ovarian development in T. tridentatus., 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 Inc. All rights reserved.)

Radionuclide contamination in food is a public health issue. Bivalves are known to accumulate relatively high levels of radionuclides. Despite many relevant reports, this information is poorly organized. Therefore, in this study, we conducted a comprehensive scientific review of radionuclides in marine bivalves. In general, the accumulation of radionuclides in bivalves is highly species and tissue-specific, which may be due to the different biological half-life of radionuclides in different species and tissues. The trophic pathway is the main pathway for the accumulation of most radionuclides in bivalves, with polonium-210 ( 210 Po) and lead-210 ( 210 Pb) potentially selectively accumulating in the digestive glands, while 134 Cs and 137 Cs selectively accumulating in the adductor muscle and mantle. Some other radionuclides (radium-226 ( 226 Ra) and strontium-90 ( 90 Sr)) are absorbed along with other minerals (e.g. Calcium) and selectively accumulate in bivalve shells. The information in this study can provide an overview of radionuclide contamination in marine bivalves., 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.)

Bivalves have high diversity, widely distributed in various aquatic environments, including saltwater, brackish water and freshwater. Bivalves are known to rich in polysaccharides and have wide applications in functional foods, pharmaceuticals, and industrial research. Despite many relevant reports are available, the information is poorly organized. Therefore, in this study, we conducted a comprehensive scientific review on the potential bioactivity of polysaccharides derived from bivalves. In general, the polysaccharides derived from bivalves possess various bioactive properties, including anticancer, antioxidant, anticoagulant and immunomodulatory activities. The bioactivity of these biomolecules highly depends on the bivalve species, extraction methods, purification methods, dosages, etc. The information in this study can provide an overview of the bioactivities of bivalve polysaccharides. This is very useful to be used as a guide for identifying the health benefits of polysaccharides derived from different bivalve species., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

CD46, as a cofactor of complement I factor, not only regulates the complement system but also functions as a pathogen receptor and is involved in controlling early pathogen infection through autophagy. In this study, a new CD46 gene (ToCD46) was identified from golden pompano (Trachinotus ovatus), which showed higher sequence homology with other teleosts CD46. Homology comparison showed that ToCD46 had higher sequence homology (46.95-52.85%) with other teleosts CD46 and lower homology with mammal. Tissue expression profile analysis showed that ToCD46 was generally expressed in all tissues with the highest expression level in liver, followed by head kidney, and showed different patterns of up-regulation in immune-related tissues after stimulation by Streptococcus agalactiae and Vibrio alginolyticus. The hemolytic activity analysis and apoptosis assay showed that rToCD46 decreased the hemolytic activity of serum of golden pompano and effectively inhibited the damage of A549 cells, suggesting that ToCD46 might be involved in the regulation of complement activation of golden pompano. In vitro antibacterial experiments showed that rToCD46 had antibacterial activity against gram negative bacteria V. alginolyticus but no effect on positive bacteria S. agalactiae. These results suggest that ToCD46 may be involved in the immune response of golden pompano to pathogens, which will provide important basic information for elucidating the evolutionary history of the complement system of golden pompano., 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.)

Coronary heart disease (CHD) is one of the leading causes of death worldwide. Seafood, especially fish and shellfish, is a healthy food that reduces the risk of CHD. In many regions, seafood is consumed cooked to eliminate potentially pathogenic microorganisms. Although there have been many reports of culinary preparations causing changes in the fatty acid profile of fish and shellfish, this information has not been well organized, and most of it is not associated to CHD. Therefore, this study was conducted to study the effect of culinary treatments of seafood on lipid nutritional quality in relation to promotion/prevention of CHD. In this study, fatty acid profiles of fish and shellfish prepared with different culinary preparations were obtained from published literature. Lipid nutritional quality indices related to promoting/preventing CHD were calculated and analyzed to reveal the effects of culinary treatment on the lipid nutritional quality of fish and shellfish in promoting/preventing of CHD. The information in this article is very useful and can fill the knowledge gap of the effects of culinary preparation on the lipid nutritional quality of fish and shellfish. Such information is very useful for guiding consumers to choose better ways to cook fish and shellfish to reduce the risk of CHD., Competing Interests: 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., (© 2023 The Author(s).)

Backgroud: Macrobrachium rosenbergii is an economically important species that is widely cultivated in some Asian nations. Foxl2 is a transcriptional regulator of ovarian differentiation and development. The aim of this study was to study the bioinformatics features and expression patterns of M. rosenbergii Foxl2 (MrFoxl2)., Methods: In this study, all experimental animals were mature M. rosenbergii (9-12 cm) individuals. The foxl2 gene was identified and characterized in the genome of M. rosenbergii using molecular cloning, bioinformatic analysis, in situ hybridization, and quantitative analysis., Results: The identified cDNA encoded a putative 489-amino-acid MrFoxl2 protein. Bioinformatics analysis revealed a low identity of MrFoxl2 to other crustacean orthologues. The closest phylogenetic relationship was to Foxl2 of Eriocheir sinensis. The result of in situ hybridization demonstrated that transcripts of MrFoxl2 in M. rosenbergii were identified in spermatocytes, oocytes, and secretory epithelial cells of the vas deferens. The result of q-PCR suggested that a high expression of MrFoxl2 was identified in the testis, vas deferens, and ovaries. During ovarian development, MrFoxl2 expression was the highest in stage I., Conclusion: Our findings suggest that MrFoxl2 may play a role in gonadal development in both female and male M. rosenbergii., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Pancreatic ribonuclease (RNase1) of Megalobrama amblycephala exhibits both antimicrobial and digestive activity. The gut microbiome improve the digestion and metabolic capacity and enhance the functioning of the immune system of the host against pathogenic bacteria. In this study, we aimed to assess the protective effect of RNase1 on Aeromonas hydrophila-induced inflammation and intestinal microbial metabolism. Megalobrama amblycephala were randomly divided into three groups: control (injected PBS), infection (A. hydrophila-injected), and treatment group (RNase1 pretreatment 24 h before the A. hydrophila injection). The morphological symptoms were significantly alleviated by RNase1. RNase1 reshaped the perturbed gut microbiota by upregulating Proteobacteria and Vibrio richness and downregulating Firmicutes, Chlamydiae, Bacillus, and Gemmobacter richness. The lysophosphatidylcholine, (±) 17 HETE, D- (+) -cellobiose, and PC (20:5) in the treatment group were restored by RNase 1 protein treatment to the level of the control group. In the treatment group, phospholipid metabolism, fatty acid metabolism, glucose metabolism and lipid metabolism were different from the control and infection groups. The proinflammatory factors concentration in intestinal samples significantly increased after A. hydrophila infection. Our results revealed that RNase1 plays an important role in resistance to pathogen invasion, reducing inflammation, and improving intestinal function, thus inhibiting the occurrence of disease., (© 2021 Society for Applied Microbiology and John Wiley & Sons Ltd.)

RNA interference (RNAi), which employs double-strand RNA (dsRNA) or small interference RNA (siRNA), is a popular reverse genetic manipulation tool to study gene function. Presently, there is few reports on the implementation of RNAi on the insulin-like androgenic gland gene (IAG) in red swamp crayfish Procambarus clarkii. In this study, the effective sequence of siRNA and optimal injection dose were determined, and the effects of RNAi using dsRNA, siRNA, and long-term RNAi were investigated. The results showed that the doses of 0.5 and 1 µg/g of body weight of IAG-siRNA3 produced significantly better inhibition than 0.1 µg/g. qPCR assays showed that both dsRNA and siRNA silenced the IAG expression in five tissues (brain, ventral nerve cord, androgenic gland, testis, and vas deferens) in adult P. clarkii, with the effectiveness decreasing over time, inhibiting the production of spermatid. dsRNA exhibited a longer interference effect than siRNA in adults. For long-term interference (P. clarkii juveniles were injected 7 times with 1 µg/g of body weight of IAG-dsRNA), and found that the secondary sexual characteristics of juveniles were affected, while the control group developed normally. The results of this study could lay the foundation for crayfish sex reversal with IAG RNAi, and provide the reference for those studies in which the technique of RNAi was used., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The insulin-like androgenic gland (IAG) gene is well known in male crustacean, and it is a key regulator in male sexual differentiation and maintaining the male sexual characteristic. The neo-female can be produced by silencing the MrIAG (Macrobrachium rosenbergii Insulin-like Androgenic Gland) in male Macrobrachium rosenbergii. This is the first time to use siRNA approach to silenced MrIAG in male M. rosenbergii. In the current study, the optimal injection dosage to achieve sex reversal is 0.5 μg/g body weight. After MrIAG silencing, the expression level of Dmrt11e, Dmrt99b, MRPINK, Mrr, Sxl1, and Sxl2 decreased significantly. As their long-term silencing effect of MrIAG, the dsRNA and siRNA approaches produce three and two individual neo-females, respectively. The neo-female has a wider brood chamber, ovipositing setae, and ovigerous setae, which is resembled normal female. After a long-term silencing with siRNA, most of the germ cells were arrested in spermatocytes stage, but the spermatocytes in control can further developed into spermatozoon. The seminiferous tubules are loosely arranged and the spermatocytes are more than spermatozoon in the 0.5 μg/g body weight treatment dose. This current study suggests a new path to obtain neo-females through siRNA silencing.

Toll-like receptors (TLRs) are a category of pattern recognition receptors (PRRs), which recognize pathogen associated molecular patterns (PAMPs) and participate in the immune responses. We identified tlr5a, tlr5b, tlr9 and tlr21 from the genome of blunt snout bream (Megalobrama amblycephala). All four tlrs were constitutively expressed in all examined tissues. After an immune bacterial challenge with Aeromonas hydrophila, their expressionwas up-regulated in lymphoid organs and tissues. Recombinant eukaryotic plasmid pEGFP-N1 was transfected into the common carp (Cyprinus carpio) EPC (epithelioma papulosum cyprini) cells for the purpose of subcellular localization. pcDNA3.1(+) recombinant eukaryotic plasmid was used to investigate the effects of overexpression of tlrs on the expression of downstream interferon-associated immune factors. The four Tlrs were distributed in the cytoplasm of transfected cells and appeared as filamentous or reticular. The expression of irf3, irf7, isg15, mx1, pkr and viperin at 0, 6, 12, 18, 24, 36, 48 and 72 h post-transfection in transfected EPC cells was quantified by qPCR. Overexpression of tlrs upregulated the expression of viperin, isg15, irf3, irf7, mx1 and pkr (in that order of magnitude). We also cloned the following promoters of irfs: Irf1-p, irf2-p, irf6-p, irf7-p, irf8-p and irf9-p. Results of the dual luciferase reporter assay suggested that tlr5a, tlr5b and tlr9 enhanced the activities of irf7-p, while tlr5b enhanced the activities of irf1-p and irf7-p. This suggests that they all play a role in the innate immunity. The experiments also indicated that TLRs activate irf3 or irf7 signaling to induce IFN secretion and subsequent upregulation of IFN-stimulated genes. These results indicate that tlrs and irfs play an important immune role in response to A. hydrophila infection in blunt snout bream, and pave the way for further studies of immune mechanisms mediated by TLRs in fish., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Paramisgurnus dabryanus is one of the most economically important fishes in China. Barbels are an essential sensory organ for the food-seeking ability of teleost fish. However, the anatomical structure of the maxillary barbels of P. dabryanus and the molecular basis of their development are unknown. We investigated the anatomical structure of the barbel, and gene expression patterns of two chemokine C-C motif ligands: CCL4 and CCL13-like during the maxillary barbel development using Masson Trichrome staining, light and electron microscopy, and qPCR. Anatomically, the maxillary barbel of P. dabryanus contains taste buds, melanophores, collagen fibers, connective tissue, smooth muscles, nerve bundles, and blood vessels, but does not have skeletal muscles or a skeleton rod. The expression of CCL4 and CCL13-like was weak or non-existent in the early phases of development, but high at the last two studied time-points: 192- and 216-h post-hatching. Results indicated that CCL4 and CCL13-like were related to the development of the maxillary barbel.