Aquatic Organisms Research Articles

Comparison of the Intestinal Microbiota Composition and Function of Red Claw Crayfish (Cherax quadricarinatus) Cultured in Ponds and Rice Fields

The growth environment significantly influences the intestinal microbiota of aquatic organisms. We investigated the composition and functional differences in the intestinal microbiota of red claw crayfish (Cherax quadricarinatus) in rice fields (RB) and ponds (PB) by 16S rDNA high-throughput sequencing technology. The results indicate that the Shannon, Simpson, Sobs, Chao1, and ACE indices of PB are all higher than those of RB, demonstrating greater diversity and richness of intestinal microbiota. The dominant phyla in the intestinal microbiota of the Cherax quadricarinatus were Proteobacteria, Tenericutes, and Firmicutes. Tenericutes and Proteobacteria were significantly more abundant in the RB than in the PB, while Planctomycetes and Firmicutes were significantly more abundant in the PB than in the RB. The results of network correlation analysis indicate that Proteobacteria and Firmicutes exhibit strong connectivity with other microbial groups in the gut microbiota of Cherax quadricarinatus, showing significant centrality. They play an important role in the interactions within the gut microbiota community. The dominant bacterial genera in the Cherax quadricarinatus’s gut were Citrobacter, Candidatus_Bacilloplasma, and Clostridium_sensu_stricto_1. The abundance of the genus Clostridium was significantly higher in the PB than in the RB, whereas the abundance of Candidatus_Hepatoplasma and Vibrio was significantly lower in the PB than in the RB. The prediction function of KEGG enrichment showed that the abundance of Amino acid metabolism, Biosynthesis of Other Secondary Metabolites, Transport and Catabolism, Cancers, and Nervous System, Substance Dependence were significantly higher in the PB, while the infectious diseases pathway was enriched in the RB. In summary, our results revealed significant differences in the composition and diversity of intestinal microbiota in the Cherax quadricarinatus between rice paddy and pond farming environments. The intestinal microbiota of the Cherax quadricarinatus grown in pond environments exhibit higher diversity and stability, manifested by an increase in beneficial bacteria abundance and a decrease in opportunistic pathogens. These findings significantly improve understanding of the complex relationship among Cherax quadricarinatus, intestinal microbiota, and the environment.

Visible Combined Near-Infrared in Situ Imaging Revealed Dynamic Effects of Microplastic Fibers and Beads in Zebrafish.

Microplastics (MPs) as emerging contaminants are widely present in the environment and are ubiquitously ingested and accumulated by aquatic organisms. MPs may be quickly eliminated after a brief retention in aquatic animals (such as the digestive tract); thus, understanding the damage caused by MPs during this process and whether the damage can be recovered is important. Here, we proposed the use of visible light imaging to track MPs combined with near-infrared (NIR) imaging to reveal the in situ impacts of MPs. The combination of these two techniques allows for the simultaneous investigation of the localization and functionality of MPs in vivo. We investigated the effects of two types of MPs on zebrafish, microplastic fibers (MFs) and microplastic beads (MBs). The results showed that MPs larger than 10 μm primarily accumulated in the intestines of zebrafish. Both MFs and MBs disrupted the redox balance of the intestine, and the location of the damage was consistent with the heterogeneous accumulation of MPs. MFs caused greater and more difficult-to-recover damage compared to MBs, which was closely related to the slower elimination rate of MFs. Our study highlights the importance of capturing the dynamic toxicological effects of MPs on organisms. Fibrous MPs and spherical MPs clearly had distinct effects on their toxicokinetics and toxicodynamics in fish.

Microplastics are effective carriers of bisphenol A and facilitate its escape from wastewater treatment systems.

Microplastics (MPs) pollution is a major issue in aquatic environments. Wastewater treatment plants are significant point sources of MPs, which may also be carriers of organic pollutants. We analyzed MP number, shape, color, and polymer type distribution in sewage wastewater treatment plants. The potential of MPs to act as carriers for typical organic pollutants in sewage, such as bisphenol A (BPA), was also assessed. The predominant MPs in the influent were fibers, primarily transparent and black in color, and composed of polyethylene, polypropylene, and polystyrene. During wastewater treatment, the concentration of MPs decreased from 10.89 items per L in the influent to 0.89 items per L in the treated effluent, with significant differences in treatment efficiency at different stages. In the simulated wastewater, the three predominant MPs exhibited certain adsorption capacities for bisphenol A. Changing the temperature and pH within the range expected for wastewater could interfere with the interactions between MPs and bisphenol A, with a limited impact on adsorption. The results show that although wastewater treatment plants intercept a significant amount of MP, a considerable number of them enter the aquatic environment daily because of the high volume of wastewater discharge. These MPs, which carry pollutants such as bisphenol A, may threaten the health of fish and other aquatic organisms. However, by scientifically adjusting operational parameters, wastewater treatment plants could become "controllable sources" of MP compound pollutants.

Assessing habitat selection in the prawn Macrobrachium rosenbergii using the model toxicant copper and colonization as a test endpoint: Does prior exposure determine biochemical and behavioral responses?

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

Mechanisms insights into bisphenol S-induced oxidative stress, lipid metabolism disruption, and autophagy dysfunction in freshwater crayfish

Bisphenol S (BPS) is widely used in plastic products, food packaging, electronic products, and other applications. In recent years, BPS emissions have increasingly impacted aquatic ecosystems. The effects of BPS exposure on aquatic animal health have been documented; however, our understanding of its toxicology remains limited. This study aimed to explore the mechanisms of lipid metabolism disorders, oxidative stress, and autophagy dysfunction induced in freshwater crayfish (Procambarus clarkii) by exposure to different concentrations of BPS (0 µg/L, 1 µg/L, 10 µg/L, and 100 µg/L) over 14 d. The results indicated that BPS exposure led to oxidative stress by inducing elevated levels of reactive oxygen species (ROS) and inhibiting the activity of antioxidant-related enzymes. Additionally, BPS exposure led to increased lipid content in the serum and hepatopancreas, which was associated with elevated lipid-related enzyme activity and increased expression of related genes. Furthermore, BPS exposure decreased levels of phosphatidylcholine (PC) and phosphatidylinositol (PI), disrupted glycerophospholipid (GPI) metabolism, and caused lipid deposition in the hepatopancreatic. These phenomena may have occurred because BPS exposure reduced the transport of fatty acids and led to hepatopancreatic lipid deposition by inhibiting the transport and synthesis of PC and PI in the hepatopancreas, thereby inhibiting the PI3K-AMPK pathway. In conclusion, BPS exposure induced oxidative stress, promoted lipid accumulation, and led to autophagy dysfunction in the hepatopancreas of freshwater crayfish. Collectively, our findings provide the first evidence that environmentally relevant levels of BPS exposure can induce hepatopancreatic lipid deposition through multiple pathways, raising concerns about the potential population-level harm of BPS and other bisphenol analogues.

The toxic effect of 2,6-di-tert-butylphenol on embryonic development in zebrafish (Danio rerio): Decreased survival rate, morphological abnormality, and abnormal vascular development

2,6-di-tert-butylphenol (2,6-DTBP) has been used extensively in plastics, rubber and polymer phenolic antioxidants. It is discharged into the aquatic environment through industrial waste. However, the toxicity assessment of 2,6-DTBP is insufficient. Here, zebrafish embryos were used as an animal model to investigate the toxicological effects of 2,6-DTBP. The results showed that 2,6-DTBP induced mitochondrial dysfunction and reactive oxygen species accumulation, which caused apoptosis, and further led to developmental toxicity of zebrafish embryos, such as delayed incubation, reduced survival rate, and increased malformation rate and heart rate. 2,6-DTBP can also cause morphological changes in the zebrafish endothelial cell (zEC) nucleus, inhibit zEC migration, trigger abnormal angiogenesis and zEC sprouting angiogenesis, and ultimately affect vascular development. In addition, 2,6-DTBP interfered with the endogenous antioxidant system, causing changes in activities of superoxide dismutase, catalase, and glutathione S-transferase and contents of malondialdehyde and glutathione. Transcriptome sequencing showed that 2,6-DTBP altered the mRNA levels of genes associated with vascular development, oxidative stress, apoptosis, extracellular matrix components and receptors. Integrative biomarker response assessment found that 12 μM 2,6-DTBP had the highest toxicity. These results indicated that 2,6-DTBP induced apoptosis through oxidative stress, leading to toxicity of zebrafish embryo development. This study contributes to understanding the effects of environmental 2,6-DTBP exposure on early development of aquatic organisms and draws public attention to the health risks posed by chemicals in aquatic organisms.

Host-associated microbes mitigate the negative impacts of aquatic pollution.

Pollution can negatively impact aquatic ecosystems, aquaculture operations, and recreational water quality. Many aquatic microbes can sequester or degrade pollutants and have been utilized for bioremediation. While planktonic and benthic microbes are well-studied, host-associated microbes likely play an important role in mitigating the negative impacts of aquatic pollution and represent an unrealized source of microbial potential. For example, aquatic organisms that thrive in highly polluted environments or concentrate pollutants may have microbiomes adapted to these selective pressures. Understanding microbe-pollutant interactions in sensitive and valuable species could help protect human well-being and improve ecosystem resilience. Investigating these interactions using appropriate experimental systems and overcoming methodological challenges will present novel opportunities to protect and improve aquatic systems. In this perspective, we review examples of how microbes could mitigate negative impacts of aquatic pollution, outline target study systems, discuss challenges of advancing this field, and outline implications in the face of global changes.

Histomorphological and Dynamical Changes in Female River Lampreys during Maturation under Controlled Conditions as a Part of Lamprey Restoration Programs.

More than 40 species of lampreys (Petromyzontiformes) are known worldwide. Some of them are parasitic and feed on the blood of fish or other aquatic animals. Lampreys spawn once in their lifetime, after which they die. One of the representatives of the ichthyofauna of European rivers is the river lamprey, Lampetra fluviatilis. The river lamprey is now an endangered species due to loss and degradation of their habitats. The present study investigated gonadal development without hormonal stimulation in female river lampreys during puberty under controlled conditions for a period of seven months. Female river lampreys were kept in conditions that mimicked the natural environment. During the November-May period, samples were taken monthly to determine the extent of gonadal development and gastrointestinal and liver changes using histological examination. From the results obtained, the dynamical changes were determined for the following: gonadosomatic index (GSI; %), hepatosomatic index (his; %), and digestivesomatic index (DSI; %). With the gonadal development of female lampreys, an increase in GSI (7-23%; November-May) and a decrease in DSI (0.4-0.1%; November-May) histological changes were observed in the gonads (oocyte development), intestines (over time, decreased lipid vacuoles and enterocyte apoptosis), and in the liver (decreased lipid vacuoles and hepatocyte apoptosis over time) and in the digestive system resulting from its degradation. Also, it was observed that there was a change in the color of the liver to green in April. This study demonstrated the key role of the liver in the oocyte maturation process in this species.

Application of terpenoids for the remediation of environmental water polluted with bisphenol A and its analogs using an in silico approach

Nowadays, there is a global concern over water quality and the impact of contamination on both natural ecosystems and human well-being. Plastics, ubiquitous in modern life, may release harmful chemicals when they reach aquatic environments. Among them, bisphenol A (BPA) and its alternatives, such as bisphenol S (BPS), bisphenol F (BPF), and others, are of special concern because their presence in water systems can have detrimental effects on human health and aquatic organisms due to their endocrine-disrupting properties.This study explores the potential of terpenoids, sustainable and environmentally friendly solvents, for efficiently removing bisphenols from contaminated environmental water. Using an in silico approach based on the Conductor-like Screening Model for Realistic Solvents (COSMO-RS) theory, >30 terpenoids were screened, and carvone was found to be an excellent candidate due to its high solvent capacity and low toxicity. The impact of pH, temperature, stirring conditions, and sample:extractant phase ratios on the extraction efficiency were investigated. A design of experiments revealed optimal conditions for the extraction process and demonstrated that carvone can effectively extract bisphenols (nearly 100 % for most of them) under a wide range of conditions, showing the robustness and efficiency of the extraction method, even in environmental samples.The paper provides valuable insights into the potential of terpenoids, specifically carvone, as a sustainable and eco-friendly solvent for removing bisphenol contaminants from environmental water bodies. The findings of this study offer a promising solution to address water contamination issues, aligning with the principles of Green Chemistry and contributing to a more environmentally responsible approach to water remediation.

A water strider-inspired intestinal stent actuator for controllable adhesion and unidirectional biofluid picking

Soft-bodied aquatic organisms exhibit extraordinary navigation and mobility in liquid environments which inspiring the development of biomimetic actuators with complex movements. Stimulus-responsive soft materials including hydrogels and shape-memory polymers are replacing traditional rigid parts that leading to dynamic and responsive soft actuators. In this study, we took inspiration from water strider to develop a biomimetic actuator for targeted stimulation and pH sensing in the gastrointestinal tract. We designed a soft and water-based Janus adhesive hydrogel patch that attaches to specific parts of the intestine and responds to pH changes through external stimulation. The hydrogel patch that forms the belly of the water strider driver incorporates an inverse opal microstructure that enables pH responsive behavior. The hydrogel patch on the water strider's leg uses a sandwich structure of Cu particles to convert light into heat and bend under infrared light to mimic the water strider's leg simulating the efficient and steady movement of the water strider's leg which transporting the biological fluid in one direction. This miniature bionic actuator demonstrates controlled adhesion and unidirectional biofluid delivery capabilities, proving its potential for targeted stimulus response and pH sensing in the gastrointestinal tract, thus opening up new possibilities for medical applications in the growing field of soft actuators.

The Australian Biosecurity Genomic Database: a new resource for high-throughput sequencing analysis based on the National Notifiable Disease List of Terrestrial Animals.

The Australian Biosecurity Genomic Database (ABGD) is a curated collection of reference viral genome sequences based on the Australian National Notifiable Disease List of Terrestrial Animals. It was created to facilitate the screening of high-throughput sequencing (HTS) data for the potential presence of viruses associated with notifiable disease. The database includes a single verified sequence (the exemplar species sequence, where relevant) for each of the 60 virus species across 21 viral families that are associated with or cause these notifiable diseases, as recognized by the World Organisation for Animal Health. The open-source ABGD on GitHub provides usage guidance documents and is intended to support building a culture in Australian HTS communities that promotes the use of quality-assured, standardized, and verified databases for Australia's national biosecurity interests. Future expansion of the database will include the addition of more strains or subtypes for highly variable viruses, viruses causing diseases of aquatic animals, and genomes of other types of pathogens associated with notifiable diseases, such as bacteria. Database URL: https://github.com/ausbiopathgenDB/AustralianBiosecurityGenomicDatabase.

Evaluation on the antibacterial activity of glycyrrhizin against Pseudomonas plecoglossicida in ayu fish (Plecoglossus altivelis)

Glycyrrhizin (GLR), an active constituent of the traditional Chinese medicine (TCM) liquorice (Glycyrrhiza uralensis Fisch), has important functions against bacterial invasions in mammals. However, studies on the antimicrobial properties of GLR in teleosts remain relatively scarce. Herein, we performed an in vitro screening of 30 monomeric TCM compounds and found that only GLR exhibited significant inhibitory activity against the bacterium Pseudomonas plecoglossicida and increased the survival rate of ayu fish challenged with it. Moreover, we explored the antimicrobial properties of GLR in in vitro antimicrobial experiments and found that it inhibited all 10 common aquatic pathogens, including P. plecoglossicida. Notably, spectrophotometry and fluorescence staining assays revealed that GLR exhibited bactericidal activity in vitro by inhibiting P. plecoglossicida biofilm formation and increasing its membrane permeability. Notably, we showed that GLR could reduce pro-inflammatory factor levels, reduce bacterium-triggered apoptosis of monocytes/macrophages (MO/Mф), and promote their phagocytosis and bactericidal capacity. Mechanically, a cellular thermal shift assay confirmed that GLR could bind to high mobility group box 1 (HMGB1), suggesting that GLR may regulate MO/Mф functions through HMGB1-mediated pathways. Overall, our findings confirm the direct bactericidal ability of GLR and resistance to pathogen invasion by modulating the immune activity of MO/Mф, providing an essential basis for the utilisation of TCM in aquatic animal protection.

The novel peptide PEP-Z-2 potentially treats renal fibrosis in vivo and in vitro by regulating TGF-β/Smad/AKT/MAPK signaling

Renal fibrosis is a process in which excessive deposition of extracellular matrix leads to an increase in tissue hardness and gradual destruction of the renal parenchyma. Chronic kidney disease (CKD) commonly progresses to end-stage renal disease (ESRD), ultimately leading to renal failure. This disease has high incidence and mortality rates, but to date, effective treatment options are lacking. PEP-Z-2 is a collagen peptide isolated from redlip croaker scales and may have potential fibroprotective activity. In this study, PEP-Z-2 was found to alleviate unilateral ureteral obstruction (UUO)- and folic acid (FA)-induced kidney injury in a mouse model, reduce collagen deposition in tissues, normalize renal function, reduce the expression of fibrosis markers, reduce reactive oxygen species (ROS) production, and restore the balance of the oxidant/antioxidant system. In vitro experiments also demonstrated that PEP-Z-2 inhibits the TGF-β-induced differentiation of fibroblasts and renal tubular epithelial cells into myofibroblasts and reduces the production of extracellular matrix (ECM) proteins such as fibronectin, Col I, and α-SMA, demonstrating notable therapeutic effects on renal fibrosis. This effect is achieved by regulating the TGF-β/Smad/AKT/MAPK pathway. Our research suggested that PEP-Z-2 is a potential therapeutic drug for renal fibrosis, and peptides from aquatic organisms may constitute a new class of candidate drugs for the treatment of renal fibrosis and even other types of organ fibrosis.

Zinc oxide nanoparticles cause hepatotoxicity in rare minnow (Gobiocypris rarus) via ROS-mediated oxidative stress and apoptosis activation and inhibition of lipid peroxidation

Zinc oxide nanoparticles (ZnO NPs) measuring 30 ± 10 nm in diameter are utilized in various applications, including batteries, plastics, ceramics, cosmetics, flame retardants, and food. However, their potential impact and risk on aquatic ecosystems remain unclear. This study examined the hepatotoxic effects of dietary ZnO NPs in rare minnow. Three hundred rare minnows were fed diets containing 0 mg/kg, 20 mg/kg and 60 mg/kg of ZnO NPs for 60 days, with hepatotoxicity assessments at 15-day intervals. The histological data showed that ZnO NPs severely damage liver tissues, causing cytoplasmic vacuolization and irregular or missing nuclei. The treatment groups showed a significant rise in the liver injury index (P < 0.05). Moreover, there was a notable increase in zinc accumulation in the ZnO NPs groups compared to the control group (P < 0.05). ZnO NPs also reduced body weight and hepato-somatic index (HSI) in rare minnows. The enzyme activity results showed elevated levels of reactive oxygen species (ROS), total cholesterol (T-CHO), triglyceride (TG), acetyl Coa carboxylase (ACC), and fatty acid synthase (FAS) in the ZnO NPs fed groups, while total antioxidant capacity (T-AOC) and malondialdehyde (MDA) decreased. Further investigation found that accumulation of ZnO NPs in the liver tissues up-regulated the levels of genes related to antioxidant (mn-sod, cat and nf-κb), pro-apoptotic (bax) and lipogenesis (gpat3, dgat1a and dgat1b), while down-regulating genes associated with lipid catabolism (cpt1 and tpi1). These findings suggest that dietary ZnO NPs cause hepatotoxicity by inducing oxidative stress through ROS, triggering apoptosis, and inhibiting lipid peroxidation. The results indicate that ZnO NPs may pose a significant threat to aquatic animals and ecosystems.

Ecotoxicity of the chlorantraniliprole insecticide on Ceriodaphnia dubia

Studies on the toxic effects of pesticides to aquatic organisms are necessary to establish safe concentrations of exposure, promoting the protection of aquatic life. The insecticide chlorantraniliprole has been used since 2007 in several crops of vegetables and fruits, but despite its wide and continuous use, little ecotoxicological information is reported in the literature. Then, the objective of this work is to evaluate the ecotoxicological aspects of this insecticide in the test organism Ceriodaphnia dubia through toxicity tests, aiming at determining the values of NOEC (no observed effect concentration), LOEC values (lowest observed effect concentration), CV (chronic value) and EC(I)50, 48 h (acute effect concentration). The results indicated that C. dubia showed greater sensitivity when compared to other organismal groups exposed to the same insecticide, with EC(I)50, 48 h values of 2.9 ± 1.11 µg.L-1, NOEC of 0.61 ± 0.33 µg.L-1 and LOEC of 0.91 ± 0.49 µg.L-1. Based on toxicity tests with C. dubia and considering the protection of aquatic life, a safe environmental concentration of 0.74 µg. L-1 of the insecticide chlorantraniliprole is recommended.

Encapsulation and release of tetrabromobisphenol A in microplastics: Trade-off in their individual toxicity to Xenopus tropicalis tadpoles

The toxicity of microplastics (MPs) to aquatic animals is closely related to the presence and release kinetics of contained additives, as most plastic products contain various additives. However, the relationship between the occurrence and release of additives from MPs, and their individual or combined toxicity remains unclear. In this study, the nanoscale distribution and release of tetrabromobisphenol A (TBBPA, a common flame retardant with endocrine-disrupting effect) in polystyrene (PS) MPs, and the long-term (60 days) toxicity of TBBPA and MPs containing TBBPA (at doses of 0 %, 1 %, 10 %, w/w) to Xenopus tropicalis tadpoles were investigated. Exposure to 10 μg/L TBBPA alone was the most toxics, while the encapsulation of TBBPA in MPs significantly delayed its lethal toxicity to tadpoles by inhibiting the rapid and extensive release of TBBPA. PS MPs alone and MPs containing 10 % TBBPA exhibited delayed survival toxicity compared to TBBPA alone, whereas PS MPs containing 1 % TBBPA did not show this effect but inhibited growth. These findings suggest that chronic toxicity assessments should be based on long-term (months or even years) exposure experiments due to the encapsulation-controlled slow release of toxic additives.

Effects of Dietary Bio-Fermented Selenium Supplementation on Growth, Immune Performance, and Intestinal Microflora of Chinese Mitten Crabs, Eriocheir sinensis.

Selenium is a vital trace mineral that is crucial for maintaining regular biological processes in aquatic animals. In this study, a four-week dietary trial was carried out to assess the impact of bio-fermented selenium (Bio-Se) on the growth and immune response of Chinese mitten crabs, Eriocheir sinensis. The crabs were randomly allocated to five dietary treatment groups, each receiving a different dose of Bio-Se. The doses included 0, 0.3, 0.6, 1.5, and 3.0 mg/kg and were accurately measured in basal diet formulations. The results showed the weight gain rate (WGR), specific growth rate (SGR), and survival rate (SR) in the 1.5 mg/kg Bio-Se group were the highest, and 3.0 mg/kg of Bio-Se has an inhibitory effect on the WGR, SGR, and SR. The activities of the immune enzymes, including glutathione peroxidase (GPX), superoxide dismutase (SOD), and acid phosphatase (ACP), of the hepatopancreas were significantly (p < 0.05) increased in the 1.5 mg/kg Bio-Se group, while they decreased (p < 0.05) in the 3.0 mg/kg feeding group compared to the 0 mg/kg feeding group. The concentration of maleic dialdehyde (MDA) exhibited the opposite pattern. Similarly, the mRNA expression levels of antimicrobial peptides (ALF-1, Crus-1, and LYS), ERK, and Relish genes were also observed to be the highest in the 1.5 mg/kg Bio-Se group compared with the other groups. Furthermore, the administration of 1.5 mg/kg of Bio-Se resulted in an increase in the thickness of the intestinal plica and mucosal layer, as well as in alterations in the intestinal microbial profile and bacterial diversity compared to the dose of 0 mg/kg of Bio-Se. Notably, the population of the beneficial bacterial phylum Fusobacteria was increased after crabs were fed the 1.5 mg/kg Bio-Se diet. In conclusion, the oral administration of 1.5 mg/kg of Bio-Se improved the growth efficiency, antioxidant capabilities, immunity, and intestinal health of E. sinensis. Through a broken-line analysis of the WGR against dietary Bio-Se levels, optimal dietary Bio-Se levels were determined to be 1.1 mg/kg. These findings contribute valuable insights to the understanding of crab cultivation and nutrition.

Preparation of PBAT microplastics and their potential toxicity to zebrafish embryos and juveniles

The extensive use of traditional non-biodegradable plastics results in the generation of microplastics (MPs), forming a new pollutant that can pose significant environmental risks. Biodegradable plastics (BP) possess degradation properties and can partially replace conventional plastics, thereby reducing pollution. However, further investigation is needed into the toxicity of biodegradable microplastics (BMPs) on aquatic organisms. This study explores the toxic effects of PBAT microplastics (PBAT-BMPs) and microplastics produced from degradable PBAT/TPS (thermoplastic starch) composite film (PBAT/TPS-BMPs) on zebrafish embryos. Our findings indicate that the presence of microplastics on the embryo's surface increases with higher BMP concentration. Nonetheless, PBAT-BMPs tend to aggregate and are blocked by the embryonic membrane, thus diminishing their toxic effects on the embryo. Acute toxicity experiments revealed that 30 mg/L of PBAT-BMPs significantly reduced the survival rate of zebrafish embryos, whereas PBAT/TPS-BMPs had a lesser effect on survival. Both types of BMPs influenced the hatching rate of the embryos, leading to prolonged incubation periods. Additionally, both types of BMPs impacted the locomotor behavior of zebrafish larvae, causing an increase in larval locomotor speed. However, these BMPs had little impact on larval body development and heartbeat behavior. Fluorescent microplastic tracer experiments demonstrated that PBAT-BMPs persisted in juvenile fish for at least 144 hours and were difficult to metabolize and excrete. Our study aims to gain a better understanding of the potential effects of BMPs on aquatic ecosystems and biological health, as well as to propose effective strategies for reducing environmental pollution and protecting organisms.

Genome-wide identification of crustacyanin and function analysis of one isoform high-expression in carapace from Neocaridina denticulata sinensis

Lipocalin proteins transport hydrophobic molecules, including apolipoprotein D, retinol-binding protein, and crustacyanin (CRCN). CRCN can combine with astaxanthin to cause a bathochromic shift in the emission spectrum of astaxanthin from red to blue. Therefore, CRCN influences the colors and patterns of crustaceans, which are important for various biological functions such as camouflage, reproduction, and communication. For aquatic organisms, body color is economically important and can be indicative of habitat water quality. In this study, thirteen CRCN genes (NdCRCNs) were first discovered in Neocaridina denticulata sinensis, contradicting prior findings of a few isoform genes in a species. The expression pattern of NdCRCNs in tissues showed that the expression of one CRCN isoform gene, named NdCRCN-30, was the highest in the carapace. In situ hybridization (ISH) analysis revealed that NdCRCN-30 was predominantly distributed in the outer epidermis of shrimp. Interference of NdCRCN-30 could cause a change in the color of the carapace. RNA-seq was performed after knockdown with the NdCRCN-30, and differential gene enrichment analysis revealed that this gene is primarily associated with antioxidant function, pigmentation, and molting. Overall, our results will provide new insights into the biological function of the CRCN and genetic breeding for changing body color in economic crustaceans.

Polystyrene nanoplastics significantly facilitate largemouth bass ranavirus infection of host cells

Novel pollutants nanoplastics (NPs) are widely distributed in aquatic environments and may pose a health threat to aquatic organisms. Notably, the contribution of NPs to the occurrence of viral diseases in aquatic animals remains largely uncertain. In this study, the effects of polystyrene nanoplastics (PS-NPs) on Largemouth bass ranavirus (LMBV)-infected MsF cells were investigated. MsF cells took up PS-NPs in a time- and dose-dependent manner and significantly affect cell viability at an exposure concentration of 500μg/mL. Western blot and qPCR assays indicated that exposure to PS-NPs accelerated LMBV replication in MsF cells. PS-NPs act synergistically with LMBV to disrupt the cellular antioxidant system, as evidenced by increased ROS production and decreased mRNA levels of antioxidant-associated genes. Furthermore, PS-NPs was found to exacerbate LMBV-induced inflammatory responses, as demonstrated by disturbed expression of inflammation-related factors. In addition, our results suggest that PS-NPs reduce IFN production by inhibiting the expression of molecules related to the cGAS-STING signaling pathway, thereby promoting viral replication. Collectively, our findings suggest the potential threat of NPs to infectious diseases caused by freshwater fish viruses and provide new insights for fish disease prevention and control.