Studies on immunological characteristics and transcriptomic analysis of Litopenaeus vannamei low salt-tolerance family.

Litopenaeus vannamei is a widely distributed euryhaline aquatic animal, affected by low salinity, which can impact its disease resistance and immunity. However, there is a limited understanding of the adaptation mechanisms of L. vannamei with different genetic backgrounds to low salinity. Therefore, the present study aimed to compare the immunity characteristics and transcriptomics of L. vannamei low salt-tolerant (FG I/J) and low salt-sensitive (control) families. Also, the disease resistance and immune parameters (including [THC], hemolymph cell viability, lysozyme activity [LZM], phenoloxidase content [PO], interleukin-6 [IL-6], and tumor necrosis factor-alpha [TNF-α]) of the FG I/J and control families of L. vannamei under low salinity (5‰) and ambient salinity (24‰) were examined. Additionally, hepatopancreas transcriptomics of the FG I/J and control families were analyzed at a salinity of 5‰. The results showed that the FG I/J family had higher disease resistance to Vibrio parahaemolyticus and stronger immunological capacity than the control family. Transcriptomic analysis showed significantly enriched energy metabolism and immune regulation pathways. Therefore, we speculated that energy metabolism provides sufficient energy for immunological modulation in the FG I/J family to deal with long-term low-salt stress and achieve high growth and survival rates.

Olfactory toxicity of tetrabromobisphenol A to the goldfish Carassius auratus.

Tetrabromobisphenol A (TBBPA) is one of the most extensively used brominated flame retardants and its increasing use in consumer products has raised concerns about its ecotoxicity. Given the ubiquity of TBBPA in aquatic environments, it is inevitable that these chemicals will enter the olfactory chambers of fish via water currents. Nevertheless, the olfactory toxicity of TBBPA to aquatic organisms and the underlying toxic mechanisms have yet to be elucidated. Therefore, we investigated the olfactory toxicity of TBBPA in the goldfish Carassius auratus, a model organism widely used in sensory biology. Results showed that exposure to TBBPA resulted in abnormal olfactory-mediated behaviors and diminished electro-olfactogram (EOG) responses, indicating reduced olfactory acuity. To uncover the underlying mechanisms of action, we examined the structural integrity of the olfactory epithelium (OE), expression levels of olfactory G protein-coupled receptors (GPCRs), enzymatic activities of ion transporters, and fluctuations in neurotransmitters. Additionally, comparative transcriptomic analysis was employed to investigate the molecular mechanisms further. Our study demonstrates for the first time that TBBPA at environmentally relevant levels can adversely affect the olfactory sensitivity of aquatic organisms by interfering with the transmission of aqueous stimuli to olfactory receptors, impeding the binding of odorants to their receptors, disrupting the olfactory signal transduction pathway, and ultimately affecting the generation of action potentials.

Micro/nanoplastics impair the feeding of goldfish by disrupting the complicated peripheral and central regulation of appetite.

The ubiquitous presence of plastic particles in water bodies poses a potential threat to aquatic species. Although numerous adverse effects of microplastics (MPs) and nanoplastics (NPs) have been documented, their effects on fish feeding, one of the most important behaviors of animals, are far from being fully understood. In this study, the effects of MPs and NPs (at environmentally realistic levels) on fish food consumption and feeding behavior were assessed using goldfish (Carassius auratus) and polystyrene (PS) particles as representatives. In addition, to reveal the potential mechanisms, the effects of MPs and NPs on peripheral and central regulation of appetite were evaluated by examining appetite-regulation related intestinal, serous, and hypothalamic parameters. The results obtained indicated that the 28-day MP- and NP-exposure significantly impaired goldfish feeding by disrupting peripheral and central appetite regulation. Based on differences observed in their effects on the abovementioned behavioral, histological, and physiological parameters, MPs and NPs may interfere with appetite regulation in a size-dependent manner. Blocking the gastrointestinal tract and causing histopathological and functional damage to inner organs may be the main routes through which MPs and NPs disrupt appetite regulation. Our findings suggested that plastic particles exposure may have far-reaching effects on fish species through impaired feeding, which warrants further attention.

Microplastics weaken the exoskeletal mechanical properties of Pacific whiteleg shrimp Litopenaeus vannamei.

The ubiquitous presence of microplastics (MPs) in aquatic environments poses a significant threat to crustaceans. Although exoskeleton quality is critical for crustacean survival, the impact of MPs on crustacean exoskeletons remains elusive. Our study represents a pioneering effort to characterize the effects of MPs exposure on crustacean exoskeletons. In this study, the mechanical properties of whiteleg shrimp Litopenaeus vannamei exoskeletons were analyzed after exposure to environmentally realistic levels of MPs. Nanoindentation data demonstrated that MPs exposure significantly increased the hardness and modulus of both the carapace and abdominal segments of L. vannamei. Moreover, fractures and embedded MPs were detected on the exoskeleton surface using SEM-EDS analysis. Further analysis demonstrated that the degree of chitin acetylation (DA) in the shrimp exoskeleton, as indicated by FTIR peaks, was reduced by MPs exposure. In addition, exposure to MPs significantly inhibited the muscle Ca2+-ATPase activity and hemolymph calcium levels. Transcriptome and metabolome analyses revealed that the expression levels of genes encoding key enzymes and metabolites in the chitin biosynthetic pathway were significantly affected by MPs exposure. In conclusion, MPs at environmentally relevant concentrations may affect the exoskeletal mechanical properties of L. vannamei through a comprehensive mechanism involving the disruption of the crystalline structure of chitin, assimilation into the exoskeleton, and dysregulation of exoskeleton biosynthesis-related pathways.

Exposure to Polystyrene Nanoplastics Led to Learning and Memory Deficits in Zebrafish by Inducing Oxidative Damage and Aggravating Brain Aging.

Nanoplastics (NPs) may pass through the blood-brain barrier, giving rise to serious concerns about their potential toxicity to the brain. In this study, the effects of NPs exposure on learning and memory, the primary cognitive functions of the brain, are assessed in zebrafish with classic T-maze exploration tasks. Additionally, to reveal potential affecting mechanisms, the impacts of NPs exposure on brain aging, oxidative damage, energy provision, and the cell cycle are evaluated. The results demonstrate that NP-exposed zebrafish takes significantly longer for their first entry and spends markedly less time in the reward zone in the T-maze task, indicating the occurrence of learning and memory deficits. Moreover, higher levels of aging markers (ß-galactosidase and lipofuscin) are detected in the brains of NP-exposed fish. Along with the accumulation of reactive free radicals, NP-exposed zebrafish suffer significant levels of brain oxidative damage. Furthermore, lower levels of Adenosine triphosphate (ATP) and cyclin-dependent kinase 2 and higher levels of p53 are observed in the brains of NP-exposed zebrafish, suggesting that NPs exposure also results in a shortage of energy supply and an arrestment of the cell cycle. These findings suggest that NPs exposure may pose a severe threat to brain health, which deserves closer attention.

Identification, characterization, and agglutinating activity of a novel C-type lectin domain family 3 member B (CLEC3B) discovered in golden pompano, Trachinotus ovatus.

The lectins are a large family of carbohydrate-binding proteins that play important roles in the innate immune response of various organisms. Although C-type lectin domain family 3 member B (CLEC3B), an important member of C-type lectin, has been well documented in humans and several other higher vertebrates, little is currently known about this molecule in economically important marine fish species. In this study, through transcriptomic and BLAST screening, a novel CLEC3B gene was identified in the golden pompano (Trachinotus ovatus). The T. ovatus CLEC3B (ToCLEC3B) was subsequently characterized by bioinformatic analysis and compared with those reported in other species. In addition, the expression patterns of ToCLEC3B in different tissues under normal condition and at different times post pathogen challenge were assessed. Furthermore, the agglutinating activity of ToCLEC3B with and without Ca2+ against different bacteria and blood cells of donor species were verified using the recombinant T. ovatus CLEC3B (rToCLEC3B). Our results demonstrated that ToCLEC3B is a Ca2+-dependent galactose-binding lectin with a single copy of carbohydrate recognition domain (CRD). Similar to CLEC3B reported in other species, the CRD domain of ToCLEC3B consists of two α-helices, six ß-sheets, and four loops, forming two Ca2+- and a galactose-binding sites. According to the phylogenetic analysis, the ToCLEC3B was highly similar (similarity at 95.00%) to that of its relative, the greater amberjack (Seriola dumerili). The expression of ToCLEC3B was detected in all tissues examined under normal condition and was significantly up-regulated by injection of pathogenic microbes. In addition, the rToCLEC3B exhibited strong agglutinating activity against different bacteria and blood cells of donor species in the presence of Ca2+. Our results indicate that ToCLEC3B is a constitutive and inducible acute-phase immune factor in the host's innate immune response of T. ovatus.

Toxic effects of four emerging pollutants on cardiac performance and associated physiological parameters of the thick-shell mussel (Mytilus coruscus).

Robust cardiac performance is critical for the health and even survival of an animal; however, it is sensitive to environmental stressors. At present, little is known about the cardiotoxicity of emerging pollutants to bivalve mollusks. Thus, in this study, the cardiotoxic effects of four emergent pollutants, carbamazepine (CBZ), bisphenol A (BPA), tetrabromobisphenol A (TBBPA), and tris(2-chloroethyl) phosphate (TCEP), on the thick-shell mussel, Mytilus coruscus, were evaluated by heartbeat monitoring and histological examinations. In addition, the impacts of these pollutants on parameters that closely related to cardiac function including neurotransmitters, calcium homeostasis, energy supply, and oxidative status were assessed. Our results demonstrated that 28-day exposure of the thick-shell mussel to these pollutants resulted in evident heart tissue lesions (indicated by hemocyte infiltration and myocardial fibrosis) and disruptions of cardiac performance (characterized by bradyrhythmia and arrhythmia). In addition to obstructing neurotransmitters and calcium homeostasis, exposure to pollutants also led to constrained energy supply and induced oxidative stress in mussel hearts. These findings indicate that although do differ somehow in their effects, these four pollutants may exert cardiotoxic impacts on mussels, which could pose severe threats to this important species and therefore deserves more attention.

Toxicity of microplastics and triclosan, alone and in combination, to the fertilisation success of a broadcast spawning bivalve Tegillarca granosa.

Since most marine invertebrates adopted external fertilisation, their fertilisation process is particularly vulnerable to aquatic pollutants. Both antimicrobial ingredients and microplastics (MPs) are ubiquitous in aquatic environments; however, their synergistic effects on the fertilisation of marine invertebrates remain unclear. Therefore, in this study, the fertilisation toxicity of MPs and triclosan (TCS), alone and in combination, was investigated in the broadcast spawning bivalve Tegillarca granosa. Results showed that MPs and TCS significantly suppressed the fertilisation success of T. granosa. As the fertilisation success of broadcast spawning invertebrates depends on successful gamete collisions, gamete fusion, and egg activation, sperm swimming velocity, viability, gamete collision probability, ATP status, and ion-transport enzyme activities were also analysed to further ascertain the underlying toxicity mechanisms. In summary, our findings indicate that the presence of MPs may enhance the fertilisation toxicity of TCS by hampering sperm-egg collision probability, reducing gamete fusion efficiency, and restricting Ca2+ oscillation formation.

Tris(2-chloroethyl) Phosphate Exerts Hepatotoxic Impacts on Zebrafish by Disrupting Hypothalamic-Pituitary-Thyroid and Gut-Liver Axes.

The ubiquitous environmental presence of tris(2-chloroethyl) phosphate (TCEP) poses a potential threat to animals; however, little is known about its hepatotoxicity. In this study, the effects of TCEP exposure (0.5 and 5.0 µg/L for 28 days) on liver health and the potential underlying toxification mechanisms were investigated in zebrafish. Our results demonstrated that TCEP exposure led to hepatic tissue lesions and resulted in significant alterations in liver-injury-specific markers. Moreover, TCEP-exposed fish had significantly lower levels of thyrotropin-releasing hormone and thyroid-stimulating hormone in the brain, evidently less triiodothyronine whereas more thyroxine in plasma, and markedly altered expressions of genes from the hypothalamic-pituitary-thyroid (HPT) axis in the brain or liver. In addition, a significantly higher proportion of Bacteroidetes in the gut microbiota, an elevated bacterial source endotoxin lipopolysaccharide (LPS) in the plasma, upregulated expression of LPS-binding protein and Toll-like receptor 4 in the liver, and higher levels of proinflammatory cytokines in the liver were detected in TCEP-exposed zebrafish. Furthermore, TCEP-exposed fish also suffered severe oxidative damage, possibly due to disruption of the antioxidant system. These findings suggest that TCEP may exert hepatotoxic effects on zebrafish by disrupting the HPT and gut-liver axes and thereafter inducing hepatic inflammation and oxidative stress.

Impacts of ammonia stress on different Pacific whiteleg shrimp Litopenaeus vannamei families and the underlying adaptive mechanisms.

Ammonia stress in aquaculture systems poses a great threat to the growth and survival of the Pacific whiteleg shrimp Litopenaeus vannamei. Although the ammonia stress tolerance capacity of L. vannamei has been found to vary significantly among different breeding families, the underneath mechanisms are still largely unknown. In this study, the ammonia tolerance capacity of different L. vannamei breeding families was compared. Results confirmed the significant differences in the ammonia adaptability among different families. To ascertain the underlying adaptive strategies, ATP status, ATP synthase activity, expression and activities of ammonia excretion and metabolism-related enzymes, and apoptosis in shrimp gills were analyzed. Furthermore, transcriptomic analyses were also performed to elucidate the molecular mechanisms. Our results indicated that ammonia-tolerant L. vannamei may possess (1) enhanced ability to excrete ammonia, (2) better capacity to convert ammonia into less toxic products, and (3) sufficient energy reserves for ammonia-compensating processes.

Triclosan and triclocarban weaken the olfactory capacity of goldfish by constraining odorant recognition, disrupting olfactory signal transduction, and disturbing olfactory information processing.

Recently, increased production and consumption of disinfectants such as triclosan (TCS) and triclocarban (TCC) have led to massive pollution of the environment, which draws global concern over the potential risk to aquatic organisms. However, the olfactory toxicity of disinfectants in fish remains elusive to date. In the present study, the impact of TCS and TCC on the olfactory capacity of goldfish was assessed by neurophysiological and behavioral approaches. As shown by the reduced distribution shifts toward amino acid stimuli and hampered electro-olfactogram responses, our results demonstrated that TCS/TCC treatment would cause deterioration of the olfactory ability of goldfish. Our further analysis found that TCS/TCC exposure suppressed the expression of olfactory G protein-coupled receptors in the olfactory epithelium, restricted the transformation of odorant stimulation into electrical responses by disturbing the cAMP signaling pathway and ion transportation, and induced apoptosis and inflammation in the olfactory bulb. In conclusion, our results demonstrated that an environmentally realistic level of TCS/TCC would weaken the olfactory capacity of goldfish by constraining odorant recognition efficiency, disrupting olfactory signal generation and transduction, and disturbing olfactory information processing.