Search

The large yellow croaker (Larimichthys crocea) displays pronounced behavioral changes in response to low oxygen environments, including restlessness, constant floating, and increases in respiratory rate and tail beat frequency. However, the underlying molecular mechanisms governing these behavioral and physiological alterations remain unclear. To address this knowledge gap, the present study employed transcriptome analysis to investigate the molecular responses in the large yellow croaker muscle tissue after the fish were exposed to hypoxic treatment. A total of 4553, 1797, 3222, 3148, and 4815 differentially expressed genes (DEGs) were identified at 1 h, 6 h, 9 h, 24 h, and 72 h after hypoxia treatment, respectively. Subsequent analysis utilizing Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichments unveiled a notable up-regulation of genes related to muscular movement-related pathways, specifically the calcium pathway. This up-regulation coincided with an observed increase in the intracellular Ca2+ concentration and Ca2+ transport enzyme activity, suggesting that hypoxia-induced muscle contraction and relaxation occurred by elevating the intracellular Ca2+ concentration within the muscle tissue. Key DEGs associated with the glycolysis and the TCA cycle pathways were also found to be significantly increased in expression, indicating an activation of energy generation pathways in the muscle of hypoxic large yellow croakers. Conversely, genes related to protein synthesis and cell growth were significantly down-regulated, suggesting an inhibition of energy-consuming processes. Additionally, hypoxia was observed to impact various physiological processes such as oxygen transport, vascular control, and apoptosis. These processes may have been mediated by the up-regulation of HIF-1α and its target genes. Consequently, these results offer novel insights into comprehending the molecular basis of the changes in behavior and physiology of large yellow croaker in response to the stress induced by low oxygen levels.

Pseudomonas plecoglossicida is a major pathogen that causes visceral white nodules disease (VWND), a highly devastating disease, in cultured large yellow croaker (Larimichthys crocea), leading to severe economic losses. Peptide chain release factor methyltransferase hemK in bacteria plays a role in pathogenicity, but its exact function in P. plecoglossicida remains unclear. In this study, a P. plecoglossicida mutant strain (ΔhemK) was constructed by deleting hemK, and the roles of hemK in regulating P. plecoglossicida virulence were thereafter investigated. Compared to the wildtype strain, cell motility and biofilm formation of ΔhemK were weaken, and extracellular protein profile was altered. In vitro study showed that the hemK deletion significantly decreased the survival rate of P. plecoglossicida in large yellow croaker macrophage cell line (LYC-FM) by impairing oxidative stress tolerance. Furthermore, large yellow croakers infected with ΔhemK showed a significant reduction in mortality rate, bacterial load, and noticeable nodules within the spleen, when compared to the wild-type strain. These findings indicated that hemK plays an essential role in the pathogenicity of P. plecoglossicida, thus providing a valuable insight into the pathogenic mechanisms of P. plecoglossicida and offering a potential target for its attenuated-vaccine development.

Clostridioides difficile, the etiological agent of C. difficile infection (CDI), elicits a spectrum of diarrheal symptoms with varying severity and the potential to result in severe complications such as colonic perforation, pseudomembranous colitis, and toxic megacolon. The perturbation of gut microbiome, often triggered by antibiotic usage, represents the primary factor augmenting the risk of CDI. This underscores the significance of interactions between C. difficile and the microbiome in determining pathogen adaptability. In recent years, researchers have increasingly recognized the pivotal role played by intestinal microbiota in host health and its therapeutic potential as a target for medical interventions. While extensive evidence has been established regarding the involvement of gut bacteria in CDI, our understanding of symbiotic interactions between hosts and fungi within intestinal microbiota remains limited. Herein, we aim to comprehensively elucidate both composition and key characteristics of gut fungal communities that significantly contribute to CDI, thereby enhancing our comprehension from pharmacological and biomarker perspectives while exploring their prospective therapeutic applications for CDI.

Rab7 is a small GTPase that regulates vesicular traffic from early to late endosomal stages of the endocytic pathway. In this study, full-length cDNA sequences of Rab7 were cloned from Yellow River Carp Cyprinus carpio, which were designated as CcRab7, and its function during pathogen infection was also investigated. The results showed that the full-length cDNA of CcRab7 was composed of 1511 bp, and open reading frame is 618 bp encoding 205 amino acids with 23.10 kDa. The deduced amino acid sequence of CcRab7 shared high similarity to that of Pimephales promelas (99.02%). In this paper, real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression levels in 11 tissues and organs of healthy Yellow River Carp, and the tissue distribution of CcRab7 as well as its defense mechanism against Spring viremia of carp virus (SVCV) and Aeromonas hydrophila in Yellow River Carp C. carpio was investigated. The results revealed that the mRNA transcripts of CcRab7 were widely detected in various tissues of C. carpio, with the highest expression levels in head kidney and blood, indicating that CcRab7 was involved in the innate immune process. Yellow River Carp was immunized with A. hydrophila and SVCV respectively, and the expression changes of CcRab7 were detected in gill, spleen, liver and skin of Yellow River Carp. The mRNA transcripts of CcRab7 in the main organs (gill, and liver, skin and spleen) were significantly affected by SVCV and A. hydrophila infection. The results showed that the CcRab7 expression levels was obviously up-regulated at different time points. The eukaryotic recombinant plasmids of CcRab7 and pEGFP-N3 transfected into Grass carp ovary cell lines (GCO) cells for subcellular localization. The results provide a basis for further investigation into the immunological role of CcRab7 in Yellow River Carp.

The Gram-negative bacterium Pseudomonas plecoglossicida has caused visceral granulomas disease in several farmed fish species, including large yellow croaker (Larimichthys crocea), which results in severe economic losses. Type III secretion systems (T3SS) are protein secretion and translocation nanomachines widely employed by many Gram-negative bacterial pathogens for infection and pathogenicity. However, the exact role of T3SS in the pathogenesis of P. plecoglossicida infection is still unclear. In this study, a T3SS translocators deletion strain (△popBD) of P. plecoglossicida was constructed to investigate the function of T3SS. Then comparative secretome analysis of the P. plecoglossicida wild-type (WT) and △popBD mutant strains was conducted by label-free quantitation (LFQ) mass spectrometry. The results show that knockout of T3SS translocators popB and popD has an adverse effect on the effector protein ExoU secretion, flagella assembly, and biofilm formation. Further experimental validations also confirmed that popB-popD deletion could affect the P. plecoglossicida flagella morphology/formation, adherence, mobility, and biofilm formation. These data indicate that a cross-talk exists between the P. plecoglossicida T3SS and the flagella system. Our results, therefore, will facilitate the further under-standing of the pathogenic mechanisms leading to visceral granulomas disease caused by P. plecoglossicida.

Acidifier has become one of the most important substitutes for antibiotics, and its beneficial effects on growth performance and digestive ability have been evidenced. Sodium benzoate is a commonly used acidifier. A 56-day feeding test was carried out to elucidate the effects of dietary sodium benzoate on growth performance, morphology, antioxidant activity and intestinal microflora of juvenile Litopenaeus vannamei with an average initial body weight of 0.55 ± 0.01 g. Six experimental diets were prepared by adding 0, 750, 1000, 1250, 1500 and 1750 mg/kg sodium benzoate to the control diet. The results showed that: The final body weight (FBW) and weight gain (WG) of juvenile shrimp were significantly improved by diet supplemented with 1750 mg/kg sodium benzoate. Dietary 1750 mg/kg sodium benzoate significantly increased the trypsin activity. The activities of catalase (CAT), superoxide dismutase (SOD) and glutathione S-transferase (GST) in the intestine and hepatopancreas of the 1750 mg/kg sodium benzoate group were markedly higher than those of the control group. Furthermore, the expression levels of trypsin (TRY), amylase (AMS), lysozyme (LZM), interleukin-8 (IL8) and interleukin-10 (IL10) genes in the intestine significantly increased when the content of sodium benzoate in the diet was greater than or equal to 1000 mg/kg. The addition of 1500 and 1750 mg/kg sodium benzoate to the feed improved the intestinal and hepatopancreatic tissue structures of juvenile shrimp and enriched the diversity of intestinal microbiota. Sodium benzoate residues were not detected in each sodium benzoate supplemented group. At the end of the Aeromonas hydrophila challenge test, juvenile shrimp in the 1500 and 1750 mg/kg group exhibited significantly higher cumulative survival rates than that in the control group. Conclusively, dietary 1750 mg/kg sodium benzoate could improve the growth performance, antioxidant capacity, morphology, intestinal microflora and disease resistance of juvenile shrimp.

Background and aims: Clostridioides difficile infection (CDI) induces intense acute inflammatory responses through toxin release. A combination of antibiotic and anti-inflammatory agents is sometimes recommended in severe, non-responsive cases, although clinical trials have been inconclusive, raising concerns about potential complications. This study aims to investigate the effect of budesonide and mesalamine in the treatment of CDI in a murine model, by evaluating the combination of fidaxomicin and these anti-inflammatory drugs. Method: C57BL/6 J female mice pretreated with an antimicrobial mixture were challenged with C. difficile VPI 10463 or culture media by gavage. After the challenge, mice received placebo, fidaxomicin alone (20 mg/kg), or fidaxomicin combined with mesalamine (200, 400 mg/kg) or budesonide (0.2, 1, 10 mg/kg) for 5 days. The mice were monitored for 7 days with weight and survival. Colon and cecum tissues were harvested for histological assessment. Results: CDI of mice caused 80% mortality. Fidaxomicin completely protected against CDI in all parameters (weight, survival and pathscores). Mortality rates were up to 90%, 70% in budesonide(10 mg/kg) and mesalamine (400 mg/kg) treatment group, respectively. Budesonide (0.02,0.1 and 1 mg/kg) adjunction to fidaxomicin worsened the disease outcome according to all tested parameters. While mesalamine in combination with fidaxomicin (200, 400 mg/kg) did not lead to any deaths during CDI treatment, it did not provide additional benefits. Conclusions: Anti-inflammatory drugs including corticosteroid therapy may worsen the incidence and severity of CDI in this mouse model. These studies may have important clinical implications for understanding the role of anti-inflammatory/ corticosteroid therapy in CDI and inflammatory bowel disease management.

Dimethyl disulfide (DMDS) is a relatively new soil fumigant used in agro-industrial crop production to control soil-borne pests that damage crops and reduce yield. The emissions of DMDS after fumigation reduce soil concentrations thus reducing the risk of phytotoxicity to newly planted crops. However, the factors affecting the desorption of DMDS from soil are unclear. In our study, the desorption characteristics of DMDS from soil were measured in response to continuous ventilation. The degradation of DMDS in soil was examined by thermal incubation. The phytotoxic response of newly-planted cucumber (Cucumis sativus) seedlings to DMDS residues was measured by a sand culture experiment. The results showed DMDS desorption and degradation rates fit a first-order model; that 92% of the DMDS desorption occurred in the first hour after fumigant application; and that residue concentrations in the soil at the end of the ventilation period were unlikely to be phytotoxic to newly-planted cucumber seedlings. By the third day of ventilation, the average desorption rate (ADR) of DMDS in Wenshan soil was 4.0 and 3.6 times, respectively, faster than that in Shunyi and Suihua soils and the ADR of DMDS in soil decreased by 40.0% when the soil moisture content increased from 3% to 12% (wt/wt). Moreover, within one hour of ventilation, the ADR of DMDS in soil decreased by 20.1% when the soil bulk density increased from 1.1 to 1.3 g cm-3. The degradation of DMDS in soil, however, was mostly influenced by soil type and moisture content. A slow degradation rate resulted in a high initial desorption concentration of DMDS in soil. Our results indicated that DMDS desorption from soil in response to continuous ventilation was affected by the soil type, moisture content and bulk density. Rapid degradation of DMDS in soil will lower the risk of phytotoxic residues remaining in the soil and reduce emissions during the waiting period. Acceleration of emissions early in the waiting period by managing soil moisture content or increasing soil porosity may shorten the duration of emissions. Alternatively, soil extraction technology could be developed to recover and reduce fumigant emissions.

An 8 weeks feeding trail was conducted to evaluate the potential of poultry by-product meal (PBM) replacing fish meal (FM) in the diet of juvenile large yellow croaker (Larimichthys crocea). Five iso-nitrogenous (45% crude protein), iso-lipidic (10% crude lipid) and iso-energetic (about kcal 400 g diet −1) diets were formulated with 0, 25%, 50% 75% and 100% FM being replaced by PBM (marked as FM, PBM25, PBM50, PBM75 and PBM100). Crystal methionine and lysine were added into PBM-containing diets to match its concentration in FM diet. Each diet was randomly assigned to triplicate groups of 30 fish (initial weight 18.43 ± 1.28 g) in floating sea cages. The weight gain, specific growth rate and feed efficiency of L. crocea decreased significantly when the replacement level of FM was higher than 25%. The feed intake of fish fed PBM100 was significantly lower than those fed FM, PBM25 and PBM50. Juveniles fed PBM-containing diets showed significantly lower value of muscular thickness in intestine when compared to those fed FM. Villi length decreased significantly when the replacement of FM in diets was more than 50%. Fish fed FM diet had the relative higher abundances of Exiguobacterium (from order to genus), Bacillus (from order to genus), Pseudomonadales (from order to genus) and Acinetobacter genus, those fed PBM25, PBM50 and/or PBM75 showed higher abundance of Peptostreptococcaceae and Vibrio. Overall, PBM replacing dietary FM may inhibit growth performance of L. crocea through reducing digestibility via declining the abundance of intestinal bacteria that producing digestive enzyme, and improving the abundance of pathogen. Peptostreptococcaceae and Vibrio could be the indicator in the intestinal of L. crocea fed high PBM diets. The optimum dietary FM replacement level by PBM was 25% for L. crocea.

Summary: How organisms cope with coldness and high pressure in the hadal zone remains poorly understood. Here, we sequenced and assembled the genome of hadal sea cucumber Paelopatides sp. Yap with high quality and explored its potential mechanisms for deep-sea adaptation. First, the expansion of ACOX1 for rate-limiting enzyme in the DHA synthesis pathway, increased DHA content in the phospholipid bilayer, and positive selection of EPT1 may maintain cell membrane fluidity. Second, three genes for translation initiation factors and two for ribosomal proteins underwent expansion, and three ribosomal protein genes were positively selected, which may ameliorate the protein synthesis inhibition or ribosome dissociation in the hadal zone. Third, expansion and positive selection of genes associated with stalled replication fork recovery and DNA repair suggest improvements in DNA protection. This is the first genome sequence of a hadal invertebrate. Our results provide insights into the genetic adaptations used by invertebrate in deep oceans.

Tris (2-chloroethyl) phosphate (TCEP), an emerging environmental pollutant, has been frequently detected in natural waters. The objective of this study was to investigate possible parental transfer of TCEP and transgenerational effects on the early development and thyroid hormone homeostasis in F1 larvae following parental whole life-cycle exposure to TCEP. To this end, zebrafish (Danio rerio) embryos were exposed to environmentally relevant concentrations (0.8, 4, 20 and 100 μg/L) of TCEP for 120 days until sexual maturation. Parental exposure to TCEP resulted in significant levels of TCEP, developmental toxicity including decreased survival and final hatching rates, accelerated heart rate and elevated malformation rate, as well as induction of oxidative stress and cell apoptosis in F1 offspring. In F1 eggs, declined thyroxin (T4) levels were observed, consistent with those in plasma of F0 adult females, indicating the maternal transfer of thyroid endocrine disruption to the offspring. In addition, mRNA levels of several genes along the hypothalamic-pituitary-thyroid (HPT) axis were significantly modified in F1 larvae, which could be linked to transgenerational developmental toxicity and thyroid hormone disruption. For the first time, we revealed that the parental exposure to environmentally relevant levels of TCEP could cause developmental toxicity and thyroid endocrine disruption in subsequent unexposed generation.

Hypoxia occurrence is potentiated by eutrophication caused by human activities and it may be the most dramatically increasing environmental factor as a result of the climate change-led increasing temperatures. The crucian carp Carassius auratus is an important aquaculture economical freshwater species and is also a potential model for the study of physiological adaptation. However, the transcriptomics and genomics data available for this species are still scarce. The gills of fish are the major organ of hypoxia and other stresses. In this study, we studied the effect of the exposure to hypoxia (1.0 mg O2/L for 7 d) and 2 mg/mL Poly I:C on the gene expression of the gills of Carassius auratus using the Illumina sequencing platform. Comparison of the gene expression levels between the experimental and control groups showed differentially expressed genes (DEGs). The DEGs were mainly annotated into glycolysis, oxidative phosphorylation and immune system categories based on the KEGG database. In glycolysis pathway, lactate dehydrogenase, malate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase, enolase and succinate dehydrogenase were identified. In addition, the DEGs were enriched in the immune-related pathway functions, including pattern recognition receptor (PRR), antigen processing and presentation, B cell and T cell receptor signaling pathways, natural killer (NK) cell mediated cytotoxicity, Fc gamma R-mediated phagocytosis, and the JAK-STAT, p53 and cytokines signaling pathways. This study also produced potential hypoxia biomarkers including FOS, JUN B, lysosomal acid lipase, and HIF-1α. HIF-1 signaling pathway was significantly down-regulated after hypoxia and Poly I: C exposure. Hence, the data generated here may offer a valuable resource information for the further study of the immune response and also provide improved strategies against hypoxia and Poly I:C in teleost fish.

Interleukin-2 (IL-2) signals influence various lymphocyte subsets during differentiation, immune responses and homeostasis. IL-2 acts on different cells by binding to its receptors (IL-2R), which consists of three subunits, IL-2Rα (CD25), IL-2Rβ (CD122), and the common gamma chain or γc (CD132). In the present study, three IL-2 receptor subunits, designated as LcCD25-like (LcCD25L), LcIL-2Rβ and Lcγc, were characterized in large yellow croaker (Larimichthys crocea). The LcCD25L, like other teleost CD25L or IL-2/IL-15Rα, contains only one sushi domain at N-terminus. The synteny of CD25L from different teleost are conserved. The deduced protein of LcIL-2Rβ and Lcγc exhibits a typical class I cytokine receptors architecture, including a cytokine-binding homology domain (CHD) consisting of two fibronectin type-III (FNIII) domains (D1 and D2) and a conserved WSXWS motif in D2 domain. These three IL-2 receptor subunits were constitutively expressed in all tissues and primary immune-related cells examined. The LcCD25L was highly expressed in blood, while LcIL-2Rβ and Lcγc were highly expressed in spleen and gill. For immune-related cells, LcCD25L and LcIL-2Rβ were highly expressed in PKLs, while the Lcγc exhibited the highest expression in PKMs. These three IL-2 receptor subunits could be dramatically induced by T cell mitogen PHA in PKLs, which mainly composed of T and B lymphocytes. The results presented indicated that large yellow croaker IL-2R might exercise function on lymphocytes, especially on activated T cells.

Background: Hydatid disease is a serious parasitic disease threatening public health. Because of its rarity in non-endemic coastal areas, determining the nature and origin of a chronic, enlarged liver cystic mass is challenging in these regions. Under these circumstances, physicians need a confirmatory diagnostic tool beyond immunological and radiological examinations. This study investigated a novel human single-chain fragment variable (scFv) antibody for the confirmative diagnosis of 18 atypical hydatid disease cases in non-endemic coastal areas. Results: A scFv antibody against cystic echinococcosis was produced by genetic engineering and then applied to the immunohistochemical diagnosis of 18 cases of cystic echinococcosis presented in non-endemic coastal areas. The diagnosis of these cases by ultrasound and serum-based examinations was inconclusive. The 750 bp scFv antibody gene was expressed in COS-7 cells, and the antibody localized in the cytoplasm. The scFv antibody can detect the germinal layer and protoscolices of actively growing cysts but not of the degenerating protoscolices and has a diagnostic efficiency higher than that of single serum or ultrasound testing (P

The dinoflagellate Noctiluca scintillans is often reported as a worldwide HAB species and caused severe financial losses to local aquaculture. In this review, we summarized the temporal and geographical distribution of its HABs in China, as well as its position in the plankton structure. Increasing N. scintillans HABs, both frequency and coverage, have broken out in almost all Chinese coastal regions mainly from April to June, with short-term and small coverage blooms as the primary type. The HAB period seems to shift with age and latitude. Recently, increasing abundance and dominance of N. scintillans were also reported in plankton communities in Chinese coastal waters, with multiple environmental factors related. In particular, trophic relationships may play an important role in its dominance and outbreaks of HABs. However, how N. scintillans became a dominant species in China and the mechanisms responsible for its HABs require further study., 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.)