Cellular responses in crustaceans under white spot syndrome virus infection.

Innate immunity plays the most important system responsible for protecting crustaceans against invading pathogens. White spot syndrome virus (WSSV) is considered a serious pathogen in crustaceans with high cumulative mortality and morbidity in infected animals. Understanding the mechanism of the response of hosts to WSSV infection is necessary, which is useful for effective prevention in controlling infection. In this review, we summarize the participation of signaling pathways (toll, immune deficiency, JAK/STAT, endocytosis, mitogen-activated protein kinase, PI3K/Akt/mTOR, cGAS-STING, Wingless/Integrated signal transduction, and prophenoloxidase (proPO) cascade) and the activity of cells (apoptosis, autophagy, as well as, reactive oxygen species and antioxidant enzymes) in the cellular-mediated immune response of crustaceans during WSSV infection. The information presented in this current review is important for a better understanding of the mechanism of the response of hosts to pathogens. Additionally, this provides a piece of basic knowledge for discovering approaches to strengthen the immune system and resistance of cultured animals against viral infections.

Identification and Characterization of a Potential Probiotic, Clostridium butyricum G13, Isolated from the Intestine of the Mud Crab (Scylla paramamosain).

The butyrate-producing bacterium Clostridium butyricum has been proven to be important in improving the growth and health benefits of aquatic animals. In this study, C. butyricum G13 was isolated for the first time from the gut of the mud crab (Scylla paramamosain). The results of this study showed that C. butyricum G13 could produce a high concentration of butyric acid and grow well in a wide range of pHs (4 to 9) and NaCl (1 to 2.5%) and bile salt (0.2 to 1.0%) concentrations. In vitro characterization revealed that C. butyricum G13 is a Gram-positive and gamma-hemolytic bacterium sensitive to most antibiotics and shows hydrophobicity and the capacity to degrade starch. In vitro fermentation using mud crab gut contents showed that C. butyricum G13 alone or in combination with galactooligosaccharides (GOS) and/or resistant starch (RS) significantly increased butyric acid production and beneficially affected the abundance and diversity of intestinal microbiota. In addition, C. butyricum G13 can improve the survival rate of mud crabs and effectively maintain the normal structure of gut morphology after infection with Vibrio parahaemolyticus. In conclusion, C. butyricum G13 can be considered a potential probiotic that improves the immune capacity and confers health benefits on mud crabs. IMPORTANCE With the development of society, more and more aquatic animals are demanded. Intensification in the aquaculture scale is facing problems, such as disease outbreaks, eutrophication of water bodies, and misuse of antibiotics. Among these challenges, disease outbreak is the most important factor directly affecting aquaculture production. It is crucial to explore new approaches effective for the prevention and control of diseases. Probiotics have been widely used in aquaculture and have shown beneficial effects on the host. In this study, the butyrate-producing bacterium Clostridium butyricum G13 was isolated for the first time from the intestine of the mud crab through in vitro fermentation. The bacterium has probiotic properties and changes the gut microbiota to be beneficial to hosts in vitro as well as protecting hosts from Vibrio parahaemolyticus infection in vivo. The outcomes of this study indicate that C. butyricum G13 can be used as a potential probiotic in mud crab aquaculture.

Effects of dietary supplementation of Gracilaria lemaneiformis-derived sulfated polysaccharides on the growth, antioxidant capacity, and innate immunity of rabbitfish (Siganus canaliculatus).

The dietary supplementation of red seaweed-derived polysaccharides has been shown to be beneficial to fish and shellfish aquaculture. However, the function of red seaweed (Gracilaria lemaneiformis)-extracted polysaccharide (GLP) on the health status of rabbitfish (Siganus canaliculatus) is still unknown. This study explored the influences of GLP on growth performance, antioxidant activity, and immunity of rabbitfish. Herein, the fish were fed commercial pelleted feed incorporated with the diverse amount of GLP: 0 (control), 0.10 (GLP0.10), and 0.15 g kg-1 (GLP0.15) for 60 days. The results demonstrated that dietary GLP0.15 significantly elevated FBW and WG, while feed utilization efficiency improved (reduced feed conversion ratio and increased protein efficiency ratio) upon GLP0.10 treatment, regarding the control (P < 0.05). Also, dietary administration of GLP0.15 suggestively improved the serum acid phosphatase and lysozyme activity as well as hepatic total antioxidant capacity, catalase, and superoxide dismutase activity. In contrast, GLP0.15decreased the serum alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and malonaldehyde activity when compared to the control (P<0.05). Moreover, the lipase (36.08 and 16.46 U/mgprot in GLP0.10 and GLP0.15, respectively) and amylase (0.43 and 0.23 U/mgprot in GLP0.10 and GLP0.15, respectively) activity recorded the maximum values than the control (8.61 and 0.13 U/mgprot, respectively).Further, the intestinal morphometry was developed (such as increased villus length, width, and area) in the fish fed with a GLP-supplemented diet compared to the control. The KEGG pathway analysis unveiled that several differentially expressed genes (DEGs) in control vs. GLP0.10 and control vs. GLP0.15 were associated with metabolic or immune-associated pathways like antigen processing and presentation, phagosome, complement and coagulation cascades, and platelet activation. The DEGs, namely C3, f5, fgb, MHC1, and cfb, were evaluated in control vs. GLP0.10 and C3 and MHC1 in control vs. GLP0.15, suggesting their possible contributions to GLP-regulated immunity. Additionally, the cumulative mortality of rabbitfish after the Vibrio parahaemolyticus challenge was lower in both GLP0.10 (8.88%) and GLP0.15 (11.11%) than in control (33.33%) (P<0.05). Thus, these findings direct the potential use of GLP as an immunostimulant and growth promoter in rabbitfish aquaculture.

In vitro fermentation of Gracilaria lemaneiformis and its sulfated polysaccharides by rabbitfish gut microbes.

This study intended to characterize the Gracilaria lemaneiformis (SW)-derived polysaccharide (GLP) and explore the fermentation aspects of SW and GLP by rabbitfish (Siganus canaliculatus) intestinal microbes. The GLP was mainly composed of galactose and anhydrogalactose (at 2.0:0.75 molar ratio) with the linear mainstay of α-(1 â†’ 4) linked 3,6-anhydro-α-l-galactopyranose and ß-(1 â†’ 3)-linked galactopyranose units. The in vitro fermentation results showed that the SW and GLP could reinforce the short-chain fatty (SCFAs) production and change the diversity and composition of gut microbiota. Moreover, GLP boosted the Fusobacteria and reduced the Firmicutes abundance, while SW increased the Proteobacteria abundance. Furthermore, the adequacy of feasibly harmful bacteria (such as Vibrio) declined. Interestingly, most metabolic processes were correlated with the GLP and SW groups than the control and galactooligosaccharide (GOS)-treated groups. In addition, the intestinal microbes degrade the GLP with 88.21 % of the molecular weight reduction from 1.36 × 105 g/mol (at 0 h) to 1.6 × 104 g/mol (at 24 h). Therefore, the findings suggest that the SW and GLP have prebiotic potential and could be applied as functional feed additives in aquaculture.

Health benefits of butyrate and its producing bacterium, Clostridium butyricum, on aquatic animals.

Aquaculture plays an important role in contributing to global food security and nutrition; thus, the intensification and diversification of aquaculture are increasingly considered. However, paralleling the development of the industrial scale in aquaculture, the occurrence of diseases is always an important issue that causes great losses in economics. The finding of approaches that not only improve culture production but also reduce the impact of diseases in cultured animals is crucially essential. Previously, several studies have addressed the potential application of feed additives, such as prebiotics, probiotics, synbiotics, and microbial-derived metabolites (including short-chain fatty acids-SCFAs), in aquaculture. In this review, we provide an update focusing on the health benefits of dietary supplementation with a type of SCFAs, butyrate, and its producer, Clostridium butyricum, including their effects on growth, feed utilization, body composition, intestinal structure and function, antioxidant activity, immune response, and tolerance against stress and infection in aquatic animals. The outcomes of this study may indicate more benefits of the use of C. butyricum than that of butyrate (and its forms). This review provides general knowledge of the efficacy of butyrate and C. butyricum in aquaculture.

Role of Cellular Receptors in the Innate Immune System of Crustaceans in Response to White Spot Syndrome Virus.

Innate immunity is the only defense system for resistance against infections in crustaceans. In crustaceans, white spot diseases caused by white spot syndrome virus (WSSV) are a serious viral disease with high accumulative mortality after infection. Attachment and entry into cells have been known to be two initial and important steps in viral infection. However, systematic information about the mechanisms related to WSSV infection in crustaceans is still limited. Previous studies have reported that cellular receptors are important in the innate immune system and are responsible for the recognition of foreign microorganisms and in the stimulation of the immune responses during infections. In this review, we summarize the current understanding of the functions of cellular receptors, including Toll, C-type lectin, scavenger receptor, ß-integrin, polymeric immunoglobulin receptor, laminin receptor, globular C1q receptor, lipopolysaccharide-and ß-1,3-glucan-binding protein, chitin-binding protein, Ras-associated binding, and Down syndrome cell adhesion molecule in the innate immune defense of crustaceans, especially shrimp and crabs, in response to WSSV infection. The results of this study provide information on the interaction between viruses and hosts during infections, which is important in the development of preventative strategies and antiviral targets in cultured aquatic animals.

Serum hepatitis B viral (HBV) DNA is a predictive biomarker for survival in non-small cell lung cancer patients with chronic HBV infection.

Purpose: To study the association between pretreatment serum hepatitis B viral (HBV) DNA copy numbers and clinical outcome of non-small cell lung cancer (NSCLC) patients with chronic HBV infection. Patients and methods: We retrospectively evaluated the medical records of NSCLC HBV (+) patients between January 2008 and December 2010. The HBV DNA copy numbers and other prognostic factors including albumin (ALB), C-reactive protein (CRP), platelet, neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), and Glasgow Prognostic Score (GPS) were obtained before any antitumor treatment. Kaplan-Meier curves and the log-rank test were used to calculate prognostic significance. A multivariable Cox proportional hazard regression was modeled to analyze the independent prognostic factors for NSCLC HBV (+) patients. All independent prognostic factors in the Cox multivariable analysis were used to build a nomogram. The predictive accuracy of HBV DNA, TNM stage and nomogram was evaluated with the concordance index (C-index) and time-dependent receiver operating characteristics (ROC) curves, and simultaneously compared with traditional TNM staging system respectively. Results: A total of 188 patients were recruited in this study; the median age was 56 years, and the median overall survival (OS) was 34 months. Cox multivariate analysis results showed independent factors for OS including TNM stage (P=0.028), treatment (P=0.002), HBV DNA (P<0.001), and GPS (P=0.026). The nomogram model for survival was built based on four prognostic factors. The C-index for HBV DNA was 0.67, 0.69 for TNM stage, and 0.76 for the nomogram model. There was no statistical difference between HBV DNA and TNM stage (P=0.48). However, the C-index values of nomogram model were statistically higher both than HBV DNA (0.76 vs 0.67, P<0.001), and TNM stage (0.76 vs 0.69, P<0.001). Conclusion: Pretreatment serum HBV DNA copy numbers can act as a prognostic marker of survival for NSCLC patients with chronic HBV infection.

Fusobacterium nucleatum Aggravates the Progression of Colitis by Regulating M1 Macrophage Polarization via AKT2 Pathway.

Disordered intestinal flora and discordant immune response are associated with the development of ulcerative colitis (UC). Recent work has described the ability of macrophages to undergo repolarization toward a proinflammatory M1 or anti-inflammatory M2 phenotype in response to particular bacterium-derived signals. Fusobacterium nucleatum (F. nucleatum, Fn) is a species of intestinal commensal bacteria with potential pathogenicity, but its association with UC and how it may contribute to progression of UC is largely unknown. In this study, we provide new evidence that F. nucleatum accumulated heavily in the intestine of UC patients and was accompanied by the secretion of IFN-γ and the skewing of M1 macrophages. Mechanistically, our data showed that F. nucleatum aggravated dextran sodium sulfate (DSS)-induced colitis in the production of Th1-related cytokines IFN-γ through the AKT2 signaling pathway in vitro and in vivo. To further confirm the disease-relevance of these shifts in macrophage repolarization in response to F. nucleatum, stimulated bone marrow-derived macrophages (BMDMs) were transferred into recipient mice with DSS colitis. This transfer resulted in increased disease activity and inflammatory cytokine production. Taken together, we show clearly that F. nucleatum can promote the progression of UC via proinflammatory M1 macrophage skewing, and targeting F. nucleatum or AKT2 signaling may be a viable means of blocking development of UC.

A Fluorescent Coumarin-Based Probe for the Fast Detection of Cysteine with Live Cell Application.

A new coumarin-based fluorescent probe, containing an allylic esters group, has been designed and synthesized for sensing cysteine in physiological pH. In this fluorescent probe, the coumarin was applied as the fluorophore and an allylic esters group was combined as both a fluorescence quencher and a recognition unit. The probe can selectively and sensitively detect cysteine (Cys) over homocysteine, glutathione, and other amino acids, and has a rapid response time of 30 min and a low detection limit of 47.7 nM. In addition, the probe could be applied for cell imaging with low cytotoxicity.