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Effects of dietary Lactobacillus acidophilus and tussah immunoreactive substances (TIS) on disease resistance of sea cucumber (Apostichopus japonicus) against Vibrio splendidus were investigated. Sea cucumbers were fed with a basal diet in the control group (CK group) or were supplemented with L. acidophilus (L group), TIS (T group) or both (LT group). The challenge test with V. splendidus was conducted after the feeding trial. The cumulative mortality rate in the LT group was significantly lower, while the highest mortality occurred in the control group. The LT group showed better digestive and immune parameters compared with the other treatments. Dietary L. acidophilus and TIS in combination could effectively enhance digestive function and food uptake, depress oxidative damage and improve the organismal defence capability. Regarding gut microbiota profiles, the LT group was more capable of inhibiting potentially opportunistic pathogens and regulating intestinal flora balance compared with the other groups. The administration of L. acidophilus and TIS in the diet could effectively improve disease resistance against V. splendidus of sea cucumbers by improving the immune and digestive enzyme activities and modulating intestinal microflora, signalling a potential role in the development and production of aquatic feed. [ABSTRACT FROM AUTHOR]

Sea cucumber Apostichopus japonicus is an important economic species in echinoderms; however, its culture frequently suffered from diseases. Environmental stress was found to be a dominant inducer of A. japonicus diseases, and thus, discovering the characteristics of immune system under environmental stress is of great significance for disease control in A. japonicus culture. Phenoloxidase (PO) is recognized as a crucial immune‐related enzyme in invertebrates. In order to preliminarily illustrate the different roles that A. japonicus PO isoforms play under environmental stresses, the transcriptional expression of three identified A. japonicus PO isoforms (named as AjPOI, AjPOII and AjPOIII respectively) after different in vivo environmental stress simulations was determined with quantitative real‐time PCR method. Meanwhile, the transcriptional expression analysis of other representational immune‐related factors under environmental stresses was also performed for the evaluation of relationships between AjPOs and other immune‐related factors. The correlations between AjPOs and other immune‐related factors were analysed using Pearson correlation coefficient, and the effects of AjPOs on other immune‐related factors were then tested by dsRNA interference method. The results of transcriptional expression analysis indicated that AjPOI was involved in the resistance to low salinity, low temperature and hypoxia; AjPOII and AjPOIII were both required for the resistance to low salinity, high temperature, low temperature and hypoxia; and all three AjPOs were excluded from the response against high salinity. In addition, three AjPOs were found to take part in different periods of resistance to environmental stresses, and this property made A. japonicus PO system capable of outputting resistance activities continuously. The correlation analysis combined with dsRNA interference test revealed that AjPOI was capable of regulating AjCTSD, AjI‐LYZ and AjMTF expression; AjPOII and AjPOIII both played important roles in the regulation of AjC‐LYZ expression; and AjPOIII was also able to regulate AjToll expression. These findings suggested that A. japonicus PO isoforms function in innate immunity not only via catalysing the oxidative reactions, but also by broadly regulating the expression of other immune‐related factors. On the whole, three AjPOs showed obvious differences among each other in the resistance to environmental stresses and the regulation of other immune‐related factors, but comparatively, AjPOII and AjPOIII shared more similarity. [ABSTRACT FROM AUTHOR]

In order to understand the effects of seasonal change on the immunity of sea cucumber Apostichopus japonicus cultured in pond, A. japonicus with body weight of 12.2 ± 4.5 g (sample A) and 32.6 ± 7.1 g (sample B), respectively, were collected monthly and randomly from a typical pond during a year cycle and employed for the evaluation of immunocompetence. Simultaneously, the environmental factors in the pond including water temperature, pH, salinity and dissolved oxygen (DO) were measured using a handheld multiparameter meter. The activities of acid phosphatase (ACP), alkaline phosphatase (ALP), lysozyme ( LYZ), phenoloxidase (PO), superoxide dismutase ( SOD), catalase ( CAT) and myeloperoxidase ( MPO) in the coelomic fluid were selected for the evaluation of A. japonicus immunocompetence and determined using biochemical methods. The results showed that in both samples, the activities of all determined enzymes had small values in winter and early spring, and LYZ, CAT and MPO activities also presented small values in summer, suggesting that pond-cultured A. japonicus underwent immunosuppression twice during a year cycle, and the immunosuppression occurred in winter and early spring was more severe than that occurred in summer. In addition, most of the determined enzymes showed different variations between the two samples, and had significantly negative correlation with protein concentration, which was significantly and negatively correlated with water temperature, revealing that body weight and water temperature might have crucial effects on the immunity of A. japonicus cultured in pond. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the effects of Bacillus licheniformis and combined herbs extracts on the physiological and immune characteristics, intestinal microbiota, and vibriosis resistance of sea cucumber Apostichopus japonicus. The sea cucumbers were fed with basal diets supplemented with B. licheniformis (B group), combined herbs extracts (C group), or both (BC group). The BC group exhibited the better growth performance and higher digestive and immune enzyme activities, whereas the lowest parameters appeared in the control group. This revealed that dietary B. licheniformis and combined herbs extracts in combination could improve digestion, food intake, phosphatase-responsiveness, and oxidation resistance more efficiently compared with the other groups. The BC group was also more capable of regulating intestinal flora balance of sea cucumbers by increasing the beneficial bacteria related to energy synthesis and metabolic conversion, and inhibiting the potential pathogens associated with organic damage and metabolic disorders. Furthermore, the BC group showed greater ability of improving disease resistance by reducing the cumulative mortality rates and the counts of Vibrio splendidus. These results collectively demonstrated that dietary B. licheniformis and combined herbs extracts could improve the physiological and immune parameters, optimize the microbial community, and enhance the resistance to vibriosis of sea cucumbers, and the health-promoting effects were more obvious by supplying them in combination. [ABSTRACT FROM AUTHOR]

In recent years, sea cucumber has become a favorite healthcare food due to its characteristic prevention of cardiovascular diseases, suppression of tumors, as well as enhancement of immunity. In order to screen the anti-tumoral proteins or peptides from sea cucumber (Apostichopus japonicus), its cDNA library was analyzed, and a disintegrin-like and metalloproteinase with thrombospondin type 1 motif, member 13 (ADAMTS13)-like was found. ADAMTS13-like contains 10 thrombospondin 1 (TSP1) domains. Based on analysis of bioinformatics, the third TSP1 domain of this protein, which is further named Aj-Tspin, contains an arginine-glycine-aspartate (RGD) motif. Since our previous studies showed that the recombinant RGD-containing peptide from lampreys showed anti-tumoral activity, the third TSP1 domain of ADAMTS13-like was chosen to evaluate it's effect on tumor proliferation and metastasis, despite the fact it shares almost no homologue with disintegrins from other species. After artificial synthesis, its cDNA sequence, Aj-Tspin, which is composed of 56 amino acids, was subcloned into a pET23b vector and expressed as a recombinant Aj-Tspin (rAj-Tspin) in a soluble form with a molecular weight of 6.976 kDa. Through affinity chromatography, rAj-Tspin was purified as a single protein. Both anti-proliferation and immunofluorescence assays showed that rAj-Tspin suppressed the proliferation of Lewis lung carcinoma (LLC) cells through apoptosis. Adhesion assay also displayed that rAj-Tspin inhibited the adhesion of LLC cells to ECM proteins, including fibronectin, laminin, vitronectin and collagen. Lastly, rAj-Tspin also suppressed the migration and invasion of LLC cells across the filter in transwells. Thus, the above indicates that rAj-Tspin might act as a potential anti-tumoral drug in the future and could also provide information on the nutritional value of sea cucumber. [ABSTRACT FROM AUTHOR]