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Lentinan extracted from shiitake mushrooms (Lentinus edodes) improves the non-specific immunity of sea cucumber (Apostichopus japonicus)

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Abstract

In this study, the subpopulations of coelomocytes of sea cucumber Apostichopus japonicus were identified with differential-interference microscope and flow cytometry (FCM). The immunostimulatory potential of crude lentinan (cLNT) from shiitake mushrooms (Lentinus edodes) on A. japonicus was then investigated both in vitro and in vivo. In vitro, the coelomocytes of A. japonicus were incubated with different concentrations of cLNT (0, 2.5, 7.5, 22.5 mg/L) for 1 h, and immuno assays were performed. Flow cytometric analysis revealed that phagocytic activity and viability of coelomocytes increased significantly with cLNT treatment. Biochemical analysis showed a significant increase in peroxidase and alkaline phosphatase activity in coelomocytes. In vivo, the immunostimulatory effect of cLNT on coelomocytes was investigated with FCM by feeding A. japonicus with cLNT-supplemented or control diet for 24 days. The phagocytic activity of coelomocytes in cLNT treatment group was significantly increased at 4, 8 and 12 days of feeding. The viability of coelomocytes was increased after 8 days of feeding. At 4, 8, 12 and 18 days of feeding, the number of small-sized and low granularity coelomocytes was reduced, while the number of large-sized and high granularity coelomocytes was increased compared to control diet. Collectively, our in vitro and in vivo studies suggest that cLNT derived from L. edodes clearly reveals immunostimulating effects on A. japonicus by increasing the viability and phagocytic activity of coelomocytes, and these findings suggest that dietary supplementation of cLNT might further improve the commercial production of sea cucumbers.

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Abbreviations

ACP:

Acid phosphatase

AKP:

Alkaline phosphatase

cLNT:

Crude lentinan

CF:

Coelomic fluid containing coelomocytes

CLS:

Coelomocytes lysate supernatant

FCM:

Flow cytometry

FSC:

Forward-scatter parameters

LPS:

Lipopolysaccharides

PI:

Propidium iodide

POD:

Peroxidase

SSC:

Side-scatter parameters

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Acknowledgments

This work was supported by a fund from the Shandong Province Natural Science Foundation of China, NO. ZR2010CM017 and NO. ZR2012CM004. Thanks go to Wendy Lee and Qi Sun for editorial review of the manuscript.

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Authors and Affiliations

  1. College of Life Science, Ludong University, 186 Hongqi Zhonglu, Yantai, 264025, Shandong, People’s Republic of China

    Yiyan Wang, Haina Wang, Dongchun Yan, Lei Wang, Zhenxing Sun & Hushan Sun

  2. Weihai Tashan High School, Weihai, Shandong, People’s Republic of China

    Haina Wang

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  1. Yiyan Wang
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  2. Haina Wang
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  4. Lei Wang
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Correspondence to Hushan Sun.

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Wang, Y., Wang, H., Yan, D. et al. Lentinan extracted from shiitake mushrooms (Lentinus edodes) improves the non-specific immunity of sea cucumber (Apostichopus japonicus). Aquacult Int 21, 1261–1277 (2013). https://doi.org/10.1007/s10499-013-9628-7

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