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Preliminary Characterization of Hemocyte and Immunity of Asian Horseshoe Crabs, Tachypleus tridentatus, and Carcinoscorpius rotundicauda in Captivity

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International Horseshoe Crab Conservation and Research Efforts: 2007- 2020

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Abstract

Overfishing for bleeding has been identified as the major attribute to the substantial decline of Asian horseshoe crab populations. Owing to the low willingness of Tachypleus amebocyte lysate (TAL) firms to return bled horseshoe crabs to the sea, the pond culture practice has been proposed as the alternative to keep wild-caught adults or subadults in outdoor ponds for years for repeated bleeding. The potential effects of the prolonged culture on hemolymph quality of horseshoe crabs are questioned since their general health status has an impact on the TAL production quality and the resource sustainability for bleeding. However, the baseline information regarding immune-related parameters of horseshoe crabs, particularly the Asian species, is very limited. A lacking of standardized classification scheme of horseshoe crab hemocytes makes the health indicator measurement, such as differential hemocyte counts, and the comparison among studies more challenging. In this study, hemocyte morphology and immune-related enzymatic activity in T. tridentatus and Carcinoscorpius rotundicauda were determined, in which the individuals had been kept in outdoor ponds for one year. Wright–Giemsa staining result demonstrated that three hemocyte types, namely granulocyte, non-granulocyte, and macrophage, were found. Granulocyte was the major cell type in both species, and the other two hemocyte types were noted in very small proportions surrounded by the basophil granulocytes. The cytoplasmic granules in granulocytes were stained in either blue (basophil) or purple–red (eosinophil), but the macrophages contain a mixture of blue- and red-stained granules. For alkaline phosphatase (AKP) and glutathione peroxidase (GSH-Px) activities, there were no significant differences either between species or among digestive tract tissues, except that a statistical higher AKP activity level was noted in T. tridentatus serum than in Carcinoscorpius rotundicauda. The AKP and GSH-Px activity levels, in general, were higher compared to those reported in smaller-sized aquatic invertebrates. The findings may be useful as the baseline data in further immunological studies of Asian horseshoe crabs, especially for developing health monitoring system and improving sustainable bleeding practices.

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Acknowledgements

This work was supported by “Special Fund for Basic Scientific Research of Central Public Research Institutes” of the South China Sea Fisheries Research Institute, Chinese Academy of Fishery (2019TS21), Guangxi BaGui Youth Scholars Programme, and Guangxi Recruitment Program of 100 Global Experts.

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Authors and Affiliations

  1. Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, South China Sea Fisheries Research Institute, Chinese Academy of Fishery, Guangzhou, China

    Xiaoyong Xie, Mujiao Xie, Guoling Ye & Yuyuan Bao

  2. Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, College of Marine Sciences, Beibu Gulf University, Qinzhou, China

    Kit Yue Kwan

  3. Guangdong Marine Development and Planning Research Centre, Guangzhou, China

    Jinxiang Zhong

  4. Guangdong Marine Development and Planning Research Centre, Guangzhou, China

    Yuyuan Bao

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  1. Xiaoyong Xie
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  2. Kit Yue Kwan
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  3. Jinxiang Zhong
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  4. Mujiao Xie
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  5. Guoling Ye
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Correspondence to Kit Yue Kwan .

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Editors and Affiliations

  1. Department of Biology, Chemistry, and Environmental Studies (BCES) Molloy College, Center for Environmental Research and Coastal Oceans Monitoring (CERCOM), Earth and Environmental Sciences, West Sayville, NY, USA

    John T. Tanacredi

  2. Department of Natural Sciences, Fordham University, New York, NY, USA

    Mark L. Botton

  3. Department of Chemistry, City University of Hong Kong, Hong Kong, China

    Paul K. S. Shin

  4. Center for Environmental Research and Coastal Oceans Monitoring (CERCOM), Molloy College, West Sayville, NY, USA

    Yumiko Iwasaki

  5. Department of Chemistry, City University of Hong Kong, Hong Kong, China

    Siu Gin Cheung

  6. Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, Beibu Gulf University, Qinzhou, China

    Kit Yue Kwan

  7. Department of Biology, Sacred Heart University, Fairfield, CT, USA

    Jennifer H. Mattei

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Xie, X., Kwan, K.Y., Zhong, J., Xie, M., Ye, G., Bao, Y. (2022). Preliminary Characterization of Hemocyte and Immunity of Asian Horseshoe Crabs, Tachypleus tridentatus, and Carcinoscorpius rotundicauda in Captivity. In: Tanacredi, J.T., et al. International Horseshoe Crab Conservation and Research Efforts: 2007- 2020. Springer, Cham. https://doi.org/10.1007/978-3-030-82315-3_11

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