Abstract
Germ cell cryopreservation has been used to preserve many fish species. However, this method has not been established for crustaceans; thus, we attempted to do this herein. The efficiency of slow freezing was compared to vitrification methods for germ cell cryopreservation in two types of marine shrimp, Fenneropenaeus merguiensis and Penaeus monodon. In situ hybridization with a vasa probe was used to identify germ cells. The effects of three cryoprotectants, dimethyl sulfoxide (DMSO), glycerol (GLY), and magnesium chloride (MgCl2), on germ cell viability and recovery rate were compared at three concentrations (5%, 10%, and 15%). The effects of thawing temperature, including 10 and 27 °C, were also investigated. We discovered that 10% DMSO with the vitrification is suitable for preserving the germ cells of F. merguiensis for a long time, whereas 10% GLY with vitrification is suitable for P. monodon. Moreover, the most suitable thawing temperature was 10 °C for both species. This is the first report of germ cell cryopreservation in crustaceans. Thus, we provide evidence that crustacean germ cells can be preserved long-term in liquid nitrogen; this is the first step in the sustainable preservation of crustaceans, especially shrimp.
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References
Akarasanon K, Damrongphol P, Poolsanguan W (2004) Long-term cryopreservation of spermatophore of the giant freshwater prawn, Macrobrachium rosenbergii (de Man). Aquac Res 35:1415–1420
Castelo-Branco T, Batista AM, Guerra MMP, Soares R, Peixoto S (2015) Sperm vitrification in the white shrimp Litopenaeus vannamei. Aquaculture 436:110–113
Chaves TCB, Fernandes AB, Mello MRB, Oshiro LMY (2014) Cryopreservation and sperm storage of the white shrimp. Bol Do Inst Pesca 40:17–22
Fahy GM, MacFarlane DR, Angell CA, Meryman HT (1984) Vitrification as an approach to cryopreservation. Cryobiology 21:407–426
FAO (2020) The state of world fisheries and aquaculture 2020. Sustainability in Action. Rome. https://doi.org/10.4060/ca9229en
Fatihah SN, Jasmani S, Abol-Munafi AB, Noorbaiduri S, Muhd-Farouk H, Ikhwanuddin M (2016) Development of a sperm cryopreservation protocol for the mud spiny lobster, Panulirus polyphagus. Aquaculture 426:56–63
Hayashi M, Ichida K, Sadaie S, Miwa M, Fujihara R, Nagasaka Y, Yoshizaki G (2019) Establishment of novel monoclonal antibodies for identification of type A spermatogonia in teleosts. Biol Reprod 101:478–491
Ichida K, Kawamura W, Miwa M, Iwasaki Y, Kubokawa T, Hayashi M, Yazawa R, Yoshizaki G (2019) Specific visualization of live type A spermatogonia of Pacific bluefin tuna using fluorescent dye-conjugated antibodies. Biol Reprod 100:1637–1647
Kang X, Li G, Mu S, Guo M, Ge S (2009) Acrosome reaction of Chinese mitten-handed crab Eriocheir sinensis (Crustacea: Decapoda) spermatozoa: promoted by long-term cryopreservation. Aquaculture 295:195–199
Karlsson JOM (2001) A theoretical model of intracellular devitrification. Cryobiology 42:154–169
Kobayashi T, Takeuchi Y, Yoshizaki G, Takeuchi T (2003) Cryopreservation of trout primordial germ cells. Fish Physiol Biochem 28:479–480
Lee S, Iwasaki Y, Shikina S, Yoshizaki G (2013) Generation of functional eggs and sperm from cryopreserved whole testes. Proc Natl Acad Sci USA 110:1640–1645
Lee S, Iwasaki Y, Yoshizaki G (2016) Cryobiology Long-term (5 years) cryopreserved spermatogonia have high capacity to generate functional gametes via interspecies transplantation in salmonids. Cryobiology 73:286–290
Memon AJ, Talpur AD, Khan MI, Fanddudm MO, Safiah J, Abol-Munafi AB, Ikhwanuddin M (2012) Optimization of spermatophores cryopreservation protocol of banana shrimp (Penaeus merguiensis) (De Man, 1888). J Anim Vet Adv 11:1688–1704
Morita T, Miwa M, Kumakura N, Morishima K, Miki T, Takeuch Y, Yoshizaki G (2021) Production of functional sperm from cryopreserved testicular germ cells following intraperitoneal transplantation into allogeneic surrogate in yellowtail (Seriola quinqueradiata). Cryobiology 100:32–39
Nagasawa K, Takeuchi Y, Miwa M, Higuchi K, Morita T, Mitsuboshi T, Miyaki K, Kadomura K, Yoshizaki G (2009) cDNA cloning and expression analysis of a vasa-like gene in Pacific bluefin tuna Thunnus orientalis. Fish Sci 75:71–79
Narasimman S, Krishnamoorthy D, Buelah CD, Natesan M (2018) Retrieval and cryopreservation of sperm in spermatophores from cadaveric Indian white shrimp, Fenneropenaeus indicus (H. Milne Edwards, 1837). Anim Reprod Sci 192:185–192
Nimrat S, Noppakun S, Sripuak K, Boonthai T, Vuthiphandchai V (2020) Cryopreservation of banana shrimp (Fenneropenaeus merguiensis) spermatophores with supplementation of medicinal plant extracts: development of a programmable controlled-rate method and a practical method. Aquaculture 515:734537
Octavera A, Yoshizaki G (2020) Production of Chinese rosy bitterling offspring derived from frozen and vitrified whole testis by spermatogonial transplantation. Fish Physiol Biochem 46:1431–1442
Pegg DE (2015) Principles of cryopreservation. In: Wolkers W, Oldenhof H (eds) Cryopreservation and freeze-drying protocols, Springer, New York, NY, pp 3–19
Pšenička M, Saito T, Rodina M, Dzyuba B (2016) Cryopreservation of early stage Siberian sturgeon Acipenser baerii germ cells, comparison of whole tissue and dissociated cells. Cryobiology 72:119–122
Seki S, Kusano K, Lee S, Iwasaki Y, Masaru Y (2017) Production of the medaka derived from vitrified whole testes by germ cell transplantation. Sci Rep 7:43185
Vuthiphandchai V, Nimrat S, Kotcharat S, Bart AN (2007) Development of a cryopreservation protocol for long-term storage of black tiger shrimp (Penaeus monodon) spermatophores. Theriogenology 68:1192–1199
Wowk B, Leitl E, Rasch CM, Mesbah-Karimi N, Harris SB, Fahy GM (2000) Vitrification enhancement by synthetic ice blocking agents. Cryobiology 40:228–236
Yano A, Suzuki K, Yoshizaki G (2008) Flow-cytometric isolation of testicular germ cells from rainbow trout (Oncorhynchus mykiss) carrying the green fluorescent protein gene driven by trout vasa regulatory regions. Biol Reprod 78:151–158
Ye H, Zhou C, Yue H, Wu M, Ruan R, Du H, Li C, Wei Q (2021) Cryopreservation of germline stem cells in American paddlefish (Polyodon spathula). Anim Reprod Sci 224:106667
Yoshikawa H, Ino Y, Shigenaga K, Katayama T (2018) Production of tiger puffer Takifugu rubripes from cryopreserved testicular germ cells using surrogate broodstock technology. Aquaculture 493:302–313
Acknowledgements
We thank Dr. G. Yoshizaki and R. Yazawa from Tokyo University of Marine Science and Technology for the valuable discussion and technical support during the study. We thank the Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University for the supporting equipment in this study.
Funding
This work was supported by grants from Thailand National Research Fund, Prince of Songkla University, under the project: Preservation of banana shrimp for sustainable aquaculture and the environment, grant No. SCI6201034S, and the Japan Science and Technology Agency/Japan International Cooperation Agency, Science and Technology Research Partnership for Sustainable Development, SATREPS (JPMJSA1806), Project for Utilization of Thailand Local Genetic Resources to Develop Novel Farmed Fish for Global Market (Thai Fish Project).
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Rakbanjong, N., Okutsu, T., Chotigeat, W. et al. Cryopreservation of Germ Cells of Banana Shrimp (Fenneropenaeus merguiensis) and Black Tiger Shrimp (Penaeus monodon). Mar Biotechnol 23, 590–601 (2021). https://doi.org/10.1007/s10126-021-10048-1
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DOI: https://doi.org/10.1007/s10126-021-10048-1