Abstract
We established an improved method for in vitro culture of glochidia in freshwater mussel Cristaria plicata and compared the transformation percentage in L-15 basic medium supplemented with different plasma from host fish or two non-host fish species. The results showed that the metamorphosis rate of glochidia cultured with host fish plasma and non-host fish plasma was all above 96%. High-density lipoprotein and low-density lipoprotein (LDL) levels were significantly lower in host fish plasma than in non-host fish plasma (P < 0.05), suggesting that LDL may be an important nutrient for glochidia metamorphosis. There were no significant differences in the 15-day survival and growth of juvenile mussels obtained from groups with non-host fish plasma under 1100/dish (C5) and a high concentration of antibiotics (D7), as compared with the control (A4) (P > 0.05). The method developed herein not only provides a simplified in vitro culture method for glochidia at a reduced cost, but it also improves the culture efficiency. The method and medium composition may be further improved in studies investigating the long-term growth of juvenile mussels, and it would greatly benefit artificial breeding and germplasm resource preservation for this important species.
Similar content being viewed by others
References
Bai, Y. Z., J. L. Li & B. B. Pan, 2008. Comparison of the parasitism effect of glochidia of triangle mussle (Hyriopsis cumingii) in five host fishes. Freshwater Fisheries 38(1): 3–5, 43.
Bieler, R., J. G. Carter & E. V. Coan, 2010. Classification of Bivalve Families. In Bouchet, P., & J.-P. Rocroi (eds), Nomenclator of Bivalve Families. Malacologia 52(2): 113–133.
Bogan, A. E., 2008. Global diversity of freshwater mussels (Mollusca, Bivalvia) in freshwater. Hydrobiologia 595(1): 139–147.
Christian, A. D. & J. L. Harris, 2010. An introduction to directions in freshwater mollusk conservation: molecules to ecosystems. Journal of the North American Benthological Society 27(2): 345–368.
Dodd, B. J., M. C. Barnhart & C. L. Rogers-Lowery, 2005. Cross-resistance of largemouth bass to glochidia of Unionid mussels. Journal of Parasitology 91(5): 1064–1072.
Gordon, M. E., J. B. Layzer & L. M. Madison, 1994. Glochidial host of Villosa taeniata (Mollusca: Unionoidea). Malacological Review 27: 113–114.
Haag, W. R. & M. L. Warren, 1999. Mantle displays of freshwater mussels elicit attacks from fish. Freshwater Biology 42(1): 35–40.
Hua, D., G. C. Xu, H. B. Wen & Y. F. Wang, 2005. Study on the host efficiency of the three kind of fish for Anodonta woodiana pacifica. Journal of Zhejiang Ocean University (Nature Science) 24(3): 213–216.
Hua D., R. J. Neves & Y. Jiao, 2013. Effects of algal density, water flow and substrate type on culturing juveniles of the rainbow mussel (Villosa iris) (Bivalvia: Unionidae) in a laboratory recirculating system [J]. Aquaculture, 195(1–2): 367–373.
Hudson, R. & C. Shelbourne, 1990. Improved in vitro culture of parasitic freshwater mussel glochidia. Report to the Tennessee Valley Authority, Muscle Shoals, AL.
Isom, B. G. & R. G. Hudson, 1982. In vitro culture of parasitic freshwater mussel glochidia. The Nautilus 96: 147–151.
Isom, B. G. & R. G. Hudson, 1984. Freshwater Mussels and Their Fish Hosts; physiological aspects. The Journal of Parasitology 70: 318–319.
Keller, A. E. & S. G. Zam, 1990. Simplification of in vitro culture techniques for freshwater mussels. Environmental Toxicology and Chemistry 9(10): 1291–1296.
Kovitvadhi, S., U. Kovitvadhi, P. Sawangwong, A. Thongpan, & J. Machado, 2006. Optimization of diet and culture environment for larvae and juvenile freshwater pearl mussels, Hyriopsis (Limnoscapha) myersiana Lea, 1856. Invertebrate Reproduction & Development 49(1): 61–70.
Lima, P., K. Uthaiwan, S. Kovitvadhi & J. Machado, 2006. In vitro culture of glochidia from the freshwater mussel Anodonta cygnea. Invertebrate Biology 125(1): 34–44.
Lima, P., M. L. Lima, K. Uthaiwan, K. Satit, C. Owen & J. Machado, 2012. A review on the “in vitro” culture of freshwater mussel (Unionoida). Hydrobiologia 691: 21–33.
Loveless, R., S. Raymond, J. Inman, R. Hudson & M. Fairfax, 1999. Isolation, identification and control of bacteria which threaten in vitro cultures of freshwater mussel glochidia. In Abstracts, 1st Symposium of the Freshwater Mollusk Conservation Society, Chattanooga, TN, March, p. 79.
Meyers, T. R., R. E. Millemann & C. A. Fustish, 1980. Glochidiosis of salmonid fishes. IV. Humoral and tissue responses of coho and Chinook salmon to experimental infection with Margaritifera margaritifera (L.) (Pelecypoda: Margaritanidae). Journal of Parasitology 66(2): 274–281.
Milam, C. D., J. L. Farris, J. Van-Hassel & M. L. Barfield, 2000. Reintroduction of native freshwater mussels using in vivo and in vitro propagation techniques. In Tankersley, R. A., D. I. Warmolts, G. T. Watters, B. J. Armitage, P. D. Johnson & R. S. Butler (eds.), Freshwater Mollusk Symposia Proceedings. Ohio Biological Survey, Columbus, OH: 53–61.
Owen, C. T., 2009. Investigations for the conservation and propagation of freshwater mussels. Dissertation, University of Louisville, Louisville.
Owen, C. T., J. E. Alexander & M. McGregor, 2010. Control of microbial contamination during in vitro culture of larval unionid mussels. Invertebrate Reproduction & Development 54(4): 187–193.
Reece, F. T., 2014. Studies on the In Vitro Propagation of Freshwater Mussels with Implications for Improving Juvenile Health. North Carolina State University, Raleigh, NC.
Reuling, F. H., 1919. Acquired immunity to an animal parasite. Journal of Infectious Diseases 24(1): 337–346.
Rogers-Lowery, C. L., R. V. Dimock Jr. & R. E. Kuhn, 2007. Antibody response of bluegill sunfish during development of acquired resistance against the larvae of the freshwater mussel Utterbackia imbecillis. Developmental and Comparative Immunology 31(2): 143–155.
Su, F. Y., H. J. Wang, B. Z. Pan, X. Q. Liu & H. Z. Wang, 2009. Assessment of species status of mollusca in the mid-lower Yangtze Lakes. Acta Hydrobiologica Sinica 33(6): 1051–1058.
Tankersley, R. A., 2000. Fluorescence techniques for evaluating lipid content of larval and juvenile mussels. In Tankersley, R. A., D. I. Warmolts, G. T. Watters, B. J. Armitage, P. D. Johnson & R. S. Butler (eds.), Freshwater Mollusk Symposia Proceedings. Ohio Biological Survey, Columbus, OH: 115–125.
Tedla, S & C. H. Fernando, 1969. Observations on the glochidia of Lampsilis radiata (Gmelin) infesting yellow perch, Perca flavescens (Mitchill) in the Bay of Quinte, Lake Ontario. Canadian Journal of Zoology 47: 705–712.
Uthaiwan, K., N. Noparatnaraporn & J. Machado, 2001. Culture of glochidia of the freshwater pearl mussel Hyriopsis myersiana (Lea, 1856) in artificial media. Aquaculture 195(195): 61–69.
Uthaiwan, K., P. Pakkong, N. Noparatnaraporn, L. Vilarinho & J. Machado, 2002. Study of a suitable fish plasma for in vitro culture of glochidia Hyriopsis myersiana. Aquaculture 209(1–4): 197–208.
Waller, D. L., L. E. Holland, L. G. Mitchell & T. W. Kammer, 1985. Artificial infestation of largemouth bass and walleye with glochidia of Lampsilis ventricosa (Pelecypoda: Unionidae). Freshwater Invertebrate Biology 4(3): 152–153.
Weaver, L. R., G. B. Pardue & R. J. Neves, 1991. Reproductive biology and fish hosts of the Tennessee Clubshell Pleurobema oviforme (Mollusca: Unionidae) in Virginia. American Midland Naturalist 126(1): 82–89.
Wen, H. B., 2009. Study of germplasm of major economic freshwater mollusks of China. Dissertation, Nanjing Agricultural University, Nanjing.
Wen, H. B., L. H. Qiu, R. B. Gu, G. C. Xu, D. Hua & P. Xu, 2009. The Host Efficiency of Two Kinds of Fish for Glochidia of Hyriopsis cumingii and Impact on Plasma Biochemical Indexes of Host Fish. Chinese Journal of Zoology 44(5): 92–97.
Wen, H. B., R. B. Gu, D. Hua, L. H. Qiu, G. C. Xu & P. Xu, 2011. Culture of glochidia of Hyriopsis cumingii in artificial media and morphological variation of transformed juvenile. Journal of Wuhan University (Nature Science Ed.) 57(1): 57–62.
Xie, M. Q. & G. Q. Li, 2003. Modern Parasites. Guangdong Science and Technology Press, Guangzhou.
Yeager, B. & C. F. Saylor, 1995. Fish hosts for four species of freshwater mussels (Pelecypoda: Unionidae) in the upper Tennessee River Drainage. American Midland Naturalist 133(1): 1–6.
Zhang, M. H., L. Xu, G. L. Xie, Y. B. Liu, X. M. Liu, S. C. Song, S. Ouyang & X. P. Wu, 2013. Species diversity, distribution and conservation of freshwater mollusk in Poyang Lake basin. Marine Sciences 37(8): 114–124.
Acknowledgments
The authors would like to thank Special Scientific Research Funds for Central Non-profit Institutes, Chinese Academy of Fishery Sciences (2016GH10), National Nonprofit Institute Research Grant of Freshwater Fisheries Research Center (2015JBFM02) and Ocean and Fishery Bureau of Jiangsu Province (Y2014-38), for providing financial support for this project. We also appreciate the insightful comments from the anonymous reviewers.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Guest editors: Manuel P. M. Lopes-Lima, Ronaldo G. Sousa, Lyuba E. Burlakova, Alexander Y. Karatayev & Knut Mehler / Ecology and Conservation of Freshwater Bivalves
Xue-Yan Ma and Hai-Bo Wen share the first authorship.
Rights and permissions
About this article
Cite this article
Ma, XY., Wen, HB., Zou, J. et al. An improved method for in vitro culture of glochidia in freshwater mussel Cristaria plicata (Mollusca, Bivalvia). Hydrobiologia 810, 133–144 (2018). https://doi.org/10.1007/s10750-016-3012-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10750-016-3012-3