High-Density Cultivation of the Marine Ciliate Uronema marinum (Ciliophora, Oligohymenophorea) in Axenic Medium

Weibo Zheng,

Feng Gao,

Alan Warren


Uronema marinum is a cosmopolitan marine ciliate. It is a facultative parasite and the main causative agent of outbreaks of scuticociliatosis in aquaculture fish. This study reports a method for the axenic cultivation of U. marinum in high densities in an artificial medium comprising proteose peptone, glucose and yeast extract powder as its basic components. The absence of bacteria in the cultures was confirmed by fluorescence microscopy of DAPI-stained samples and the failure to recover bacterial SSU-rDNA using standard PCR methods. Using this axenic medium, a maximum cell density of 420,000 ciliate cells/ml was achieved, which is significantly higher than in cultures using living bacteria as food or in other axenic media reported previously. This method for high-density axenic cultivation of U. marinum should facilitate future research on this economically important facultative fish parasite.

Słowa kluczowe: Axenic cultivation, ciliates, fish parasite, scuticociliatosis, Uronema marinum

Anderson S. A., Hulston D. A., McVeagh S. M., Webb V. L., Smith P. J. (2009) In vitro culture and cryopreservation of Uronema marinum isolated from farmed New Zealand groper (Polyprion oxygeneios). J. Microbiol. Meth. 79: 62–66

Azad I. S., Al-Marzouk A., James C. M., Almatar S., Al-Gharabally H. (2007) Scuticociliatosis-associated mortalities and histopathology of natural infection in cultured silver pomfret (Pampus argenteus Euphrasen) in Kuwait. Aquaculture 262: 202–210

Corliss J. O. (1952) Comparative studies on holotrichous ciliates in the Colpidium-Glaucoma-Leucophrys-Tetrahymena group. 1. General considerations and history of strains in pure culture Trans. Amer. Microscop. Soc. 71: 159–184

Curds C. R., Cockburn A. (1968) Studies on the growth and feeding of Tetrahymena pyriformis in axenic and monoxenic culture. J. Gen. Mircrobiol. 54: 343–358

Foissner W., Wilbert N. (1981) A comparative study of the infraciliature and silverline system of the fresh-water scuticociliates Pseudocohnilembus putrinus (Kahl, 1928) nov. comb., P. pusillus (Quennerstedt, 1869) nov. comb., and the marine form P. marinus Thompson, 1966. J. Protozool. 28: 291–297

Gast V. (1985) Bacteria as a food source for microzooplankton in the Schlei Fjord and Baltic Sea with special reference to ciliates. Mar. Ecol. Prog. Ser. 22: 107–120

Glaser R. W., Coria N. A. (1933) The culture of Paramecium caudatum free from living microorganisms. J. Parasit. 20: 33–37

Hanna B. A., Lilly D. M. (1970) Axenic cultivation of Uronema marinumAm. Zool. 10: 539–540

Hanna B. A., Lilly D. M. (1974) Growth of Uronema marinum in chemically defined medium. Mar. Biol. 26: 153–160

Jee B., Kim Y., Park M. S. (2001) Morphology and biology of parasite responsible for scuticociliatosis of cultured olive flounder Paralichthys olivaceusDis. Aquat. Organ. 47: 49–55

Jin C., Harikrishnan R., Moon Y., Kim M., Kim J., Balasundaram C., Azad I. S., Heo M. (2009) Histopathological changes of Korea cultured olive flounder, Paralichthys olivaceus due to scuticociliatosis caused by histophagous scuticociliate, Philasterides dicentrarachiVet. Parasitol. 161: 292–301

Kim H., Choi S., Choi S., Kim M., Kim S., Nam Y., Kim K. (2004) Effects of bile acids on proliferation and production of proteinase activity of Uronema marinum (Ciliophora: Scuticociliatida). Dis. Aquat. Organ. 62: 245–248

Kwon S., Kim C., Kim K. (2003) Differences between short- and long-term cultures of Uronema marinum (Ciliophora: Scuticociliatida) in chemiluminescence inhibitory activity, antioxidative enzyme and protease activity. Aquaculture 221: 107–114

Lee E., Kim C., Cho J., Ahn K., Kim K. (2003) Measurement of protease activity of live Uronema marinum (Ciliata: Scuticociliatida) by fluorescence polarization. Dis. Aquat. Organ. 54: 85–88

Martin C. E., Hiramitsu K., Kitajima Y., Nozawa Y., Skriver L., Thompson Jr. G. A. (1976) Molecular control of membrane properties during temperature acclimation. Fatty acid desaturase regulation of membrane fluidity in acclimating Tetrahymena cells. Biochemistry 15: 5218–5227

Osborne C. A., Galic M., Sangwan P., Janssen P. H. (2005) PCRgenerated artefact from 16S rRNA gene-specific primers. Fems Microbiol. Lett. 248: 183–187

Pan H., Huang J., Hu X., Fan X., Al-Rasheid K. A. S., Song W. (2010) Morphology and SSU rRNA gene sequences of three marine ciliates from Yellow Sea, China, including one new species, Uronema heteromarinum nov. spec. (Ciliophora, Scuticociliatida). Acta Protozool. 49: 45–59

Parker J. G. (1979) Toxic effects of heavy metals upon cultures of Uronema marinum (Ciliophora: Uronematidae). Mar. Biol. 54: 17–24

Soldo A. T., Merlin E. J. (1972) The cultivation of symbiote-free marine ciliates in axenic medium. J. Protozool. 19: 519–524

Song J., Kitamura S., Oh M., Kang H., Lee J., Tanaka S., Jung S. (2009) Pathogenicity of Miamiensis avidus (syn. Philasterides dicentrarchi), Pseudocohnilembus persalinusPseudocohnilembus hargisi and Uronema marinum (Ciliophora, Scuticociliatida). Dis. Aquat. Organ. 83: 133–143

Thiele J., Honer-Schmid O., Wahl J., Kleefeld G., Schultz J. (1980) A new method for axenic mass cultivation of Paramecium tetraureliaJ. Protozool. 27: 118–121

Wang Y., Song W., Warren A., Al-Rasheid K. A. S., Al-Quraishy S. A., Al-Farraj S. A., Hu X., Pan H. (2009) Descriptions of two new marine scuticociliates, Pleuronema sinica n. sp. and P. wilberti n. sp. (Ciliophora: Scuticociliatida), from the Yellow Sea, China. Eur. J. Protistol. 45: 29–37

Czasopismo w darmowym dostępie.

Czasopismo ukazuje się w sposób ciągły on-line.
Pierwotną formą czasopisma jest wersja elektroniczna.