Skip to main content
SpringerLink
Log in

Development of two brain cell lines from goldfish and silver crucian carp and viral susceptibility to Cyprinid herpesivirus-2

  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

Two cell lines were established from silver crucian carp and goldfish brain tissue and used as the biological tool for monitoring viral diseases. Characterization including optimal growth kinetics study, karyotyping, and mitochondrial ribosomal RNA (rRNA) genotyping were performed. The primary cultures of these cells were generated by the explant technique using the medium 199 supplemented with 20 % fetal bovine serum and epidermal/fibroblast growth factors. The cells grew over the range of 15 to 30°C, while the optimal temperature for culture was 30°C. The cell lines were maintained in vitro and could be subcultured over 40 times. Following cryopreservation in liquid nitrogen, thawed cells exhibit viability of > 90 % after a 13-mo period of storage. The chromosome count of two cell lines were determined to be 154 and 110, respectively, which agreed well with triploid crucian carp brain cells and diploid goldfish brain cells. Polymerase chain reaction amplification and sequence analysis indicated 100 % and 94% match with known crucian carp mitochondrial DNA sequences. Cytopathic effect was continuously observed in both cell lines over 10 passages after inoculation with tissue homogenates of sick or died goldfish from cyprinid herpesvirus 2 (CyHV-2) outbreaks. These newly established cell lines could be a diagnostic tool for viral diseases in fish species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 6.

Similar content being viewed by others

References

  • Apalikova OV, Podlesnykh AV, Kukhlevsky AD, Guohua S, Brykov VA (2011) Phylogenetic relationships of silver crusian carp Carassius auratus gibelio, C. auratus cuvieri, crucian carp Carassius carassius, and common carp Cyprinus carpio as inferred from mitochondrial DNA variation. Russ J Genet 47(3):322–331

    Article  CAS  Google Scholar 

  • Chang PH, Lee SH, Chiang HC, Jong MH (1999) Epizootic of herpes-like virus infection in goldfish, Carassius auratus, in Taiwan. Fish Pathol 34(4):209–210

    Article  Google Scholar 

  • Chen SL, Ren GC, Sha ZX, Shi CY (2004) Establishment of a continuous embryonic cell line from Japanese flounder Pararlichthys olivaceus for virus isolation. Dis Aquat Org 60:241–246

    Article  Google Scholar 

  • Chen SL, Sha ZX, Ye HQ (2003a) Establishment of a pluripotent embryonic cell line from sea perch blastula embryo. Aquaculture 218:141–151

    Article  CAS  Google Scholar 

  • Chen SL, Ye HQ, Sha ZX, Hong Y (2003b) Derivation of a pluripotent embryonic cell line from red sea bream blastulas. J Fish Biol 63:795–805

    Article  Google Scholar 

  • DanÄ›k T, Kalous L, Vesel T, Krásová E, Reschová S, Rylková K, Kulich P, Petrtýl M, Pokorová D, Knytl M (2012) Massive mortality of Prussian carp Carassius gibelio in the upper Elbe basin associated with herpesviral hematopoietic necrosis (CyHV-2). Dis Aquat Org 102(2):87–95

    Article  Google Scholar 

  • Davison AJ, Eberle R, Ehlers B, Hayward GS, McGeoch DJ, Minson AC, Pellett PE, Roizman B, Studdert MJ, Thiry E (2009) The order Herpesvirales. Arch Virol 154(1):171–177

    Article  CAS  Google Scholar 

  • Ding Z, Xia S, Zhao Z, Xia A, Shen M, Tang J, Xue H, Geng X, Yuan S (2014) Histopathological characterization and fluorescence in situ hybridization of Cyprinid herpesvirus 2 in cultured Prussian carp, Carassius auratus gibelio in China. J Virol Methods 206:76–83

    Article  CAS  Google Scholar 

  • FAO (2014) Fishery and Aquaculture Statistics. Global aquaculture production. 1950-2014(Fishstat). In: FAO FiSheries and Aquaculture Department [online]. Rome. Updated 2018. www.fao.org/fishery/statistics/software/fishstatj/en

  • FAO (2018) Cultured Aquatic Species Information Programme. Carassius carassius. Text by Weimin M. In: FAO Fisheries and Aquaculture Department. Rome

  • Fichi G, Cardeti G, Cocumelli C, Vendramin N, Toffan A, Eleni C, Siemoni N, Fischetti R, Susini F (2013) Detection of Cyprinid herpesvirus 2 in association with an Aeromonas sobria infection of Carassius carassius (L.), in Italy. J Fish Dis 36(10):823–830

    CAS  Google Scholar 

  • Freshney RI ed (1994). Culture of animal cells, A Manual of Basic Techniques New York: pp 486

  • Giovannini S, Bergmann SM, Keeling C, Lany C, Schütze H, Schmidt-Posthaus H (2016) Herpesviral hematopoietic necrosis in goldfish in Switzerland: early lesions in clinically normal goldfish (Carassius auratus). Vet Pathol 53(4):847–852

    Article  CAS  Google Scholar 

  • Goodwin AE, Khoo L, LaPatra SE, Bonar A, Key DW, Garner M, V. Lee, M & Hanson, Larry. (2006) Goldfish hematopoietic necrosis herpesvirus (cyprinid herpesvirus 2) in the USA: molecular confirmation of isolates from diseased fish. J Aquat Anim Health 18(1): 11–18

  • Groff JM, LaPatra SE, Munn RJ, Zinkl JG (1998) A viral epizootic in cultured populations of juvenile goldfish due to a putative herpesvirus etiology. J Vet Diagn Investig 10(4):375–378

    Article  CAS  Google Scholar 

  • Hanson L, Dishon A, Kotler M (2011) Herpesviruses that infect fish. Viruses 3(11):216–219

    Article  Google Scholar 

  • Hedrick RP, Waltzek TB, McDowell TS (2006) Susceptibility of koi carp, common carp, goldfish, and goldfish x common carp hybrids to cyprinid herpesvirus-2 and herpesvirus-3. J Aquat Anim Health 18(1):26–34

    Article  Google Scholar 

  • Hong YJ, Yu ZJ, Zhou L, Gui JF (2005) A population of red-transparent, triploid Carassius auratus. J Fish Biol 67(4):1139–1143

    Article  Google Scholar 

  • Ito T, Kurita J, Ozaki A, Sano M, Fukuda H, Ototake M (2013) Growth of cyprinid herpesvirus 2 (CyHV-2) in cell culture and experimental infection of goldfish Carassius auratus. Dis Aquat Org 105(3):192–202

    Article  Google Scholar 

  • Ito T, Maeno Y (2014) Susceptibility of Japanese Cyprininae fish species to cyprinid herpesvirus 2 (CyHV-2). Vet Microbiol 169(3–4):128–134

    Article  CAS  Google Scholar 

  • Jeffery KR, Bateman K, Bayley A, Feist SW, Hulland J, Longshaw C, Stone D, Woolford G, Way K (2007) Isolation of a cyprinid herpesvirus 2 from goldfish, Carassius auratus (L.), in the UK. J Fish Dis 30(11):649–656

    Article  CAS  Google Scholar 

  • Jung SJ, Miyazaki T (1995) Herpesviral haematopoietic necrosis of goldfish, Carassius auratus (L.). J. Fish Dis 18(3):211–220

    Article  Google Scholar 

  • Komiyama T, Kobayashi H, Tateno Y, Inoko H, Gojobori T, Ikeo K (2009) An evolutionary origin and selection process of goldfish. Gene 430(1–2): 0–11

  • Lakra WS, Swaminathan TR, Joy KP (2011) Development, characterization, conservation and storage of fish cell lines: a review. Fish Physiol Biochem 37(1):1–20

    Article  CAS  Google Scholar 

  • Lovy J, Friend SE (2014) Cyprinid herpesvirus-2 causing mass mortality in goldfish: applying electron microscopy to histological samples for diagnostic virology. Dis Aquat Org 108(1):1–9

    Article  CAS  Google Scholar 

  • Ma J, Zhou Y, Fan Y, Liu W, Jiang N, Zeng L (2016) The physical-chemical and biological characteristics of Cyprinid herpesvirus 2 and its ultrastructural morphogenesis in vitro. J Fish China 40(3):475–483

    Google Scholar 

  • Ohno S, Muramoto J, Christian L, Atkin NB (1967) Diploid-tetraploid relationship among old-world members of the fish family Cyprinidae. Chromosom 23:1–9

    Article  Google Scholar 

  • Radosavljević V, Maksimović ZJ, Veljović L, Ljubojević D, Ćirković M, Marković Z, Vesna M (2017) Cyprinid herpesvirus diseases. Arch Vet Med 10(1):51–60

    Google Scholar 

  • Rougée L, Ostrander RH, Richmond YL (2007) Establishment, characterization, and viral susceptibility of two cell lines derived from goldfish Carassius auratus muscle and swim bladder. Dis Aquat Org 77:127–135

    Article  Google Scholar 

  • Sahoo PK, Swaminathan TR, Abraham TJ, Kumar R, Pattanayak S, Mohapatra A, Rath SS, Patra A, Adikesavalu H, Sood N, Pradhan PK, Das BK, Jayasankar P, Jena JK (2016) Detection of goldfish haematopoietic necrosis herpes virus (Cyprinid herpesvirus-2) with multi-drug resistant Aeromonas hydrophila infection in goldfish: first evidence of any viral disease outbreak in ornamental freshwater aquaculture farms in India. Acta Trop 161:8–17

    Article  CAS  Google Scholar 

  • Shibata T, Nanjo A, Saito M, Yoshii K, Ito T, Nakanishi T, Sakamoto T, Sano M (2015) In vitro characteristics of cyprinid herpesvirus 2: effect of kidney extract supplementation on growth. Dis Aquat Org 115(3):223–232

    Article  Google Scholar 

  • Stephens FJ, Raidal SR, Jones B (2004) Haematopoietic necrosis in a goldfish (Carassius auratus) associated with an agent morphologically similar to herpesvirus. Aust Vet J 82(3):167–169

    Article  CAS  Google Scholar 

  • Wang L, He J, Liang L, Zheng X, Jia P, Shi XJ, Lan W, Xie J, Liu Hongxi XP (2012) Mass mortality caused by Cyprinid Herpes virus 2(CyHV-2)in Prussian carp (Carassius gibelio) in China. Bull Eur Assoc Fish Pathol 32(5):164–173

    Google Scholar 

  • Xia S, Wang H, Podok P, Xu D, Jiang Y, Lü L, Sano M (2016) The expression analysis of immune genes and microscopic morphology of CCF cells in response to Cyprinid herpesvirus 2 infection. J Fish China 40(12):1915–1922

    Google Scholar 

  • Xu J, Zeng L, Zhang H, Zhou Y, Ma J, Fan Y (2013) Cyprinid herpes virus 2 infection emerged in cultured gibel carp, Carassius auratus gibelio in China. Vet Microbiol 166(12):138–144

    Article  Google Scholar 

  • Yi Z, Biqian L, Jie T (2012) Identifying six strains of Carassius auratus by PCR-RFLP on mtDNA ND3/ND4L/ND4 gene. Biotechnol Bull 11:144–149

    Article  Google Scholar 

Download references

Funding

This work was financially supported by the science and technology project of Jiangsu Entry-Exit Inspection and Quarantine Bureau of P.R. China (No. 2017KJ01).

Author information

Authors and Affiliations

  1. State Key Laboratory for Aquatic Animal Inspection and Testing, Lianyungang Customs, Lianyungang, 222042, Jiangsu Province, People’s Republic of China

    Ye Xu, Yi Zhou, Fengzhi Wang, Chao Ding, Jie Cao & Hongan Duan

Authors
  1. Ye Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yi Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fengzhi Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chao Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jie Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hongan Duan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hongan Duan.

Additional information

Editor: Tetsuji Okamoto

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xu, Y., Zhou, Y., Wang, F. et al. Development of two brain cell lines from goldfish and silver crucian carp and viral susceptibility to Cyprinid herpesivirus-2. In Vitro Cell.Dev.Biol.-Animal 55, 749–755 (2019). https://doi.org/10.1007/s11626-019-00402-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-019-00402-y

Keywords

Search

Navigation