Vaccination against cyprinid herpesvirus 2

Size: px

Start display at page:

Download "Vaccination against cyprinid herpesvirus 2"

Alban Sullivan
5 years ago
Views:

1 158, Bull. Eur. Ass. Fish Pathol., 33(5) 2013 Vaccination against cyprinid herpesvirus 2 Carassius auratus T. Ito 1 * and M. Ototake 2,3 2 Aquatic 3 Present address: Research Abstract Carassius auratus). However, in recent years, the virus has also been Carassius gibelio) and crucian carp (Carassius carassius was evaluated. The 50 l group, amplicons/polymerase chain reaction products appeared with decreased intensity indicat- Introduction Herpesviral haematopoietic necrosis (HVHN) (Carassius auratus) aquaculture in Japan since it 2008; Jung and Miyazaki, 1995). The disease is not only in Japan but also in the USA (Goodwin et al., 1999), Australia (Stephens et al., 2004), New Zealand (Hine et al., 2006) and the United - report (Jung and Miyazaki, 1995), and as this

2 Bull. Eur. Ass. Fish Pathol., 33(5) 2013, 159 as the species Cyprinid herpesvirus 2 (CyHV-2) Viruses. Although it was initially thought that this virus only demonstrated virulence towards Carassius gibelio) in China. Moreover CyHV-2 was also detected species in the Czech Republic. Furthermore, crucian carp (Carassius carassius) in Italy. These reports suggest that this virus may potentially Carassius. It is virus in cell culture has been limited (Jung and Miyazaki, 1995; Li and Fukuda, 2003). However, - tion and immersion using an in vitro propagated virus isolate (Ito et al., 2013). In this report, we Materials and methods Cell line minimum essential medium (MEM; Mediatech) (FBS; Equitech-Bio) and antibiotics (100 U penicillin ml -1 and 100 mg streptomycin ml -1 Virus and vaccine preparation The virus used in the present study was the CyHV-2 Saitama-1 (SaT-1) isolate which was times in GFF cells in 75cm 2 natant containing CyHV-2 were placed in 3.6 ml cryo tubes (Nunc and a TCID 50 - g supernatant, which contained TCID 50 ml -1 Vaccination procedure in the challenge trials. Edonishiki were bred - -

3 160, Bull. Eur. Ass. Fish Pathol., 33(5) were intraperitoneally inoculated with 0.1 ml anaesthetized by 2-phenoxyethanol diluted at the other two control groups received no injection. All experimental groups were kept in 60 l -1, Kyorin) once a day and were observed over a 21 day period to ensure that no detrimental cell cultures using InstraGene TM Matrix (Bio-Rad Puregene Tissue Kit (Qiagen). The PCR reaction mixture used was TaKaRa Ex Taq Hot Start - Virus re-isolation from e perimental sh 2 - Statistical analysis group and the respective control groups were determined by Fisher s exact test, P<0.01 values centage survival (RPS) was also determined by Challenge test An immersion challenge using the CyHV-2 Saitama-1 (SaT-1) isolate was conducted uti l -1 un-treated group) were immersed in diluted protocol. All 4 groups were kept in 60 l tanks as described previously and were observed over a 21 day period. The experimental design and Detection of viral D from sh by polymerase chain reaction (PCR) - Results jection accident. CyHV-2 DNA was not detected

4 Bull. Eur. Ass. Fish Pathol., 33(5) 2013, 161 Table 1. Group Challenge No. of samples Total length (cm) Body weight (g) Vaccinated Yes ± ± 4.34 Control Yes ± ± 2.93 Un-vaccinated Yes ± ± 5.89 Un-treated No ± ± 6.69 showed a normal appetite and similar activity are shown in Figure 1. The survival rates in and un-vaccinated groups P= Mortality within the respective negative control groups occurred between 3 and 11 days post-viral tality in the vaccinated group was delayed, occurring between 10 and 14 days post-viral challenge. A behavioral change (lethargy) was mortality among the vaccinated, control and non-vaccinated groups. There was no mortality in the non-challenged control group during the experimental period. Figure 1. ) Edonishiki were

5 162, Bull. Eur. Ass. Fish Pathol., 33(5) 2013 Figure 2. weight makers (Bioline, HyperLadder II); P, positive control; N, negative control. the viral challenge. Although viral DNA was - - died during the experiment (Figure 2). Viral observed in cultures inoculated with samples Discussion The present study clearly demonstrates the GFF cell cultures was recently published (Ito et al., 2013). From the vaccinated group only - inactivated vaccine was determined to be 57% group was delayed compared to the control and - (Jung and Miyazaki, 1995). While in our previ-

6 Bull. Eur. Ass. Fish Pathol., 33(5) 2013, 163 Table 2. Group Survival rate postchallenge (%) (Surviving /Challenged) PCR Dead sh Virus reisolation Surviving sh PCR Virus reisolation Vaccinated 57 (8/14) 6/6 5/6 8/8 0/8 Control 0 (0/15) 15/15 15/ Un-vaccinated 0 (0/15) 15/15 14/ Un-treated 100 (15/15) - - 0/15 NT Acknowledgement clinical signs due to CyHV-2 experimental in- tion experiment a behavioral change (lethargy) (Ito et al., 2013). These indistinct clinical signs sub-clinical carriers resulting in transport not cell culture are low compared with other CyHVs such as CyHV-1 or CyHV-3 although the sus- in vitro must be investigated. Furthermore, trialing the vaccine on a larger scale to establish the search Agency in Japan. We thank Y. Maeno References Available at html (in Japanese) (Accessed June 24, 2013). Chang PH, Lee SH, Chiang HC and Jong Carassius auratus in Taiwan. Fish Pathology 34, M, Pokorová D and Knytl M (2012). Massive Carassius gibelio in the upper Elbe basin associated with herpesviral hematopoietic necrosis (CyHV- 2). Diseases of Aquatic Organisms 102, Fichi G, Cardeti G, Cocumelli C, Vendramin N, herpesvirus 2 in association with an Aeromonas sobria Carassius

7 164, Bull. Eur. Ass. Fish Pathol., 33(5) 2013 carassius (L.), in Italy. Journal of Fish Diseases 36, Goodwin AE, Khoo L, LaPatra SE, Bonar C, Key DW, Garner M, Lee MV and Hanson herpesvirus (Cyprinid Herpesvirus 2) in Journal of Aquatic Animal Health 18, JG (1998). A viral epizootic in cultured putative herpesvirus etiology. Journal of 10, Hine PM, Tham K-M and Morrison R (2006). Cyprinid herpesvirus 2 in New Zealand Surveillance 33, 3-5. Ito T, Kurita J, Ozaki A, Sano M, Fukuda H herpesvirus 2 (CyHV-2) in cell culture and Carassius auratus. Diseases of Aquatic Organisms 105, Carassius auratus (L.), in the UK. Journal of Fish Diseases 30, Jung SJ and Miyazaki T (1995). Herpesviral Carassius auratus (L.). Journal of Fish Diseases 18, Li X and Fukuda H (2003). culture herpesvirus. Journal of Shanghai Fisheries University 12, Luo YZ, Lin L, Liu Y, Wu ZW, Gu ZM, Li LJ and Yuan JF (in press) Haematopoietic Carassius gibelio (Bloch), associated with Cyprinid herpesvirus 2. Journal of Fish Diseases doi Reed LJ and Muench H (1938). A simple method American Journal of Epidemiology 27, Stephens FJ, Raidal SR and Jones B (2004). (Carassius auratus) associated with an agent morphologically similar to herpesvirus. Australian Veterinary Journal 82, polymerase chain reaction assay to detect Journal of Aquatic Animal Health 21, Wang L, He J, Liang L, Zheng X, Jia P, Shi X, Lan W, Xie J, Liu H and Xu P (2012). Mass mortality caused by Cyprinid Herpesvirus 2 (CyHV-2) in Prussian carp (Carassius gibelio) in China. Bulletin of the European Association of Fish Pathologists 32, Wu T, Ding Z, Ren M, An L, Xiao Z, Liu P, Gu W, Meng Q and Wang W (2013). The histo- Carassius gibelio) in mainland China associated with cyprinid herpesvirus 2 (CyHV-2). Aquaculture , 8-13.

Virulence of viral haemorrhagic septicaemia virus (VHSV) genotype III in rainbow trout

DOI 10.1186/s13567-015-0303-z Veterinary Research RESEARCH ARTICLE Open Access Virulence of viral haemorrhagic septicaemia virus (VHSV) genotype III in rainbow trout Takafumi Ito 1*, Jun Kurita 1, Koh