Development of red sea bream iridovirus concentration method in seawater by iron flocculation

Kawato, Yasuhiko; Ito, Takafumi; Kamaishi, Takashi; Fujiwara, Atushi; Ototake, Mitsuru; Nakai, Toshiko; Nakajima, Kazuhiro

doi:10.1016/j.aquaculture.2015.08.016

Cited by 22 publications

(43 citation statements)

References 21 publications

(36 reference statements)

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“…The active viruses in the mucus might likely be able to be released into the cultured water and horizontally transmitted to other fish in the same population through direct contact or ingestion. If this is the case, assessment of viral loads in cultured water might be useful for SDDV detection and for early forecasts of the disease outbreaks (Kawato et al, 2016;Løvdal & Enger, 2002).…”

Section: Discussionmentioning

confidence: 99%

Detection of scale drop disease virus from non‐destructive samples and ectoparasites of Asian sea bass, Lates calcarifer

Charoenwai

Senapin

Dong

et al. 2020

Journal of Fish Diseases

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Scale drop disease virus (SDDV) is the causative agent of scale drop disease (SDD), a newly emerging disease of farmed Asian sea bass, Lates calcarifer in Singapore, Indonesia, Malaysia and Thailand (Gibson-Kueh et al., 2012; de Groof et al., 2015; Nurliyana et al., 2020; Senapin et al., 2019). SDDV is a double-stranded DNA virus, having an icosahedral shape (140-180 nm diameter), with a reported incomplete genome size of 124,244 bp (de Groof et al., 2015). The virus is currently classified as a novel Megalocytivirus, one of the five genera within the family Iridoviridae (de Groof et al., 2015). The major capsid protein encoding gene of SDDV had a ~64%-65% nucleotide identity to other members in the same genus including infectious spleen and kidney necrosis virus (ISKNV), red seabream iridovirus (RSIV) and turbot reddish body iridovirus (TBIV) and 73.28% identity to a newly identified

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Section: Discussionmentioning

confidence: 99%

Detection of scale drop disease virus from non‐destructive samples and ectoparasites of Asian sea bass, Lates calcarifer

Charoenwai

Senapin

Dong

et al. 2020

Journal of Fish Diseases

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“…Viral concentration using iron flocculation method was performed using the protocol previously described by Kawato et al (2016) with some modifications. Workflow of this method is illustrated in Fig.…”

Section: Methodsmentioning

confidence: 99%

“…With respect to waterborne transmission of fish pathogens, several studies employed various viral concentration methods from water for pathogen detection (For example, Haramoto et al (2007); Kawato et al (2016); Minamoto et al (2009); Nishi et al (2016)). The concept is one of the applications of environmental DNA (eDNA) which is nucleic acids extracted from environmental samples such as water, soil, and feces (Bass et al 2015; Gomes et al 2017).…”

Section: Introductionmentioning

confidence: 99%

“…The eDNA gives advantages in disease monitoring, control measure design, risk factor analysis and studies of viral survival nature (example review in Oidtmann et al (2018)). The work described by Kawato et al (2016) used an iron flocculation method to concentrate red sea bream iridovirus (RSIV) in a challenge model with Japanese amberjack ( Seriola quinqueradiata ). Results from that study showed that detection by qPCR of RSIV from fish-rearing water samples peaked more than five days before fish mortality occurred, suggesting potential benefit of using iron flocculation method for disease forecast.…”

Section: Introductionmentioning

confidence: 99%

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Concentration and quantification of Tilapia tilapinevirus from water using a simple iron flocculation coupled with probe-based RT-qPCR

Taengphu

Kayansamruaj

Kawato

et al. 2021

Preprint

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Tilapia tilapinevirus (also known as tilapia lake virus, TiLV) is an important virus responsible for die-off of farmed tilapia globally. Detection and quantification of the virus from environmental DNA/RNA (eDNA/eRNA) using pond water represents a potential, noninvasive routine approach for pathogen monitoring and early disease forecasting in aquaculture systems. Here, we report a simple iron flocculation method for viral concentration from water combined with a newly developed hydrolysis probe quantitative RT-qPCR method for detection and quantification of TiLV. The RT-qPCR method targeting a conserved region of TiLV genome segment 9 has a detection limit of 10 viral copies per uL of template. The method had a 100% analytical specificity and sensitivity for TiLV. The optimized iron flocculation method was able to recover 16.11 +/- 3.3% of virus from water samples spiked with viral cultures. During disease outbreak cases from an open-caged system and a closed hatchery system, both tilapia and water samples were collected for detection and quantification of TiLV. The results revealed that TiLV was detected from both clinically sick fish and asymptomatic fish. Most importantly, the virus was successfully detected from water samples collected from different locations in the affected farms e.g. river water samples from affected cages (8.50 x 102 to 2.79 x 104 copies/L) and fish-rearing water samples, sewage, and reservoir (4.29 x 102 to 3.53 x 103 copies/L) from affected and unaffected ponds of the hatchery. In summary, this study suggests that the eRNA detection system using iron flocculation coupled with probe based-RT-qPCR is feasible for concentration and quantification of TiLV from water. This approach might be useful for noninvasive monitoring of TiLV in tilapia aquaculture systems and facilitating appropriate decisions on biosecurity interventions needed.

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“…Several uncertain regions remained in the draft genome sequence, and hence, genome polishing was performed with a higher-purity viral genome prepared from virions shed from CEV-infected fish. Briefly, the rearing water of CEV-infected fish was filtered through a 0.4-μm polycarbonate filter (Advantec) and concentrated using the iron flocculation method ( 6 ) after dissolving 1% artificial seawater compound (Osaka Yakken). DNA was directly extracted from the flocculate-trapped filter using the DNeasy blood and tissue kit (Qiagen).…”

Section: Announcementmentioning

confidence: 99%

Complete Genome Sequence of Carp Edema Virus Isolated from Koi Carp

Mekata¹,

Kawato²,

Ito³

2021

Microbiol Resour Announc

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Development of red sea bream iridovirus concentration method in seawater by iron flocculation

Cited by 22 publications

References 21 publications

Detection of scale drop disease virus from non‐destructive samples and ectoparasites of Asian sea bass, Lates calcarifer

Detection of scale drop disease virus from non‐destructive samples and ectoparasites of Asian sea bass, Lates calcarifer

Concentration and quantification of Tilapia tilapinevirus from water using a simple iron flocculation coupled with probe-based RT-qPCR

Complete Genome Sequence of Carp Edema Virus Isolated from Koi Carp

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