Concentrations of a Koi herpesvirus (KHV) in tissues of experimentally-infected Cyprinus carpio koi as assessed by real-time TaqMan PCR

Gilad, Oren; Yun, Susan; Zagmutt-Vergara, Francisco J.; Leutenegger, Christian M.; Bercovier, Hervé; Hedrick, Ronald P.

doi:10.3354/dao060179

Cited by 303 publications

(442 citation statements)

References 29 publications

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“…Based on the fact that in the presently reported study KHV was detected in the specimens collected from Russian and Atlantic sturgeon it seems that restocking should be accompanied by a control management, focusing especially on viruses. This is crucial, as by use of diagnostic methods, such as nested PCR or realtime PCR, even 10 fg of viral DNA, corresponding to 5-10 KHV genomic equivalents measured by realtime PCR (Gilad et al 2004), can be detected. The use of molecular tools for KHV detection can provide an efficient control of invasive species introduced into local waters.…”

Section: Discussionmentioning

confidence: 99%

Koi Herpes Virus: Do Acipenserid Restitution Programs Pose a Threat to Carp Farms in the Disease-Free Zones?

Kempter¹,

Sadowski²,

Schütze³

et al. 2009

Acta Icth et Piscat

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Background. Sturgeons have long been extinct in Polish inland waters. A substantial effort has recently been put into their restitution, covering the drainage areas of two major Polish rivers, the Oder and the Vistula. The stocked fishes are clinically healthy, but very little is known about their potential to transmit viral diseases including koi herpes virus (KHV) to healthy fishes of other species, which may pose a threat to the disease-free zones. This study was intended to determine if sturgeons could be asymptomatic carriers of KHV. Materials and Methods. A total of 29 sturgeons (two species; length 8-37 cm) originating from fish farms in northern Poland with a known KHV history in common carp or koi in the area were examined: 15 Russian sturgeons, Acipenser gueldenstaedtii, with clinical signs of a disease and 14 asymptomatic Atlantic sturgeons, A. oxyrinchus. The former were sent to the laboratory alive while the latter were sent fixed in ethanol. As it is required for detection of a latent KHV infection in acipenserids, two independent procedures were applied. The preliminary results were obtained using PCR. Those findings were subsequently confirmation by nested PCR. The latter procedure consists of sequence analysis of PCR products and direct detection of KHV infected cells in tissue materials by in-situ hybridization on nucleic acid level or indirect immunofluorescence on KHV protein level. Results. KHV genome parts were found in nine Russian sturgeons and four Atlantic sturgeons. Comparison of PCR results obtained from three primer pairs used for KHV diagnostic in sturgeon showed that those designed by Bercovier et al. were most sensitive and robust for this purpose. In order to confirm the presence of viral particles the most useful method was in-situ hybridization (ISH), allowing the detection of KHV in gill samples obtained from live sturgeons.Conclusion. This preliminary study shows that sturgeons can be carriers of KHV. Therefore a viral diagnostics is highly recommended not only for sturgeons obtained from the environment but also for fertilized eggs, fry, and fish intended for re-stocking measurements of inland waters.

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Section: Discussionmentioning

confidence: 99%

Koi Herpes Virus: Do Acipenserid Restitution Programs Pose a Threat to Carp Farms in the Disease-Free Zones?

Kempter¹,

Sadowski²,

Schütze³

et al. 2009

Acta Icth et Piscat

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“…Extracted DNA was then purified using a QIAquick DNA Purification Kit (Qiagen), which yielded 50 ml of purified DNA. CyHV-3 DNA and a known single-copy gene in common carp (C. carpio glucokinase gene) as an internal control were quantified using TaqMan real-time PCR, according to Gilad et al (2004). For each TaqMan assay of CyHV-3 and the glucokinase gene, reaction mixtures of 10 ml of TaqMan Gene Expression Master Mix (Applied Biosystems, Foster City, CA, USA), 900 nM of each primer pair, 125 nM of a TaqMan probe and 2 ml of sample DNA in a 20-ml volume were run in triplicate for all samples using the StepOnePlus real-time PCR system (Applied Biosystems).…”

Section: Measurement Of Serum Testosteronementioning

confidence: 99%

Transmission dynamics of an emerging infectious disease in wildlife through host reproductive cycles

et al. 2010

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Emerging infectious diseases are major threats to wildlife populations. To enhance our understanding of the dynamics of these diseases, we investigated how host reproductive behavior and seasonal temperature variation drive transmission of infections among wild hosts, using the model system of cyprinid herpesvirus 3 (CyHV-3) disease in common carp. Our main findings were as follows: (1) a seroprevalence survey showed that CyHV-3 infection occurred mostly in adult hosts, (2) a quantitative assay for CyHV-3 in a host population demonstrated that CyHV-3 was most abundant in the spring when host reproduction occurred and water temperature increased simultaneously and (3) an analysis of the dynamics of CyHV-3 in water revealed that CyHV-3 concentration increased markedly in breeding habitats during host group mating. These results indicate that breeding habitats can become hot spots for transmission of infectious diseases if hosts aggregate for mating and the activation of pathogens occurs during the host breeding season.

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“…Subsequently, DNA can be detected in the gill, intestinal tract, spleen and liver, but not in the brain [29]. Virus is most abundant in gill, kidney, and spleen during the course of infection [27]. KHV DNA has been detected in the environmental water before, during, and after an outbreak of the disease [33,52].…”

Section: Transmissionmentioning

confidence: 99%

“…Several diagnostic tests based on PCR [6,25,29,89], nested PCR [7,20], real-time PCR [27] and loop-mediated isothermal amplification (LAMP) [31,75,87,88] have been developed for detection of KHV. A highly sensitive PCR method for detection of TK gene of KHV is able to detect 10 fg of KHV DNA [6].…”

Section: Molecular Detection Of Viral Dnamentioning

confidence: 99%

“…Sensitivity of some of these diagnostic assays has been compared [8]. The real-time PCR recognizing KHV DNA fragment [27] with an internal [36] and external control system is considered as the ''gold standard'' for detection and absolute virus quantification. Nested PCR [7] and Bercovier's PCR [6] also have the same sensitivity as the ''gold standard'' which correspond to 10 fg DNA or 1-5 genomic KHV equivalents.…”

Section: Molecular Detection Of Viral Dnamentioning

confidence: 99%

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Koi Herpes Virus: A Review and Risk Assessment of Indian Aquaculture

et al. 2012

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Common carp (Cyprinus carpio) is a widely cultivated freshwater fish for human consumption, while koi carp, is a farmed colored sub species of common carp used for ornamental purposes. Since 1998, both common carp and koi carp are severely affected by a viral disease called as Koi herpes virus disease (KHVD). This disease is caused by Koi herpes virus (KHV), also known as cyprinid herpes virus-3. The virus causes interstitial nephritis and gill necrosis in carps, so it is also termed as carp interstitial nephritis and gill necrosis virus. KHV is a double stranded icosahedral DNA virus belonging to family Alloherpesviridae, with a genome size of 295 kbp, larger than any member of Herpesviridae. The viral genome encodes 156 potential protein coding open reading frames. Each virion consists of forty structural proteins, which are classified as capsid (3), envelope (13), tegument (2) and unclassified (22) structural proteins. Diagnosis of KHVD is mainly based on detection of viral DNA by polymerase chain reaction amplification using specific primers or loop mediated isothermal amplification. Temperature dependent latent infection is unique to KHV; and carrier fish are often not detected, thereby possibly resulting in spread of this pathogen to newer areas. The disease is now known to occur in, or has been recorded from at least 26 different countries of the world. Fortunately, KHVD has not been reported from India or from Indian major carps. To monitor the disease status of the country, a total of 254 fish samples collected from different parts of India were screened by PCR for the presence of KHV. None of the tested samples were found to be positive for KHV. These results demonstrate that tested samples from different parts of India were apparently free from KHV. Preliminary risk assessment of KHV suggest that in the event of unrestricted importation of koi carps into our country, there is a higher probability of risk to aquaculture as compared to natural waters. So there is strong need to develop diagnostic capabilities and launch surveillance programmes for KHV in India.

show abstract

Concentrations of a Koi herpesvirus (KHV) in tissues of experimentally-infected Cyprinus carpio koi as assessed by real-time TaqMan PCR

Cited by 303 publications

References 29 publications

Koi Herpes Virus: Do Acipenserid Restitution Programs Pose a Threat to Carp Farms in the Disease-Free Zones?

Koi Herpes Virus: Do Acipenserid Restitution Programs Pose a Threat to Carp Farms in the Disease-Free Zones?

Transmission dynamics of an emerging infectious disease in wildlife through host reproductive cycles

Koi Herpes Virus: A Review and Risk Assessment of Indian Aquaculture

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