Quantitative Polymerase Chain Reaction Assay for Largemouth Bass Virus

Getchell, Rodman G.; Groocock, Geoffrey H.; Schumacher, Vanessa; Grimmett, S G; Wooster, Gregory A.; Bowser, Paul R.

doi:10.1577/h07-009.1

Cited by 26 publications

(21 citation statements)

References 21 publications

(34 reference statements)

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“…A subsample (10) of the pooled tissue pieces of swim bladder, spleen, and posterior kidney collected in March 2007 were analyzed for the detection of largemouth bass virus (LMBV) by quantitative polymerase chain reaction analysis as described by Getchell et al (2007) at the Cornell University Aquatic Pathology Laboratory.…”

Section: Methodsmentioning

confidence: 99%

Mortality of Centrarchid Fishes in the Potomac Drainage: Survey Results and Overview of Potential Contributing Factors

et al. 2010

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Skin lesions and spring mortality events of smallmouth bass Micropterus dolomieu and selected other species were first noted in the South Branch of the Potomac River in 2002. Since that year morbidity and mortality have also been observed in the Shenandoah and Monocacy rivers. Despite much research, no single pathogen, parasite, or chemical cause for the lesions and mortality has been identified. Numerous parasites, most commonly trematode metacercariae and myxozoans; the bacterial pathogens Aeromonas hydrophila, Aeromonas salmonicida, and Flavobacterium columnare; and largemouth bass virus have all been observed. None have been consistently isolated or observed at all sites, however, nor has any consistent microscopic pathology of the lesions been observed. A variety of histological changes associated with exposure to environmental contaminants or stressors, including intersex (testicular oocytes), high numbers of macrophage aggregates, oxidative damage, gill lesions, and epidermal papillomas, were observed. The findings indicate that selected sensitive species may be stressed by multiple factors and constantly close to the threshold between a sustainable (healthy) and nonsustainable (unhealthy) condition. Fish health is often used as an indicator of aquatic ecosystem health, and these findings raise concerns about environmental degradation within the Potomac River drainage. Unfortunately, while much information has been gained from the studies conducted to date, due to the multiple state jurisdictions involved, competing interests, and other issues, there has been no coordinated approach to identifying and mitigating the stressors. This synthesis emphasizes the need for multiyear, interdisciplinary, integrative research to identify the underlying stressors and possible management actions to enhance ecosystem health.

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Section: Methodsmentioning

confidence: 99%

Mortality of Centrarchid Fishes in the Potomac Drainage: Survey Results and Overview of Potential Contributing Factors

et al. 2010

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“…and viruses (Bode et al 2004, Kocagoz et al 2005, Takahashi et al 2007, Tomaso et al 2007, Abril et al 2008, Kidd et al 2008. In recent years, fish disease diagnosticians have used this technique to identify and quantify bacterial, viral, and parasitic fish pathogens such as Aeromonas salmonicida, Flavobacterium columnare, Renibacterium salmoninarum, Henneguya ictaluri, largemouth bass virus, and recently Francisella piscicida in Norwegian cod (Balcázar et al 2007, Getchell et al 2007, Panangala et al 2007, Suzuki & Sakai 2007, Griffin et al 2008, Ottem et al 2008. The high sensitivity, high specificity, and short turnaround time for results make this technique an attractive replacement method for conventional diagnostic techniques (Espy et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Development of a real-time PCR assay for identification and quantification of the fish pathogen Francisella noatunensis subsp. orientalis

Soto¹,

Bowles²,

Fernandez³

et al. 2010

Dis. Aquat. Org.

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Members of the genus Francisella are small Gram-negative facultative intracellular bacteria that cause francisellosis in a wide variety of fish species worldwide. F. noatunensis subsp. orientalis has been recently described as a warm-water pathogen of tilapia Oreochromis spp. In this study, a quantitative real-time polymerase chain reaction (qPCR) TaqMan probe assay was developed to rapidly and accurately detect and quantify F. noatunensis subsp. orientalis from fish tissue. The target region of the assay was the F. tularensis iglC gene homologue previously found in F. noatunensis subsp. orientalis. Probe specificity was confirmed by the lack of signal and cross-reactivity with 12 common fish pathogens, 2 subspecies of F. tularensis, F. noatunensis subsp. noatunensis, and tilapia tissue. The range of linearity was determined to be 50 fg to 1.4 mg, and the lower limit of detection was 50 fg of DNA (equivalent to ~25 genome equivalents) per reaction. A similar sensitivity was observed with DNA extracted from a mixture of F. noatunensis subsp. orientalis and fish tissue. The assay was also able to detect and quantify F. noatunensis subsp. orientalis from the spleens of experimentally infected tilapia. No signal was observed in the control groups. In conclusion, we have developed a highly sensitive and specific assay that can be used for the specific identification and quantification of F. noatunensis subsp. orientalis. KEY WORDS: Francisella · Tilapia · qPCRResale or republication not permitted without written consent of the publisher

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“…In addition, adults of some herpetofauna have been shown to be carriers with no clinical signs of disease and with viral genomes sequestered in unsampled tissues or organs (Robert et al, 2007). Quantitative PCR is, however, a highly sensitive method (Kriger et al, 2006;Getchell et al, 2007) and we are confident that our results are an accurate reflection of the absence of FV3-like ranaviruses in L. clamitans larvae in our samples.…”

Section: Discussionmentioning

confidence: 68%

Surveying for Ranavirus in Green Frogs (Lithobates Clamitans) at Five Locations in Indiana

Winzeler¹,

Williams²,

Kimble³

2016

jnah

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Ranaviruses are an emerging pathogen within the United States that infects amphibians, reptiles, and fish. A Frog Virus 3-like (FV3) ranavirus has been detected at only two locations in Indiana; however, there have been few attempts to broadly sample for ranaviruses to determine their distribution across the state. This knowledge is necessary for the continued management and conservation of native amphibian populations. Our objective was to assess the occurrence of FV3-like ranaviruses in larval Green Frog (Lithobates clamitans) populations at five sites located in different regions of Indiana. Tissue samples were collected from 166 individuals and were assayed using both conventional and qPCR methods. We did not detect the presence of any FV3-like ranaviruses at any of the five sites with either PCR method, suggesting the possibility that at these sites, FV3-like ranaviruses may not be present. However, continued sampling should be carried out to monitor the status of the presence of ranaviruses in this portion of the Midwest.

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Quantitative Polymerase Chain Reaction Assay for Largemouth Bass Virus

Cited by 26 publications

References 21 publications

Mortality of Centrarchid Fishes in the Potomac Drainage: Survey Results and Overview of Potential Contributing Factors

Mortality of Centrarchid Fishes in the Potomac Drainage: Survey Results and Overview of Potential Contributing Factors

Development of a real-time PCR assay for identification and quantification of the fish pathogen Francisella noatunensis subsp. orientalis

Surveying for Ranavirus in Green Frogs (Lithobates Clamitans) at Five Locations in Indiana

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