Histopathological characterization and in situ detection of Callinectes sapidus reovirus

Tang, Kathy F.J.; Messick, Gretchen A.; Pantoja, Carlos R.; Redman, Rita M.; Lightner, Donald V.

doi:10.1016/j.jip.2011.08.010

Cited by 20 publications

(6 citation statements)

References 6 publications

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“…The length of time to completion and the complexity of the assay (heated and unheated) promoted the development of a procedural checklist to reduce the potential for human error. The use of the unheated method contradicts the findings of most nonradioactive in situ hybridization methods previously reported by Mattsson et al (1993), Antonio et al (1998), or Tang et al (2011 by not requiring a target DNA heat-denaturation step. It is not clearly understood why the unheated method works.…”

Section: Discussioncontrasting

confidence: 66%

“…Oligonucleotide DNA probes for Perkinsus marinus (Moss et al 2006), P. chesapeaki (Reece et al 2008), Haplosporidium nelsoni (Stokes and Burreson 1995), Myxobolus cerebralis (Antonio et al 1998), Renibacterium salmoninarum (Mattsson et al 1993), and the Callinectes sapidus reovirus (Tang et al 2011) were purchased from Integrated DNA Technologies (Coralville, Iowa). Probe sequences for each of the pathogens are listed in Table 1.…”

Section: Methodsmentioning

confidence: 99%

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A Comparison of Two Methods for Colorimetric in situ Hybridization Using Paraffin‐Embedded Tissue Sections and Digoxigenin‐Labeled Hybridization Probes

Marcino

2013

J Aqua Anim Hlth

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Two methods for colorimetric in situ DNA probe hybridization (CISH) assays on paraffin-embedded tissue sections were compared. The heated method used heat (90-100°C) to denature DNA in the sample prior to probe hybridization, while the unheated method used a standard hybridization temperature of 42°C. Both procedures were tested on tissue samples that harbored the mollusk protozoan pathogens Perkinsus marinus, P. chesapeaki, or Haplosporidium nelsoni, the protozoan and bacterial fish pathogens Myxobolus cerebralis (myxosporidean) or Renibacterium salmoninarum (bacterial), or the crab viral pathogen Callinectes sapidus reovirus. Samples were fixed in either formalin or Davidson's fixative and embedded in paraffin for histological examination. The heated method is labor intensive and highly prone to human error, while the unheated method is less labor intensive and can be completed in a shorter period of time. Both methods yielded similar hybridization results. The use of complex and expensive prehybridization buffers did not improve the performances of the tested CISH assays. Prehybridization heat denaturation of DNA in assayed samples increased both assay duration and loss of samples but did not improve hybridization signals.

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Section: Discussioncontrasting

confidence: 66%

Section: Methodsmentioning

confidence: 99%

A Comparison of Two Methods for Colorimetric in situ Hybridization Using Paraffin‐Embedded Tissue Sections and Digoxigenin‐Labeled Hybridization Probes

Marcino

2013

J Aqua Anim Hlth

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“…However, one virus, CsRV1 (also called RLV for Reo‐Like Virus, or CsRV [Tang et al . ]), has been repeatedly found in association with mortality of captive crabs from Maryland and Virginia coastal bays (Johnson ; Messick & Kennedy ) and was recently discovered to be present in a majority of crabs dying in soft shell crab production systems in Maryland and the Gulf coast of Florida (Bowers et al . ).…”

Section: Introductionmentioning

confidence: 94%

“…), infects hemocytes, hemopoietic and connective tissues and causes pathology of tissues in association with hemocyte infiltration (Johnson ; Tang et al . ). The 55–60 nm icosahedral virions contain a double‐stranded RNA genome comprised of 12 segments.…”

Section: Introductionmentioning

confidence: 97%

“…Jahromi 1977;Johnson & Lightner 1988). However, one virus, CsRV1 (also called RLV for Reo-Like Virus, or CsRV [Tang et al 2011]), has been repeatedly found in association with mortality of captive crabs from Maryland and Virginia coastal bays (Johnson 1977;Messick & Kennedy 1990) and was recently discovered to be present in a majority of crabs dying in soft shell crab production systems in Maryland and the Gulf coast of Florida (Bowers et al 2010). The virus, which is fatal when injected into na€ ıve crabs (Bowers et al 2010), infects hemocytes, hemopoietic and connective tissues and causes pathology of tissues in association with hemocyte infiltration (Johnson 1977;Tang et al 2011).…”

Section: Introductionmentioning

confidence: 99%

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PCR‐based prevalence of a fatal reovirus of the blue crab, Callinectes sapidus (Rathbun) along the northern Atlantic coast of the USA

Flowers

Simmonds

Messick³

et al. 2015

Journal of Fish Diseases

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There is a need for more information on the relationship between diseases and fluctuations of wild populations of marine animals. In the case of Callinectes sapidus reovirus 1 (CsRV1, also known as RLV), there is a lack of baseline information on range, prevalence and outbreaks, from which to develop an understanding of population‐level impacts. An RT‐qPCR assay was developed that is capable of detecting 10 copies of the CsRV1 genome. In collaboration with state, federal and academic partners, blue crabs were collected from sites throughout the north‐eastern United States to assess the northern range of this pathogen. In addition, archived crab samples from the Chesapeake Bay were assessed for CsRV1 by RT‐qPCR and histology. PCR‐based assessments indicate that CsRV1 was present at all but one site. Prevalence of CsRV1 as assessed by RT‐qPCR was highly variable between locations, and CsRV1 prevalence varied between years at a given location. Mean CsRV1 prevalence as assessed by RT‐qPCR was >15% each year, and peak prevalence was 79%. The wide geographic range and highly variable prevalence of CsRV1 indicate that more study is needed to understand CsRV1 dynamics and the role the virus plays in blue crab natural mortality.

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Reovirus occurrence in mud crab farming systems and wild-caught brooders located in eastern coastal area of India

Ganesan

Baskaran

Raj³

et al. 2022

Aquacult Int

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Histopathological characterization and in situ detection of Callinectes sapidus reovirus

Cited by 20 publications

References 6 publications

A Comparison of Two Methods for Colorimetric in situ Hybridization Using Paraffin‐Embedded Tissue Sections and Digoxigenin‐Labeled Hybridization Probes

A Comparison of Two Methods for Colorimetric in situ Hybridization Using Paraffin‐Embedded Tissue Sections and Digoxigenin‐Labeled Hybridization Probes

PCR‐based prevalence of a fatal reovirus of the blue crab, Callinectes sapidus (Rathbun) along the northern Atlantic coast of the USA

Reovirus occurrence in mud crab farming systems and wild-caught brooders located in eastern coastal area of India

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