Differentiation of BP-type baculovirus strains using in situ hybridization

Durand, Sébastien; Lightner, D. V.; Bonami, Jean-Robert

doi:10.3354/dao032237

Cited by 7 publications

(2 citation statements)

References 3 publications

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“…14,27,29 The hybridization signals confirm the presence of complementary DNA, which may be (but is not always) synonymous with the presence of a whole organism. ISH has been used for the differentiation of viral strains in crustaceans, 11 the detection of early stages of infection of myxosporidia in fish and their tubificid hosts, 3 the diagnosis of microsporidian-like Enterocytozoon bieneusi in immunosuppressed humans, 42 and the tracking of phage DNA through the intestinal wall of rodents. 34 The usefulness of the ISH assay for the detection of L. salmonae in the very early stages of infection in the intestine and heart was tested.…”

Section: Discussionmentioning

confidence: 99%

Localization of the Initial Developmental Stages of Loma salmonae in Rainbow Trout (Oncorhynchus mykiss)

et al. 2001

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Abstract. The intracellular microsporidian parasite Loma salmonae affects salmonids of the genus Oncorhynchus and is a significant cause of economic losses in pen-reared Chinook salmon (O. tshawytscha) in British Columbia. Loma salmonae infection is easily recognized by the xenomas that form in the gills, but early stages of infection are difficult to detect in histologic sections. In situ hybridization (ISH), using an L. salmonae-specific digoxigenin-labeled single-stranded DNA probe, was used to detect the parasite during the early stages of infection. Loma salmonae was detected in the gut mucosal epithelium as early as 24 hours postexposure (PE), and it localized in the lamina propria of the intestine within 24 hours of infection. After the parasite was detected in the lamina propria, dividing merogonic stages in infected cells in the heart were detected by ISH as early as 2 days PE, providing the first evidence of parasitaemia and hematogenous distribution of this parasite in infected blood cells. The parasites inside the infected cells appeared to be undergoing merogony as they passed through the heart, indicating that proliferation may start at the site of infection, before the parasite arrives to the gills for their final developmental phase. This is the first time that L. salmonae passage through the intestinal wall and migration to the heart has been visualized; however, the identity of the cells harboring the parasite has yet to be determined.

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Section: Discussionmentioning

confidence: 99%

Localization of the Initial Developmental Stages of Loma salmonae in Rainbow Trout (Oncorhynchus mykiss)

et al. 2001

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“…Indeed, a commercial DNA probe developed from HPV of Penaeus chinensis did not give positive hybridization results with HPV from Macrobrachium rosenbergii from Malaysia (Lightner et al 1994). To determine differences more precisely, molecular methods and ultimately DNA sequence comparisons are necessary, as has been done with Baculovirus penaei (Durand et al 1998) and for white spot syndrome virus (Lo et al 1999).…”

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confidence: 99%

Different reactions obtained using the same DNA detection reagents for Thai and Korean hepatopancreatic parvovirus of penaeid shrimp

Phromjai¹,

Sukhumsirichart²,

Pantoja³

et al. 2001

Dis. Aquat. Org.

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Hepatopancreatic parvovirus (HPV) can cause stunted growth and death in penaeid shrimp including Penaeus monodon. We used PCR primers and a commercial DNA probe designed from HPV of Penaeus chinensis (HPVchin) to examine HPV-infected Thai P. monodon (HPVmon). We found that the PCR primers produced a 732 bp DNA amplicon rather than the 350 bp amplicon obtained with HPVchin template and that the DNA probe gave weak to variable in situ DNA hybridization results. In addition, hybridization to PCR products from HPVmon was weak compared with hybridization with PCR products from HPVchin. By contrast, the 732 bp amplicon hybridized strongly with HPVmon-infected cells by in situ hybridization but not with uninfected shrimp tissue or other shrimp viruses, thus confirming its origin from HPVmon. Cloning, sequencing and analysis of the 732 bp amplicon showed that 696 bp (excluding the primer sequences) contained 47% GC content and had only 78% homology to 701 aligned bases from a 3350 bp DNA fragment of HPVchin from GenBank. These results explain why the reagents based on HPVchin gave a different PCR product and weak hybridization results with HPVmon, and they show that multiple primers or degenerate primers may be necessary for general detection of HPV varieties. Together with previously published information on the estimated total genome sizes for HPVchin (approximately 4 kb) and HPVmon (approximately 6 kb), these data support the contention that HPVchin and HPVmon are different varieties or species, in spite of their similar histopathology.

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Tetrahedral baculovirosis

Chong

2022

Aquaculture Pathophysiology

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Differentiation of BP-type baculovirus strains using in situ hybridization

Cited by 7 publications

References 3 publications

Localization of the Initial Developmental Stages of Loma salmonae in Rainbow Trout (Oncorhynchus mykiss)

Localization of the Initial Developmental Stages of Loma salmonae in Rainbow Trout (Oncorhynchus mykiss)

Different reactions obtained using the same DNA detection reagents for Thai and Korean hepatopancreatic parvovirus of penaeid shrimp

Tetrahedral baculovirosis

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