A PCR-based diagnostic assay for the detection of Roseovarius crassostreae in Crassostrea virginica affected by juvenile oyster disease (JOD)

A.P. Maloy; B.J. Barber; K.J. Boettcher

Home/Archive/USGS

USGS · 70027582

A PCR-based diagnostic assay for the detection of Roseovarius crassostreae in Crassostrea virginica affected by juvenile oyster disease (JOD)

A.P. Maloy·B.J. Barber·K.J. Boettcher

View original source ↗Read source document ↗Export metadata ↓

Abstract

We have developed a PCR-assay for the diagnosis of juvenile oyster disease (JOD) based on the detection of Roseovarius crassostreae directly from affected oysters. Species-specific primers are used to amplify the 16S-23S rDNA internal transcribed spacer (ITS) of R. crassostreae, and confirmation of product identity is accomplished by restriction enzyme analysis. No false positives were obtained with either closely related bacterial species or from other DNAs present in oyster samples. The assay has the potential to detect as few as 10 cells of R. crassostreae per oyster when samples are taken from the inner valve surfaces of the animal. Inclusion of material from soft body surfaces is not necessary, and may reduce sensitivity approximately 10-fold. In a JOD-affected population, a positive PCR result was obtained from all oysters from which these bacteria were subsequently cultured. The assay also detected the presence of R. crassostreae in 2 oysters from which no R. crassostreae isolates were recovered. No R. crassostreae was detected by either PCR or bacteriology in oysters from a population that was not exhibiting JOD-signs. This assay is expected to advance regional disease management efforts and provide valuable insights into the disease process and epizootiology of JOD. ?? Inter-Research 2005.

Explore related subjects

Diseases of Aquatic Organisms Article

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

A.P. Maloy, B.J. Barber, K.J. Boettcher. 2005. A PCR-based diagnostic assay for the detection of Roseovarius crassostreae in Crassostrea virginica affected by juvenile oyster disease (JOD). https://pubs.usgs.gov/publication/70027582

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related USGS reports

USGS2026

First report of the flatworm Convolutriloba longifissura (Acoela) affecting a Caribbean coral species in a land-based nursery

Flatworms are widespread in the marine realm, but only a few species parasitize corals. Several of these are considered pests in the aquarium trade, and the invasive potential of these species is a concern. In the Caribbean there are no records to date of flatworms naturally occurring in coral nurseries and/or parasitizing corals in the wild. Here we report, for the first time, a flatworm identified as Convolutriloba longifissura affecting a Caribbean coral and include a phylogenetic and histological analyses. Flatworms were collected from nursery corals and artificial recruitment units (bioballs) suspended in the water column at Punta Cana reef. Sequencing and morphology of individuals from both indicate that C. longifissura is present in the wild as well as the nursery. Experiments were conducted to investigate the effect of flatworm exposure on signs of macroscopic stress (i.e. mucus production, paleness, bleaching, and tissue loss) on microfragments of Pseudodiploria clivosa . Standard histopathological techniques were applied to describe microscopic alterations of the coral exposed to C. longifissura. We found corals exposed to 30 individuals of C. longifissura had a 2.5-fold increased probability of developing macroscopic stress signs compared to controls after 14 d of exposure to different flatworm densities. While the actual impacts of these flatworms on coral restoration and microfragmentation programs or wild corals in the Caribbean remain to be determined, our results indicate that flatworms are no longer an exclusive problem of Indo-Pacific corals.

Punta Cana reef↗

USGS2025

Histological progression and bacterial load dynamics of Renibacterium salmoninarum in Chinook salmon Oncorhynchus tshawytscha

Renibacterium salmoninarum , the cause of bacterial kidney disease (BKD), severely impacts salmonid populations. Much of our understanding of the BKD pathology in salmonids comes from evaluating fatal infections in wild populations or spawning Pacific Northwest salmonids. Our study investigated the histological progression and bacterial load dynamics of R. salmoninarum infection in Chinook salmon following intraperitoneal injection to enhance understanding of the disease’s dynamics. Seventy presmolt salmon were injected with the ATCC-33209 isolate of R. salmoninarum and monitored over 10 wk. Histological signs of disease were observed in spleen and liver tissues at 1 wk post-injection and in kidney tissues at 4 wk post-injection, with signs of disease increasing over time. Additionally, histopathological analysis revealed splenic and hepatic capsulitis (coelomitis), individual hepatocyte necrosis, progressive granulomatous hepatitis, splenitis, and nephritis with necrosis. Gram staining confirmed the presence of Gram-positive bacteria within macrophages and extracellularly in infected tissues. Quantitative PCR revealed significant increases in bacterial loads in both kidney and liver tissues over the study period, with higher bacterial loads observed in the kidney tissue. Bacterial load was strongly correlated with disease stage, with peak bacterial burdens coinciding with the most severe histological changes. Our study provides a comprehensive account of R. salmoninarum infection and disease progression in Chinook salmon after intraperitoneal injection.

Diseases of Aquatic Organisms↗

USGS2024

Cytology in cnidaria using Exaiptasia as a model

A need exists for additional methods to examine cnidaria at the cellular level to aid our understanding of health, anatomy, and physiology of this important group of organisms. This need is particularly acute given that disease is emerging as a major factor in declines of ecologically important functional groups such as corals. Here we describe a simple method to process cnidarian cells for microscopic examination using the model organism Exaiptasia . We show that this organism has at least 18 cell types or structures that can be readily distinguished based on defined morphological features. Some of these cells can be related back to anatomic features of the animal both at the light microscope and ultrastructural level. The cnidome of Exaiptasia may be more complex than what is currently understood. Moreover, cnidarian cells, including some types of cnidocytes, phagocytize cells other than endosymbionts. Finally, our findings shed light on morphologic complexity of cell-associated microbial aggregates and their intimate intracellular associations. The tools described here could be useful for other cnidaria.

Diseases of Aquatic Organisms↗