Detection of the kinetoplastid Azumiobodo hoyamushi, the causative agent of soft tunic syndrome, in wild ascidians Halocynthia roretzi

Kumagai, Akira; Ito, Hiroshi C.; Sasaki, Ryo

doi:10.3354/dao02653

Cited by 12 publications

(6 citation statements)

References 12 publications

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“…Since AsSTS was first reported, efforts have been made to identify the causative agent, but recently, A. hoyamushi was found to be responsible [9,13]. In agreement with previous studies, the LAMP assay detected A. hoyamushi only in ascidians with AsSTS (Fig.…”

supporting

confidence: 87%

“…Grade 0 specimens were not available in Korea and thus Grade 1 specimens (non-diseased) were used as a control following the previous criteria. The collected ascidians were washed several times with 0.2 µm-filtered artificial seawater, tunics were dissected adjacent to the 2 siphons, chopped into small pieces (~0.2×0.2 cm) and placed in tubes containing 20 ml of sterilized artificial seawater, incubated at room temperature overnight, and directly examined under microscope the presence of flagellates as previously described [13]. To prepare ascidians genomic DNA for PCR and LAMP, the dissected ascidians prepared above were incubated with 10 ml of PBS at room temperature for 30 min.…”

mentioning

confidence: 99%

“…In the present study, to enhance the sensitivity of diagnostic methods, tunics adjacent to the siphons were chopped, incubated with PBS for only 30 min, heat-treated, and directly used as the template for the LAMP assay instead of using the extracted DNA by conventional methods from the overnight enrichment of the sliced tunics with seawater [13]. The LAMP assay conducted on heat-treated samples was found to be as efficient at detecting A. hoyamushi as DNA extracted using a commercial kit probably due to the ability of LAMP to better tolerate biological contaminants than PCR [14,15].…”

mentioning

confidence: 99%

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Development of Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Azumiobodo hoyamushi (Kinetoplastea)

et al. 2014

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Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.

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supporting

confidence: 87%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Development of Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Azumiobodo hoyamushi (Kinetoplastea)

et al. 2014

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“…An analysis of abundant OTUs revealed such associations for three neobodonid OTUs. The only parasitic neobodonid described so far is Azumiobodo (Hirose et al ., ; Kumagai et al ., ; Yazaki et al ., ), which was poorly represented within our samples (about 7 000 reads in total). Significantly more abundant were the novel parasite candidates: OTUs #4802, #3742, and #2083 (with 15 000, 29 000 and 45 000 reads, respectively).…”

Section: Resultsmentioning

confidence: 99%

“…2). Notably, Azumiobodo parasitizes on ascidians, a closely related group of benthic animals (Hirose et al, 2012;Kumagai et al, 2013), and another neobodonid, Cruzella marina, is a commensal in the ascidian intestine (Frolov and Malysheva, 2002).…”

Section: Ecological Interactionsmentioning

confidence: 99%

Neobodonids are dominant kinetoplastids in the global ocean

Flegontova

Flegontov

Malviya

et al. 2018

Environmental Microbiology

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Kinetoplastid flagellates comprise basal mostly free-living bodonids and derived obligatory parasitic trypanosomatids, which belong to the best-studied protists. Due to their omnipresence in aquatic environments and soil, the bodonids are of ecological significance. Here, we present the first global survey of marine kinetoplastids and compare it with the strikingly different patterns of abundance and diversity in their sister clade, the diplonemids. Based on analysis of 18S rDNA V9 ribotypes obtained from 124 sites sampled during the Tara Oceans expedition, our results show generally low to moderate abundance and diversity of planktonic kinetoplastids. Although we have identified all major kinetoplastid lineages, 98% of kinetoplastid reads are represented by neobodonids, namely specimens of the Neobodo and Rhynchomonas genera, which make up 59% and 18% of all reads, respectively. Most kinetoplastids have small cell size (0.8-5 µm) and tend to be more abundant in the mesopelagic as compared to the euphotic zone. Some of the most abundant operational taxonomic units have distinct geographical distributions, and three novel putatively parasitic neobodonids were identified, along with their potential hosts.

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Urochordates

Bakal¹,

Lewbart

2021

Invertebrate Medicine

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Detection of the kinetoplastid Azumiobodo hoyamushi, the causative agent of soft tunic syndrome, in wild ascidians Halocynthia roretzi

Cited by 12 publications

References 12 publications

Development of Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Azumiobodo hoyamushi (Kinetoplastea)

Development of Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Azumiobodo hoyamushi (Kinetoplastea)

Neobodonids are dominant kinetoplastids in the global ocean

Urochordates

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