Seasonality in the infection and invasion of Marteilioides chungmuensis in the Pacific oyster Crassostrea gigas

Tun, Kay Lwin; Shimizu, Yasuko; Yamanoi, Hideo; Yoshinaga, Tomoyoshi; Ogawa, Kazuo

doi:10.3354/dao01924

Cited by 9 publications

(3 citation statements)

References 15 publications

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“…However, sensitivity of this PCR diagnosis is not sufficient for practical use and nested-PCR using another primer pair, OPF3/OPR3, is recommended to increase sensitivity (Itoh et al, 2003b). This PCR protocol was used for detection of the parasite in male oysters (Itoh et al 2004a), diagnosis of various bivalves (Itoh et al 2004b;Limpanot et al 2013;Sühnel et al 2014) and determination of invasion period of the parasite (Tun et al 2008b). In order to morphologically characterise M. chungmuensis cells outside oyster oocytes, in situ hybridisation probes for M. chungmuensis, MCSP-01, MCSP-05 and 6-R, were developed (Itoh et al, 2003a).…”

Section: 12molecular Detection Of Marteilia Sydneyimentioning

confidence: 99%

Marteilia spp. parasites in bivalves: A revision of recent studies

Carrasco

Green

Itoh

2015

Journal of Invertebrate Pathology

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Section: 12molecular Detection Of Marteilia Sydneyimentioning

confidence: 99%

Marteilia spp. parasites in bivalves: A revision of recent studies

Carrasco

Green

Itoh

2015

Journal of Invertebrate Pathology

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“…우리나라 바지락 생산량은 1990년 차지한다 . 국내의 바지락 생산량 감소 원 인은 생리적요인 (Keino et al 2005;Yan et al, 2006), 질병 요인 (Park and Choi, 2001;Allam et al, 2002;Ngo et al, 2003;Park et al, 2006;Tun et al, 2008;Park et al, 2010) 및 어장 관리적 요인 (Lee et al, 1996) …”

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Conditioning of Manila clam Ruditapes philippinarum (Adams & Reeve, 1850) using recirculation system: I. Induction of the gametogenesis using water temperature elevation

Lee¹,

Park²,

Choi³

2014

The Korean Journal of Malacology

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Gonad maturation of Manila clam, Ruditapes philippinarum was induced in this study using a recirculation system over 8 weeks in early spring. Clams used in the experiment were collected in 15 th April 2010 from the west coast of Korea, as the surface water temperature remained 11°C. To induce gametogenesis and subsequent maturation seawater temperature was elevated 1°C per day over 10 days to reach 20°C. For the experiment, clams were raised in 120 L quadrangle tank maintained with re-circulated seawater system over 57 days. Water quality parameters including the water temperature, salinity dissolved oxygen, ammonium ion and nitrate levels in the tanks were monitored daily. Mixture of concentrated microalgae including Tetraselmis, Isochrysis, Pavlova and Thalassiosira weissflogii was supplied to clams twice a day, and quantity of the daily ration was adjusted as 3% of clam body dry weight. Histology was applied to examine gonad maturation. Daily monitoring of the water quality parameters indicated that the recirculation system supplied suitable environment to Manila clam; the nitrogenous components stayed below toxic levels (< 0.2 mg/L). At the beginning of the study, clams were mostly in early developing stage. As the seawater temperature reached 20°C, 10 days after the experiment, 20% of clams reached late development at 12 days. First ripe clams were observed at 42 days and 40% of clams were in ripe and ready for spawning at the end of study, 57 days after the experiment. In this study, gametogenesis of Manila clam was successfully induced by elevating water temperature and supplying commercially produced microalgae in a recirculation tank system.

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“…The use of polymerase chain reaction (PCR) assays for detecting parasites of commercially important bivalves has progressed from initial development (Anderson et al 1995, Stokes et al 1995, Robledo et al 1998, Penna et al 2001, Carnegie et al 2003 to implementation in field studies (Lyons et al 2005, Carnegie et al 2008, Tun et al 2008, Ford et al 2009a and is beginning to be used for seed certification (e.g. Ford et al 2009b).…”

Section: Introductionmentioning

confidence: 99%

Quantitative PCR assay to determine prevalence and intensity of MSX (Haplosporidium nelsoni) in North Carolina and Rhode Island oysters Crassostrea virginica

Wilbur¹,

Ford²,

Gauthier³

et al. 2012

Dis. Aquat. Org.

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The continuing challenges to the management of both wild and cultured eastern oyster Crassostrea virginica populations resulting from protozoan parasites has stimulated interest in the development of molecular assays for their detection and quantification. For Haplosporidium nelsoni, the causative agent of multinucleated sphere unknown (MSX) disease, diagnostic evaluations depend extensively on traditional but laborious histological approaches and more recently on rapid and sensitive (but not quantitative) end-point polymerase chain reaction (PCR) assays. Here, we describe the development and application of a quantitative PCR (qPCR) assay for H. nelsoni using an Applied Biosystems TaqMan ® assay designed with minor groove binder (MGB) probes. The assay was highly sensitive, detecting as few as 20 copies of cloned target DNA. Histologically evaluated parasite density was significantly correlated with the quantification cycle (C q ), regardless of whether quantification was categorical (r 2 = 0.696, p < 0.0001) or quantitative (r 2 = 0.797, p < 0.0001). Application in field studies conducted in North Carolina, USA (7 locations), revealed widespread occurrence of the parasite with moderate to high intensities noted in some locations. In Rhode Island, USA, application of the assay on oysters from 2 locations resulted in no positives. KEY WORDS: MSX · Oysters · qPCR · Diagnostic assay · Histology · Parasite density Resale or republication not permitted without written consent of the publisherDis Aquat Org 102: [107][108][109][110][111][112][113][114][115][116][117][118] 2012 history stages in atypical hosts, vectors, and/or environmental samples, and they provide a sensitive tool for detecting the pathogens at early stages of infection.Traditional PCR assays analyze end-point amplification products that generally require post-amplification handling for quantification, reducing their utility for some questions. These limitations have prompted interest in real-time or quantitative PCR (qPCR) assays, which have followed a trajectory similar to end-point PCR, from initial development as a diagnostic tool (Day et al. 2000, Yarnall et al. 2000, Audemard et al. 2004, Gauthier et al. 2006, Lyons et al. 2006, De Faveri et al. 2009) to application for understanding the ecology of the parasite in the environment, and mechanisms of host−pathogen interactions (Audemard et al. 2006, Marty et al. 2006, Gast et al. 2008, Liu et al. 2009). These applications have been made possible by the fact that qPCR assays permit the absolute or relative quantification of the target. For species of parasites that can be propagated in vitro, a qPCR assay can be calibrated using counts of cultured parasites (Yarnall et al. 2000, Audemard et al. 2004, Gauthier et al. 2006, De Faveri et al. 2009) to convert relative to absolute abundance. The inability to propagate some important bivalve pathogens complicates the development of qPCR assays for those pathogens. In such cases, calibration can be achieved by using semi-quantitative rating...

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Seasonality in the infection and invasion of Marteilioides chungmuensis in the Pacific oyster Crassostrea gigas

Cited by 9 publications

References 15 publications

Marteilia spp. parasites in bivalves: A revision of recent studies

Marteilia spp. parasites in bivalves: A revision of recent studies

Conditioning of Manila clam Ruditapes philippinarum (Adams & Reeve, 1850) using recirculation system: I. Induction of the gametogenesis using water temperature elevation

Quantitative PCR assay to determine prevalence and intensity of MSX (Haplosporidium nelsoni) in North Carolina and Rhode Island oysters Crassostrea virginica

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