Description of <i>Perkinsus andrewsi</i> n. sp. Isolated from the Baltic Clam (<i>Macoma balthica</i>) by Characterization of the Ribosomal RNA Locus, and Development of a Species‐Specific PCR‐Based Diagnostic Assay

Coss, Cathleen A.; Robledo, José A. Fernández; Ruiz, Gregory M.; Vasta, Gerardo R.

doi:10.1111/j.1550-7408.2001.tb00415.x

Cited by 77 publications

(92 citation statements)

References 48 publications

(103 reference statements)

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“…In addition to the inability of P. qugwadi to enlarge in RFTM and to stain blue black with Lugol's solution, differences in morphology and considerable differences in DNA sequences of the ITS region of the ribosomal RNA gene complex between P. qugwadi and other Perkinsus spp. have been found (Coss et al 2001b, Casas et al 2002. In this study, the size of cultured cells incubated in RFTM increased significantly and the enlarged cells stained with Lugol's solution.…”

Section: Quantitation Of Growthmentioning

confidence: 78%

Section: Abstract: Perkinsus Atlanticus · Tapes Decussatus · In Vitrmentioning

confidence: 99%

“…Perkinsus parasites of infected Baltic clams Macoma balthica were cultured by Kleinschuster et al (1994) and Coss et al (2001a). The parasite cultured by Coss et al (2001b) was designated as a new Perkinsus species, P. andrewsi.Continuous cultures of Perkinsus chesapeaki and of P. andrewsi were established using procedures developed to propagate P. marinus. These procedures varied greatly among researchers in the type of medium used, the culture conditions and the method used for the establishment of P. marinus primary cultures (reviewed in La Peyre 1996).…”

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confidence: 99%

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Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus

Casas¹,

Peyre²,

Reece³

et al. 2002

Dis. Aquat. Org.

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Continuous in vitro cultures of the clam Tapes decussatus parasite Perkinsus atlanticuswere established from infected gill fragments, infected haemolymph and parasite hypnospores isolated from infected gill fragments following incubation in Ray's fluid thioglycollate medium (RFTM). No continuous cultures could be initiated from P. atlanticus zoospores. Cultures initiated from hypnospores yielded the highest percentage of continuous cultures (100%, 6/6), followed by cultures initiated from gill fragments (93%, 43/46) and from haemolymph (30%, 3/10). Failures to establish continuous cultures were due to microbial contamination. The source of parasite influenced the success rate, the time taken to establish cultures and the size of cultured cells. In vitro proliferation of parasite cells was mainly by vegetative multiplication. Zoosporulation, yielding motile biflagellated zoospores, was observed at a low frequency (<1% of dividing cells) in every culture. Morphology of cultured cells examined with light and transmission electron microscopy corresponded to that of P. atlanticus found in clam tissues. Cultured cells enlarged in RFTM and stained blue-black with Lugol's solution, which are characteristics of the Perkinsus species cells. DNA sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA gene complex matched those of P. atlanticus. All cultures were established in a medium designated JL-ODRP-2A that was similar in composition to the culture medium JL-ODRP-1 originally used to propagate Perkinsus marinus in vitro. Proliferation of P. atlanticus in vitro could be supported by the commercial culture medium (1:2 v/v) DME:Ham's F-12 with fetuin. KEY WORDS: Perkinsus atlanticus · Tapes decussatus · In vitro culture · Clam parasite · Ribosomal RNA gene complex · ITS · UltrastructureResale or republication not permitted without written consent of the publisher Dis Aquat Org 52: [217][218][219][220][221][222][223][224][225][226][227][228][229][230][231] 2002 spacer (ITS) region of the rRNA gene complex (Goggin 1994, de la Herrán et al. 2000, Robledo et al. 2000, Casas et al. 2002. Recently, Ordás & Figueras (1998) reported the in vitro culture of P. atlanticus from the haemolymph of an infected carpet shell clam. Identification of the cultured cells was based solely on their morphology (light and electron microscopy). Sequence analysis of the SSU small subunit rRNA gene, however, showed that those cultured cells did not correspond to a Perkinsus organism (Figueras et al. 2000). Continuous cultures of P. atlanticus, therefore, still need to be established.The development of continuous cultures of Perkinsus species can lead to a better understanding of this group of parasites . Cultures of the eastern oyster pathogen Perkinsus marinus, for example, have been used in a wide range of studies to address the parasite's environmental tolerance (Burreson et al. 1994, Dungan & Hamilton 1995, virulence , Bushek & Allen 1996a, Volety & Chu 1997, genetic composition (Reece et al. 1997, Reece et...

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Section: Quantitation Of Growthmentioning

confidence: 78%

Section: Abstract: Perkinsus Atlanticus · Tapes Decussatus · In Vitrmentioning

confidence: 99%

mentioning

confidence: 99%

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Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus

Casas¹,

Peyre²,

Reece³

et al. 2002

Dis. Aquat. Org.

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“…Two of the nine genes encoding rhomboid-like proteins in P. marinus have significant phylogenetic similarities with the apicomplexan rhomboids ROM1 and ROM6 and intriguingly early electron microscopy studies revealed the existence of rectilinear micronemes and the presence of a mitochondrion in Perkinsus spp. [70], suggesting that the P. marinus ROM1 and ROM6 localize to these organelles, as in T. gondii.…”

Section: Repertoire In Other Protozoansmentioning

confidence: 99%

New insights into parasite rhomboid proteases

Santos

Graindorge

Soldati‐Favre

2012

Molecular and Biochemical Parasitology

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a b s t r a c tThe rhomboid-like proteins constitute a large family of intramembrane serine proteases that are present in all branches of life. First studied in Drosophila, these enzymes catalyse the release of the active forms of proteins from the membrane and hence trigger signalling events. In protozoan parasites, a limited number of rhomboid-like proteases have been investigated and some of them are associated to pathogenesis. In Apicomplexans, rhomboid-like protease activity is involved in shedding adhesins from the surface of the zoites during motility and host cell entry. Recently, a Toxoplasma gondii rhomboid was also implicated in an intracellular signalling mechanism leading to parasite proliferation. In Entamoeba histolytica, the capacity to adhere to host cells and to phagocytose cells is potentiated by a rhomboid-like protease. Survey of a small number of protozoan parasite genomes has uncovered species-specific rhomboidlike protease genes, many of which are predicted to encode inactive enzymes. Functional investigation of the rhomboid-like proteases in other protozoan parasites will likely uncover novel and unexpected implications for this family of proteases.

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“…The species have been associated with mass mortalities of commercially important molluscs (Andrews & Hewatt 1957, da Ros & Canzonier 1985, Goggin & Lester 1987. Six species of this genus -P. marinus, P. olseni, P. atlanticus, P. qugwadi (insertae sedis), P. chesapeaki and P. andrewsi -have been described in molluscs from various parts of the world (Mackin et al 1950, Lester & Davis 1981, Azevedo 1989, Blackbourn et al 1998, McLaughlin et al 2000, Coss et al 2001. Recently, unidentified Perkinsus sp.…”

Section: Introductionmentioning

confidence: 99%

Effect of temperature and salinity on in vitro zoosporulation of Perkinsus sp. in Manila clams Ruditapes philippinarum

Ahn

Kim

2001

Dis. Aquat. Org.

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The effects of temperature and salinity on in vitro development of Perkinsus sp. prezoosporangia isolated from cultured Manila clams in Korea were investigated, and the difference in resistance to low temperature between prezoosporangia collected in winter and those collected in summer was compared. Temperature and salinity had significant effects on the development of prezoosporangia, and the developmental rates increased with increasing temperature and salinity. Prezoosporangia isolated in winter sporulated and released motile zoospores at 10°C, although the rates were significantly lower than those at 20 and 30°C. However, no prezoosporangia collected in summer sporulated at 10°C. Low salinities (≤10 ‰) had a significant negative effect on the development of prezoosporangia. A small number of prezoosporangia sampled in summer did sporulate at 5 ‰, but further developments including formation and release of zoospores were not observed. However, prezoosporangia sampled in winter and incubated at 5 ‰ released motile zoospores, although the rates were significantly lower than those at higher salinities. KEY WORDS: Perkinsus sp. · Ruditapes philippinarum · Manila clam · Prezoosporangia development · Temperature · Salinity Resale or republication not permitted without written consent of the publisherDis Aquat Org 48: [43][44][45][46] 2001 temperatures, salinities and chlorine concentrations. There are no data, however, concerning differences in tolerance of prezoosporangia to temperature and salinity as a function of the seasons at which the cells were sampled.In this study, therefore, the effects of temperature and salinity on in vitro development of Perkinsus sp. prezoosporangia isolated from cultured Manila clams in Korea were investigated, and the difference in tolerance to low temperatures between prezoosporangia collected in winter and in summer was compared. MATERIALS AND METHODSClams. Manila clams Ruditapes philippinarum were collected from cultured clam beds in an inner bay of the southern part of Korea in February and July 2000. The temperature and salinity in February were 9.8°C and 32.8 ‰, and in July were 25.0°C and 32.3 ‰, respectively. The experiments were conducted immediately after transferring the clams to the laboratory. The same experimental regimen was applied to both winter and summer samples.Preparation of prezoosporangia. Whole tissues of clams were incubated in fluid thioglycollate medium (FTM; Sigma) for 5 d in the dark at 30°C. After incubation, the tissues were minced and washed by centrifugation 4 times in sterile artificial seawater (ASW) adjusted to 30 ‰ and containing 200 units ml -1 penicillin/streptomycin (Sigma) and 500 units ml -1 nystatin (Sigma). The pellet was resuspended in ASW and filtered serially with sterilized gauze. Prezoosporangia were isolated from the filtrate with micropipettes under a stereomicroscope and inoculated into 24-well plates.Experimental regimen. The temperatures tested were 10, 20 and 30°C, and were controlled using an automatic temperature...

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Description of Perkinsus andrewsi n. sp. Isolated from the Baltic Clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species‐Specific PCR‐Based Diagnostic Assay

Cited by 77 publications

References 48 publications

Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus

Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus

New insights into parasite rhomboid proteases

Effect of temperature and salinity on in vitro zoosporulation of Perkinsus sp. in Manila clams Ruditapes philippinarum

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