Fine Structure of Clonally Propagated In Vitro Life Stages of a <i>Perkinsus</i> sp. Isolated from the Baltic Clam <i>Macoma balthica</i>

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

doi:10.1111/j.1550-7408.2001.tb00414.x

Cited by 47 publications

(64 citation statements)

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“…cultures have been established from a variety of tissues including hearts, visceral ganglia and gills (Kleinschuster & Swink 1993, La Peyre et al 1993, McLaughlin & Faisal 1998, Coss et al 2001a) but there is little information on the optimal source of parasites to use. The success rate in establishing continuous cultures can be used as a criterion for selecting a tissue as a source of parasites.…”

Section: Discussionmentioning

confidence: 99%

“…Results from this study contrast with reports of the transformation of zoospores into trophozoites and the subsequent growth in other Perkinsus spp. (Kleinschuster et al 1994, Coss et al 2001a. The role of zoospores in the life cycle of Perkinsus spp.…”

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confidence: 99%

“…The gill isolate G117 was further characterised as a new Perkinsus species, P. chesapeaki (Kotob et al 1999, McLaughlin et al 2000. 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.…”

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confidence: 99%

“…Cultures of the 3 Perkinsus spp. have been established from various sources including hearts, visceral ganglia and haemolymph of eastern oysters, gills and haemolymph of a softshell clam, and heart and haemolymph of baltic clams (Gauthier & Vasta 1993, Kleinschuster & Swink 1993, La Peyre et al 1993, McLaughlin & Faisal 1998, Coss et al 2001a. Parasite hypnospores isolated from infected eastern oyster tissues incubated in Ray's fluid thioglycollate medium (RFTM) were used to quickly establish P. marinus continuous cultures from different geographical regions , Bushek & Allen 1996b.…”

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confidence: 99%

“…P. marinus and P. chesapeaki were propagated in the culture medium JL-ODRP-1 originally formulated to resemble the composition of eastern oyster plasma (La Peyre et al 1993, McLaughlin & Faisal 1998. Commercial media including DMEM:Ham's F-12 and Leibowitz's L-15 medium, with various nutritional supplements, were used to culture P. marinus and P. andrewsi (Gauthier & Vasta 1993, Kleinschuster & Swink 1993, Kleinschuster et al 1994, Dungan & Hamilton 1995, Coss et al 2001a). Cultures of the 3 Perkinsus spp.…”

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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: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

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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Effects of plasma from bivalve mollusk species on the in vitro proliferation of the protistan parasite Perkinsus marinus

Gauthier

Vasta

2002

J. Exp. Zool.

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The in vitro culture of the Eastern oyster parasite Perkinsus marinus has provided a unique opportunity to examine its susceptibility to putative recognition and effector defense mechanisms operative in refractory bivalve species. In this study, we report the effect of supplementing the culture medium with plasma from: (1) uninfected to heavily infected Eastern oysters; (2) oyster species considered to be disease-resistant; and (3) bivalve mollusk species that are naturally exposed to the parasite but show no signs of disease. We also examined in vitro the interaction between hemocytes from Crassostrea virginica and C. gigas and P. marinus trophozoites. Our results revealed a significant decrease (32%) in proliferation of P. marinus in the presence of plasma from heavily infected C. virginica oysters. The inhibitory effects were less pronounced with plasma from moderately infected and uninfected oysters. In contrast, plasma from C. rivularis and C. gigas enhanced P. marinus proliferation. Proliferation was significantly reduced in media supplemented with plasma from Mytilus edulis, Mercenaria mercenaria, and Anadara ovalis. The highest inhibitory activity was apparent in M. edulis, for which 5% plasma-supplemented medium reduced growth by 35% relative to the controls. M. edulis active component(s) was heat-stable, yet pronase-sensitive. The significantly higher uptake of live P. marinus trophozoites by hemocytes from C. virginica, relative to those from C. gigas, suggests a certain level of specificity in the recognition/endocytosis of the parasite by its natural bivalve host species.

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Ciliary transition zone evolution and the root of the eukaryote tree: implications for opisthokont origin and classification of kingdoms Protozoa, Plantae, and Fungi

Cavalier-Smith

2021

Protoplasma

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I thoroughly discuss ciliary transition zone (TZ) evolution, highlighting many overlooked evolutionarily significant ultrastructural details. I establish fundamental principles of TZ ultrastructure and evolution throughout eukaryotes, inferring unrecognised ancestral TZ patterns for Fungi, opisthokonts, and Corticata (i.e., kingdoms Plantae and Chromista). Typical TZs have a dense transitional plate (TP), with a previously overlooked complex lattice as skeleton. I show most eukaryotes have centriole/TZ junction acorn-V filaments (whose ancestral function was arguably supporting central pair microtubule-nucleating sites; I discuss their role in centriole growth). Uniquely simple malawimonad TZs (without TP, simpler acorn) pinpoint the eukaryote tree's root between them and TP-bearers, highlighting novel superclades. I integrate TZ/ciliary evolution with the best multiprotein trees, naming newly recognised major eukaryote clades and revise megaclassification of basal kingdom Protozoa. Recent discovery of non-photosynthetic phagotrophic flagellates with genome-free plastids (Rhodelphis), the sister group to phylum Rhodophyta (red algae), illuminates plant and chromist early evolution. I show previously overlooked marked similarities in cell ultrastructure between Rhodelphis and Picomonas, formerly considered an early diverging chromist. In both a nonagonal tube lies between their TP and an annular septum surrounding their 9+2 ciliary axoneme. Mitochondrial dense condensations and mitochondrion-linked smooth endomembrane cytoplasmic partitioning cisternae further support grouping Picomonadea and Rhodelphea as new plant phylum Pararhoda. As Pararhoda/Rhodophyta form a robust clade on site-heterogeneous multiprotein trees, I group Pararhoda and Rhodophyta as new infrakingdom Rhodaria of Plantae within subkingdom Biliphyta, which also includes Glaucophyta with fundamentally similar TZ, uniquely in eukaryotes. I explain how biliphyte TZs generated viridiplant stellate-structures.

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Fine Structure of Clonally Propagated In Vitro Life Stages of a Perkinsus sp. Isolated from the Baltic Clam Macoma balthica

Cited by 47 publications

References 21 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

Effects of plasma from bivalve mollusk species on the in vitro proliferation of the protistan parasite Perkinsus marinus

Ciliary transition zone evolution and the root of the eukaryote tree: implications for opisthokont origin and classification of kingdoms Protozoa, Plantae, and Fungi

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