Mikrocytids Are a Broadly Distributed and Divergent Radiation of Parasites in Aquatic Invertebrates (original) (raw)

Molecular detection of the oyster parasite Mikrocytos mackini, and a preliminary phylogenetic analysis

2003

The protistan parasite Mikrocytos mackini, the causative agent of Denman Island disease in the oyster Crassostrea gigas in British Columbia, Canada, is of wide concern because it can infect other oyster species and because its life cycle, mode of transmission, and origins are unknown. PCR and fluorescent in situ hybridization (FISH) assays were developed for M. mackini, the PCR assay was validated against standard histopathological diagnosis, and a preliminary phylogenetic analysis of the M. mackini small-subunit ribosomal RNA gene (SSU rDNA) was undertaken. A PCR designed specifically not to amplify host DNA generated a 544 bp SSU rDNA fragment from M. mackini-infected oysters and enriched M. mackini cell isolates, but not from uninfected control oysters. This fragment was confirmed by FISH to be M. mackini SSU rDNA. A M. mackini-specific PCR was then designed which detected 3 to 4 × more M. mackini infections in 1056 wild oysters from Denman Island, British Columbia, than standard histopathology. Mikrocytos mackini prevalence estimates based on both PCR and histopathology increased (PCR from 4.4 to 7.4%, histopathology from 1.2 to 2.1%) when gross lesions were processed in addition to standard samples (i.e. transverse sections for histopathology, left outer palp DNA for PCR). The use of histopathology and tissue imprints plus PCR, and standard samples plus observed gross lesions, represented a 'total evidence' approach that provided the most realistic estimates of the true prevalence of M. mackini. Maximum parsimony and evolutionary distance phylogenetic analyses suggested that M. mackini may be a basal eukaryote, although it is not closely related to other known protistan taxa.

Aquatic Living Resources Microcell parasites of oysters : Recent insights and future trends

2004

Our understanding of the microcell oyster parasites of the genera Bonamia and Mikrocytos has expanded in recent years with the application of ultrastructural and especially molecular biological research approaches. Molecular phylogenetic analyses of SSU rRNA genes have united three species, Bonamia ostreae, Bonamia exitiosa, and Mikrocytos (now Bonamia) roughleyi, in a microcell clade within the Haplosporidia, supporting both early and recent ultrastructural observations. Ultrastructural and molecular phylogenetic evidence has emerged that Mikrocytos mackini, on the other hand, is a unique protist with unusual adaptations for a parasitic existence. DNA probes and polymerase chain reaction (PCR) assays promise new insights into the life cycles, transmission, and diversity of these organisms. The development of Ostrea edulis lines selected for B. ostreae resistance will increase the viability of aquaculture industries for this species and, combined with rapidly developing biotechnolog...

Phylogenetic, genomic, and biogeographic characterization of a novel and ubiquitous marine invertebrate-associated Rickettsiales parasite, Candidatus Aquarickettsia rohweri, gen. nov., sp. nov

The ISME Journal

Bacterial symbionts are integral to the health and homeostasis of invertebrate hosts. Notably, members of the Rickettsiales genus Wolbachia influence several aspects of the fitness and evolution of their terrestrial hosts, but few analogous partnerships have been found in marine systems. We report here the genome, phylogenetics, and biogeography of a ubiquitous and novel Rickettsiales species that primarily associates with marine organisms. We previously showed that this bacterium was found in scleractinian corals, responds to nutrient exposure, and is associated with reduced host growth and increased mortality. This bacterium, like other Rickettsiales, has a reduced genome indicative of a parasitic lifestyle. Phylogenetic analysis places this Rickettsiales within a new genus we define as "Candidatus Aquarickettsia." Using data from the Earth Microbiome Project and SRA databases, we also demonstrate that members of "Ca. Aquarickettsia" are found globally in dozens of invertebrate lineages. The coral-associated "Candidatus A. rohweri" is the first finished genome in this new clade. "Ca. A. rohweri" lacks genes to synthesize most sugars and amino acids but possesses several genes linked to pathogenicity including Tlc, an antiporter that exchanges host ATP for ADP, and a complete Type IV secretion system. Despite its inability to metabolize nitrogen, "Ca. A. rohweri" possesses the NtrY-NtrX two-component system involved in sensing and responding to extracellular nitrogen. Given these data, along with visualization of the parasite in host tissues, we hypothesize that "Ca. A. rohweri" reduces coral health by consuming host nutrients and energy, thus weakening and eventually killing host cells. Last, we hypothesize that nutrient enrichment, which is increasingly common on coral reefs, encourages unrestricted growth of "Ca. A. rohweri" in its host by providing abundant N-rich metabolites to be scavenged.

Microcell parasites of oysters: Recent insights and future trends

2004

Our understanding of the microcell oyster parasites of the genera Bonamia and Mikrocytos has expanded in recent years with the application of ultrastructural and especially molecular biological research approaches. Molecular phylogenetic analyses of SSU rRNA genes have united three species, Bonamia ostreae, Bonamia exitiosa, and Mikrocytos (now Bonamia) roughleyi, in a microcell clade within the Haplosporidia, supporting both early and recent ultrastructural observations. Ultrastructural and molecular phylogenetic evidence has emerged that Mikrocytos mackini, on the other hand, is a unique protist with unusual adaptations for a parasitic existence. DNA probes and polymerase chain reaction (PCR) assays promise new insights into the life cycles, transmission, and diversity of these organisms. The development of Ostrea edulis lines selected for B. ostreae resistance will increase the viability of aquaculture industries for this species and, combined with rapidly developing biotechnological approaches for studying host defenses and host-parasite interactions, will allow greater insight into the nature of phenomena such as resistance and tolerance to disease in oysters.

A rapid phenotype change in the pathogen Perkinsus marinus was associated with a historically significant marine disease emergence in the eastern oyster

Scientific Reports

The protozoan parasite Perkinsus marinus, which causes dermo disease in Crassostrea virginica, is one of the most ecologically important and economically destructive marine pathogens. The rapid and persistent intensification of dermo in the USA in the 1980s has long been enigmatic. Attributed originally to the effects of multi-year drought, climatic factors fail to fully explain the geographic extent of dermo’s intensification or the persistence of its intensified activity. Here we show that emergence of a unique, hypervirulent P. marinus phenotype was associated with the increase in prevalence and intensity of this disease and associated mortality. Retrospective histopathology of 8355 archival oysters from 1960 to 2018 spanning Chesapeake Bay, South Carolina, and New Jersey revealed that a new parasite phenotype emerged between 1983 and 1990, concurrent with major historical dermo disease outbreaks. Phenotypic changes included a shortening of the parasite’s life cycle and a tropism...

Host Range of the Mikrocytid Parasite Paramikrocytos canceri in Decapod Crustaceans

Pathogens

Mikrocytids are a widespread but rather neglected group of parasites of aquatic invertebrates. One such parasite is Paramikrocytos canceri—discovered to infect the antennal gland of the juvenile edible crab, Cancer pagurus, taken from several intertidal sites across the United Kingdom. To determine if this parasite is also present in other species of decapod crustaceans, we surveyed crabs (n = 330) across two contrasting sites in Pembrokeshire (UK). Using a histopathological approach, P. canceri infection was confirmed in variable numbers of edible crabs from both survey sites, 7–44%. No measurable signs of infection were encountered in four other co-located species, including European shore crabs (Carcinus maenas), Montagu’s crabs (Xantho hydrophilus), velvet swimming crabs (Necora puber) and broad-clawed porcelain crabs (Porcellana platycheles). These data imply that P. canceri has a more limited host range than suggested by molecular diagnosis alone.

Phylogeographic perspective on the distribution and dispersal of a marine pathogen, the oyster parasite Bonamia exitiosa

Marine Ecology Progress Series, 2015

The significance of infectious disease has intensified as our marine ecosystems are increasingly altered, with molluscan taxa being among the affected. One of the important pathogens to emerge in recent years, the oyster parasite Bonamia exitiosa, has a broad geographic distribution and has been found to infect a number of oyster species. In order to better understand how B. exitiosa achieved this wide distribution, a gene genealogy was constructed using internal transcribed spacer region ribosomal DNA sequencing data from across the host species range. The analysis revealed population structure in the form of 4 well-defined groups of sequences: 3 corresponding to geographic regions (temperate Atlantic and Pacific waters of the Southern Hemisphere, California, and the western Atlantic along the coast of the Americas) and the fourth geographically cosmopolitan. Inclusion of B. exitiosa sequences from New Zealand, Australia, and Argentina in the Southern Hemisphere group may reflect natural dispersal of the parasite via rafting with oyster hosts, whereas the California group may reflect limited anthropogenic movement of a host species, Ostrea lurida. The extensive geographic distribution of B. exitiosa parasites belonging to the cosmopolitan and Atlantic Coast groups may relate to both natural and anthropogenic dispersal of a single host, O. stentina, which is distributed from the eastern Americas to the Mediterranean and African coast to New Zealand-that is, in most regions where B. exitiosa has been found to occur.

Detection, Isolation, and Experimental Transmission ofMikrocytos mackini,a Microcell Parasite of Pacific OystersCrassostrea gigas(Thunberg)

Journal of Invertebrate Pathology, 1996

Denman Island disease, characterized by clinical signs of focal green lesions on the body surface or within the mantle, palps, and adductor muscle of Pacific oysters (Crassostrea gigas), is caused by a protozoan parasite of unknown taxonomic affiliations, Mikrocytos mackini. Detection of M. mackini was more sensitive and rapid by use of tissue imprints than histological sections. Of several isolation procedures investigated, centrifugation of homogenized infected tissues through a 15% sucrose solution enabled the isolation of the highest number of M. mackini with the lowest amount of oyster debris. Experimental transmission showed that oysters exposed to M. mackini by inoculation with isolated parasites had shorter prepatent periods and higher prevalences and intensities of infection than those incubated in homogenates from infected oysters (bath exposure) or those naturally exposed in the field. Experimental transmission was also used to propagate M. mackini in vivo in the laboratory year round. For the development of the disease, exposed oysters required prolonged incubation at low temperatures (about 10°C).

Molecular characterisation of a haplosporidian parasite infecting rock oysters Saccostrea cuccullata in north Western Australia

Journal of Invertebrate Pathology, 2007

A haplosporidian parasite was identiWed in rock oysters (Saccostrea cuccullata Born, 1778) from the Montebello Islands (latitude ¡20.4ЈS longitude 115.53ЈE) oV the northern coast of Western Australia by histopathological examination, PCR ampliWcation and DNA sequencing of a segment of the SSU region of the parasite's rRNA gene. An oligonucleotide probe was constructed from the parasite's SSU rRNA gene in order to conWrm its presence by in situ hybridisation. The parasite was disseminated throughout the gonad follicles of the host and to a lesser extent in the gills. The only parasite life stages thus far observed in this study were a uninucleate naked cell assumed to be a precursor to multinucleate plasmodial stages and a binucleate plasmodial stage. Whilst no parasite spores were detected in aVected rock oysters, a phylogenetic analysis of the SSU region of the parasite's rRNA gene indicates the parasite belongs to the genus Minchinia. A PCR and in situ hybridisation assay for the Minchinia sp. was used to identify haplosporidians described by Hine and Thorne [Hine, P.M.., Thorne, T., 2002. Haplosporidium sp. (Haplosporidia: Haplosporidiidae) associated with mortalities among rock oysters Saccostrea cuccullata in north Western Australia. Dis. Aquat. Organ. 51, 123-13], in archived rock oyster tissues from the same coastline. Crown

Richness and distribution of tropical oyster parasites in two oceans

Parasites can exert strong effects on population to ecosystem level processes, but data on parasites are limited for many global regions, especially tropical marine systems. Characterizing parasite diversity and distributions are the first steps towards understanding the potential impacts of parasites. The Panama Canal serves as an interesting location to examine tropical parasite diversity and distribution, as it is a conduit between two oceans and a hub for international trade. We examined metazoan and protistan parasites associated with ten oyster species collected from both Panamanian coasts, including the Panama Canal and Bocas del Toro. We found multiple metazoan taxa (pea crabs, Stylochus spp., Urastoma cyrinae). Our molecular screening for protistan parasites detected four species of Perkinsus (Perkinsus marinus, Perkinsus chesapeaki, Perkinsus olseni, Perkinsus beihaiensis) and several haplosporidians, including two genera (Minchinia, Haplosporidium). Species richness was higher for the protistan parasites than for the metazoans, with haplosporidian richness being higher than Perkinsus richness. Perkinsus species were the most frequently detected and most geographically widespread among parasite groups. Parasite richness and overlap differed between regions, locations and oyster hosts. These results have important implications for tropical parasite richness and the dispersal of parasites due to shipping associated with the Panama Canal.