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New host range for Hematodinium in southern Australia and novel tools for sensitive detection of parasitic dinoflagellates.

ABSTRACT: Hematodinium is a parasitic dinoflagellate and emerging pathogen of crustaceans. It preferably manifests in haemolymph of marine decapod crustaceans, killing a large variety of genera with significant impacts on fisheries worldwide. There is, however, evidence that some crustacean stocks harbor high prevalence, low intensity infections that may not result in widespread host mortality and are therefore hard to detect. The most widely used methods for detection of Hematodinium are conventional blood smears and polymerase chain reaction (PCR) against ribosomal RNAs. Blood smears demand a trained investigator, are labor intensive and not readily scalable for high-throughput sampling. PCRs only detect parasite DNA and can also suffer from false negatives and positives. In order to develop alternative detection tools for Hematodinium cells in decapod crustaceans we employed an immunological approach against a newly identified, abundant dinoflagellate-specific nuclear protein--Dinoflagellate/Viral NucleoProtein (DVNP). Both immunofluorescence assay (IFA) and Western blot methods against DVNP showed high sensitivity of detection. The Western blot detects Hematodinium parasites to levels of 25 parasites per milliliter of crustacean haemolymph, with the potential for sample pooling and screening of large samples. Using both PCR and these new tools, we have identified Hematodinium cells present in three new host crab taxa, at high prevalence but with no sign of pathogenesis. This extends the known range of Hematodinium to southern Australia.

SUBMITTER: Gornik SG

PROVIDER: S-EPMC3855790 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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New host range for Hematodinium in southern Australia and novel tools for sensitive detection of parasitic dinoflagellates.

Gornik Sebastian G SG Cranenburgh Andrea A Waller Ross F RF

PloS one 20131206 12

Hematodinium is a parasitic dinoflagellate and emerging pathogen of crustaceans. It preferably manifests in haemolymph of marine decapod crustaceans, killing a large variety of genera with significant impacts on fisheries worldwide. There is, however, evidence that some crustacean stocks harbor high prevalence, low intensity infections that may not result in widespread host mortality and are therefore hard to detect. The most widely used methods for detection of Hematodinium are conventional blo ...[more]

PMID: 24324829

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Project description:The sister phyla dinoflagellates and apicomplexans inherited a drastically reduced mitochondrial genome (mitochondrial DNA, mtDNA) containing only three protein-coding (cob, cox1, and cox3) genes and two ribosomal RNA (rRNA) genes. In apicomplexans, single copies of these genes are encoded on the smallest known mtDNA chromosome (6 kb). In dinoflagellates, however, the genome has undergone further substantial modifications, including massive genome amplification and recombination resulting in multiple copies of each gene and gene fragments linked in numerous combinations. Furthermore, protein-encoding genes have lost standard stop codons, trans-splicing of messenger RNAs (mRNAs) is required to generate complete cox3 transcripts, and extensive RNA editing recodes most genes. From taxa investigated to date, it is unclear when many of these unusual dinoflagellate mtDNA characters evolved. To address this question, we investigated the mitochondrial genome and transcriptome character states of the deep branching dinoflagellate Hematodinium sp. Genomic data show that like later-branching dinoflagellates Hematodinium sp. also contains an inflated, heavily recombined genome of multicopy genes and gene fragments. Although stop codons are also lacking for cox1 and cob, cox3 still encodes a conventional stop codon. Extensive editing of mRNAs also occurs in Hematodinium sp. The mtDNA of basal dinoflagellate Hematodinium sp. indicates that much of the mtDNA modification in dinoflagellates occurred early in this lineage, including genome amplification and recombination, and decreased use of standard stop codons. Trans-splicing, on the other hand, occurred after Hematodinium sp. diverged. Only RNA editing presents a nonlinear pattern of evolution in dinoflagellates as this process occurs in Hematodinium sp. but is absent in some later-branching taxa indicating that this process was either lost in some lineages or developed more than once during the evolution of the highly unusual dinoflagellate mtDNA.

Project description:Blastodinium is a genus of dinoflagellates that live as parasites in the gut of marine, planktonic copepods in the World's oceans and coastal waters. The taxonomy, phylogeny, and physiology of the genus have only been explored to a limited degree and, based on recent investigations, we hypothesize that the morphological and genetic diversity within this genus may be considerably larger than presently recognized. To address these issues, we obtained 18S rDNA and ITS gene sequences for Blastodinium specimens of different geographical origins, including representatives of the type species. This genetic information was in some cases complemented with new morphological, ultrastructural, physiological, and ecological data. Because most current knowledge about Blastodinium and its effects on copepod hosts stem from publications more than half a century old, we here summarize and discuss the existing knowledge in relation to the new data generated. Most Blastodinium species possess functional chloroplasts, but the parasitic stage, the trophocyte, has etioplasts and probably a limited photosynthetic activity. Sporocytes and swarmer cells have well-developed plastids and plausibly acquire part of their organic carbon needs through photosynthesis. A few species are nearly colorless with no functional chloroplasts. The photosynthetic species are almost exclusively found in warm, oligotrophic waters, indicating a life strategy that may benefit from copepods as microhabitats for acquiring nutrients in a nutrient-limited environment. As reported in the literature, monophyly of the genus is moderately supported, but the three main groups proposed by Chatton in 1920 are consistent with molecular data. However, we demonstrate an important genetic diversity within the genus and provide evidences for new groups and the presence of cryptic species. Finally, we discuss the current knowledge on the occurrence of Blastodinium spp. and their potential impact on natural copepod populations.

Dataset Information

New host range for Hematodinium in southern Australia and novel tools for sensitive detection of parasitic dinoflagellates.

Publications

New host range for Hematodinium in southern Australia and novel tools for sensitive detection of parasitic dinoflagellates.

Similar Datasets

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