The role of alternate hosts in the ecology and life history of Hematodinium sp., a parasitic dinoflagellate of the blue crab (Callinectes sapidus) (original) (raw)
Related papers
Molecular Detection of Hematodinium Sp. Infecting the Blue Crab, Callinectes Sapidus
Journal of Shellfish Research, 2007
Species of Hematodinium are endoparasitic dinoflagellates of crustaceans. Certain stages of the parasites can be very difficult to detect in the hemolymph of their hosts, because the trophic stages resemble hemocytes, and they can occur at relatively low densities, making diagnosis by microscopy difficult. We developed a polymerase chain reaction (PCR) assay to detect the Hematodinium sp. infecting the blue crab, Callinectes sapidus, based on the amplification of the parasite's first internal transcribed spacer region (ITS1) of the ribosomal RNA (rRNA) gene complex. The PCR assay was combined with a restriction endonucleases digestion (Bsg I) of the amplification products to differentiate between different forms of Hematodinium from different hosts. The assay had a limit of detection equivalent to 0.3 parasites per 100-mL hemolymph. In addition, two oligonucleotide DNA probes were designed to target the 18S rRNA gene sequence of the parasite, facilitating detection in situ in crustacean tissues. These probes appear to target several, if not all species within the genus, because they labeled all isolates of Hematodinium tested in this study, whereas they were not hybridizing to other parasite species. The PCR-RFLP assay will be invaluable for future studies investigating parasite prevalence, the existence of secondary hosts or environmental reservoirs, and modes of transmission, whereas the DNA probes will be useful for confirming and localizing Hematodinium parasites in crustacean tissues.
Diseases of Aquatic Organisms, 2007
Parasitic dinoflagellates in the genus Hematodinium infect a number of decapod crustaceans in waters off the UK, including the Norway lobster Nephrops norvegicus and the edible crab Cancer pagurus. This study investigated sequence variability in the first internal transcribed spacer (ITS1) region of the ribosomal RNA complex of Hematodinium spp. infecting N. norvegicus, C. pagurus, and Pagurus bernhardus from 4 locations in the UK and from the Hematodinium sp. infecting Chionoecetes opilio from the province of Newfoundland and Labrador, Canada. Phylogenetic analysis of the Hematodinium ITS1 sequences from N. norvegicus, C. pagurus, P. bernhardus and C. opilio suggest that these crustaceans are infected with the same species of Hematodinium. Length variability of the ITS1 region was observed (324 to 345 bp) and attributed to 4 variable microsatellite regions (CATG) n , (GCC) n TCCGC(TG) n , (TA) n , and (GAA) n (GGA) n within the sequenced ITS1 fragment. The observed variation may be due to co-infection of the host crustacean with several different strains of Hematodinium or differences among copies of ITS1 region within the genome of a single parasite cell. The Hematodinium ITS1 sequence from N. norvegicus, C. pagurus, P. bernhardus and C. opilio isolates was sufficiently conserved in primer binding regions targeted by previous molecular diagnostic assays; therefore, we suggest that this assay could be used to screen for Hematodinium infections in these crustacean hosts.
Diseases of Aquatic Organisms, 1995
The amount of DNA recovered from 1 X 106 cells of Hematodinium-like dinoflagellates from Chionoecetes bairdi was 75 ng. PCR (polymerase chain reaction) amplification products were created from the host and Hematodinium spp. Amplified products of 680 base pairs (bp) were produced from Hematodinium-like dinoflagellates from C, bairdi for DNA concentrations of 100, 50, 20, 10 and 1 ng pl-' This amplified fragment was produced only from host tlssue infected with the parasite. The lowest detectable limit of 1 ng of DNA from Hematodinium-like sp. from C. bairdi corresponded to 1.0 X 104 cells. Diagnostic bands of 680 bp were produced from Hematodinjum spp. and Hematodinium-like spp. isolated from the following hosts: Chionoecetes op~lio, Callinectes sapidus, Nephrops norvegicus and Portunus pelagicus. Host DNA (C. bairdi) produced a faint band of 1300 bp for concentrations of 100, 50 and 20 ng pl-'. KEY WORDS: PCR. Dinoflagellate Hematodinium. Decapod et al. 1987, Meyers 1990) and 'watery flesh' in N. norand November 1993 from the Rappalamoda River, vegicus (Field et al. 1992); both of these conditions Chesapeake Bay, Virginia, USA. Tanner crabs Chiomake their flesh unmarketable. noecetes bairdi and C. opilio were collected in Febru-Gross signs of high intensity Hematodinium spp.
Diseases of Aquatic Organisms, 2002
The 18S rRNA gene from Hematodinum sp., a parasitic dinoflagellate that infects blue crabs, was amplified, cloned, and sequenced. The sequence showed a high similarity (95% at the nucleotide level) to sequences obtained from other dinoflagellate species, including both free-living and symbiotic species. Sequence similarity was much lower when compared with parasites of other marine invertebrates with similar life histories and with the 18S rRNA gene from the blue crab. Based on comparison of sequence alignments between Hematodinium, other dinoflagellate species, protozoan pathogens of oysters, and blue crab 18S rRNA gene sequences, 2 sets of PCR primers that specifically amplified fragments of the Hematodinium 18S rRNA gene were developed and tested. One of these primer sets (Hemat-F-1487 and Hemat-R-1654) amplified a 187 bp fragment that could be used routinely as a diagnostic test for the presence of Hematodinium in hemolymph from blue crabs. This fragment was consistently amplified from genomic DNA extracted from hemolymph of Hematodinium infected blue crabs. Comparison between the PCR technique and standard histological examination indicated that the PCR technique was reliable and provided 1000 times more sensitivity than the histological methods. The sensitivity of the PCR diagnostic was estimated to be one parasite cell among 300 000 crab hemocytes. Preliminary studies using the PCR diagnostic technique suggest that Hematodinium sp. is absent in crabs collected from waters with low salinity (5 to 10 ppt), but common in crabs from higher salinity environments in estuarine waters from southeastern Georgia (USA).
Diseases of Aquatic Organisms, 1994
In coastal bays of Maryland and Virginia, USA, adult and juvenile blue crabs Callinectes sapidus were severely infected with the parasitic dinoflagellate I-lematodinium perezi. Dinoflagellates were observed in the hemocoel of all infected crabs; associated histopathological changes were evident in some tissues. Dinoflagellates could be observed with an inverted microscope through the 5th pleopod of heavily infected juvenile crabs (5 to 29 mm) without invasion. This note documents a high prevalence of H. perezj infections in juvenile blue crabs from coastal bays in Maryland and Virginia. The seasonal infection prevalence cycle reported by previous authors is consistent with observations made during this study.
Parasites & Vectors
Background: The parasitic dinoflagellates of the genus Hematodinium represent the causative agent of so-called bitter or pink crab disease in a broad range of shellfish taxa. Outbreaks of Hematodinium-associated disease can devastate local fishing and aquaculture efforts. The goal of our study was to examine the potential role of the common shore (green) crab Carcinus maenas as a reservoir for Hematodinium. Carcinus maenas is native to all shores of the UK and Ireland and the North East Atlantic but has been introduced to, and subsequently invaded waters of, the USA, South Africa and Australia. This species is notable for its capacity to harbour a range of micro-and macro-parasites, and therefore may act as a vector for disease transfer.
Diseases of aquatic organisms, 2016
The dinoflagellate Hematodinium perezi is a prolific pathogen of the blue crab Callinectes sapidus along the Atlantic and Gulf of Mexico coasts of North America. High prevalence, sometimes approaching 100%, and outbreaks with high mortality are associated with higher salinities. H. perezi has not been reported previously in blue crabs from Louisiana, USA, where salinities in coastal habitats are generally below the parasite's favorable range. However, the possibility that H. perezi infects blue crabs in higher salinity habitats offshore has not been investigated. A PCR-based test for H. perezi was used to screen blue crabs collected from both high and low salinity areas. These included juvenile and adult crabs from inshore marshes where salinities are relatively low and from higher salinity offshore shoals that are spawning sites for females. H. perezi was detected in blue crabs from offshore shoals (prevalence = 5.6%) but not in juvenile or adult crabs from inshore habitats. Me...
Evidence for a free-living life stage of the blue crab parasitic dinoflagelate, Hematodinium sp
Harmful Algae, 2006
Hematodinium sp. is a parasitic dinoflagellate reported to cause disease and death in a variety of crustacean species including the blue crab (Callinectes sapidus). However, because of difficulties in the culture of Hematodinium sp. associated with blue crabs, little is known about its life cycle or mode of transmission. Here, we report the first detection of this organism outside of a metazoan host and provide evidence that this life stage can act as an infective agent. Observations of dinospores in crab hemolymph samples suggest that dinospores may be responsible for waterborne disease transmission. Additionally, we developed and validated a quantitative Real Time PCR assay for the detection of Hematodinium sp. inside and outside of a host organism that will be useful for future investigations of Hematodinium biology and Hematodinium sp.-infection etiology. Based on the observations of a free-living form of Hematodinium sp. and the association of this parasite with a widespread epizootic in blue crab populations, we propose that Hematodinium sp. be considered a Harmful Algal Bloom species. # 2005 Published by Elsevier B.V.
Diseases of Aquatic Organisms, 2009
Hematodinium sp. is a parasitic dinoflagellate infecting the blue crab Callinectes sapidus and other crustaceans. PCR-based assays are currently being used to identify infections in crabs that would have been undetectable by traditional microscopic examination. We therefore sought to define the limits of quantitative PCR (qPCR) detection within the context of field collection protocols. We present a qPCR assay based on the Hematodinium sp. 18S rRNA gene that can detect 10 copies of the gene per reaction. Analysis of a cell dilution series vs. defined numbers of a cloned Hematodinium sp. 18S rRNA gene suggests a copy number of 10 000 per parasite and predicts a sensitivity of 0.001 cell equivalents. In practice, the assays are based on analysis of 1% of the DNA extracted from 200 µl of serum, yielding a theoretical detection limit of 5 cells ml -1 hemolymph, assuming that 1 cell is present per sample. When applied to a limited field survey of blue crabs collected in Maryland coastal bays from May to August 2005, 24 of 128 crabs (18.8%) were identified as positive for Hematodinium sp. infection using qPCR. In comparison, only 6 of 128 crabs (4.7%) were identified as positive using traditional hemolymph microscopic examination. The qPCR method also detected the parasite in gill, muscle, heart and hepatopancreas tissues, with 17.2% of the crabs showing infection in at least one of these tissues. Importantly, it is now possible to enumerate parasites within defined quantities of crab tissue, which permits collection of more detailed information on the epizootiology of the pathogen.