DOCSLIB.ORG

Molecular Detection of Hematodinium Spp. in Norway Lobster Nephrops Norvegicus and Other Crustaceans

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Molecular Detection of Hematodinium Spp. in Norway Lobster Nephrops Norvegicus and Other Crustaceans DISEASES OF AQUATIC ORGANISMS Vol. 69: 185–195, 2006 Published April 6 Dis Aquat Org Molecular detection of Hematodinium spp. in Norway lobster Nephrops norvegicus and other crustaceans H. J. Small1, 4,*, D. M. Neil1, A. C. Taylor1, R. J. A. Atkinson2, G. H. Coombs3 1Division of Environmental and Evolutionary Biology, and 3Division of Infection & Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK 2University Marine Biological Association, Millport, Isle of Cumbrae KA28 0EG, UK 4Present address: Virginia Institute of Marine Science, College of William and Mary, Gloucester Point, Virginia 23062, USA ABSTRACT: The Norway lobster Nephrops norvegicus (L.) from the coastal waters of Scotland is seasonally infected by a parasitic dinoflagellate of the genus Hematodinium. Methods used to detect infection include a morphological index (pleopod diagnosis) and several immunoassays. The present study describes the development and application of a set of Hematodinium-specific polymerase chain reaction (PCR) primers and DNA probes based on Hematodinium ribosomal DNA (rDNA). In the PCR assay, a diagnostic band of 380 bp was consistently amplified from total genomic DNA isolated from Hematodinium-infected N. norvegicus. The sensitivity of the assay was 1 ng DNA, which is equivalent to 0.6 parasites. The primer pair also detected Hematodinium DNA in preparations of the amphipod Orchomene nanus, indicating that the amphipod may be infected with the same Hemato- dinium sp. infecting N. norvegicus. DNA probes detected Hematodinium parasites in heart, hepatopancreas and gill tissues from N. norvegicus, and hepatopancreas and gill tissues from Carcinus maenas, confirming Hematodinium infection in the latter. KEY WORDS: Hematodinium · Parasite · Norway lobster · Molecular detection Resale or republication not permitted without written consent of the publisher INTRODUCTION 1996), Ovalipes ocellatus (MacLean & Ruddell 1978) and Portunus pelagicus (Hudson & Shields 1994). In Infection of the Norway lobster Nephrops norvegi- each of the above examples, infection by Hemato- cus by a parasitic dinoflagellate of the genus Hemato- dinium species caused serious mortalities in the host dinium has been described from a number of locations decapod population. around the West Coast of Scotland and the Irish Sea Previous methods to diagnose infection of Nephrops (Field et al. 1992, 1998, Briggs & McAliskey 1996, 2002, norvegicus by Hematodinium species include an Appleton et al. 1997). In UK waters, the edible crab assessment of the external colouration of the carapace Cancer pagurus has also recently been found to har- and appendages of infected lobsters. However this bour Hematodinium spp. (Stentiford et al. 2002). Para- diagnostic method lacks sensitivity and only advanced sitic Hematodinium species have previously been infections can be reliably identified (Stentiford et al. reported infecting several decapod crustaceans includ- 2001a). The pleopods of infected lobsters can be exam- ing Callinectes sapidus (Newman & Johnson 1975, ined under low power light microscopy for the aggre- Messick 1994, Messick & Shields 2000), Cancer pagu- gation of parasites in the vasculature (Field & Appleton rus (Latrouite et al. 1988), Chionoecetes bairdi (Meyers 1995). The severity of infection was classified on a et al. 1987, Love et al. 1993), Chionoecetes opilio (Tay- 5-point scale, from apparently uninfected to an ad- lor & Khan 1995), Necora puber (Wilhelm & Mialhe vanced infection. The pleopod method is reliable as a *Email: [email protected] © Inter-Research 2006 · www.int-res.com 186 Dis Aquat Org 69: 185–195, 2006 field method for identifying advanced infections, but and uninfected haemolymph samples were retained cannot detect tissue-based and low-level haemolymph for DNA isolation. For lobsters having a low-level infection. Immunological methods including an indi- infection (as indicated by ELISA), a subsample of the rect fluorescent antibody technique (IFAT), Western same haemolymph was used to estimate parasite cell blot and enzyme linked immunosorbent assay (ELISA) numbers (ml haemolymph–1) using an improved Neu- have all been developed using a polyclonal rabbit anti- bauer counting chamber. serum raised against a mixed in vitro culture of vegeta- Tissue samples from infected and uninfected lobsters tive forms of Hematodinium from N. norvegicus (Field were fixed in Davidson’s seawater fixative (20 ml & Appleton 1996, Stentiford et al. 2001b, Small et al. formalin [40% v/v], 10 ml glycerol, 10 ml glacial acetic 2002). However, the polyclonal antibody cross-reacts acid, 30 ml 100% ethanol, 30 ml seawater) for 24 h, with epitopes found on other protozoan parasites dehydrated in an ethanol series and embedded in (Bushek et al. 2002) and may not recognise in vivo life paraffin wax. Several other samples of haemolymph cycle stages of Hematodinium species that were ab- and tissue from crustacean and dinoflagellate species sent from the original innoculum or present only in commonly found in UK waters were also retained and other hosts. Because of the above concerns and the prepared for DNA extraction (see Table 1). Davidson’s need for more sensitive and specific diagnostic fixed-tissue sections from a shore crab (Carcinus methods, a molecular approach to parasite detection maenas) collected from the English Channel with was undertaken. a probable Hematodinium infection were obtained DNA-based diagnostic methods utilising the poly- from Dr. G. Stentiford (CEFAS Weymouth Laboratory). merase chain reaction (PCR) and in situ hybridisation H. perezi cells from in vitro culture (Small 2004) were (ISH) assays have facilitated diagnosis of many marine preserved in 100% ethanol prior to DNA extraction. pathogens of shellfish (for review see Cunningham To amplify parasite DNA for oligonucleotide primer 2002). Nuclear ribosomal DNA (rDNA) is widely design, 1 × 105 parasites from an in vitro culture of acknowledged as a useful target for the definition of Hematodinium sp. isolated from the Norway lobster genetic markers informative at several levels (Gasser & (Appleton & Vickerman 1998) were collected by centri- Zhu 1999). The rate of evolution varies between differ- fugation (1000 × g for 4 min at 4°C), the resultant super- ent regions of rDNA, resulting in target sequences natant was removed, and the sedimented cells pro- ranging from highly conserved to highly variable cessed for DNA extraction (see next subsection). The (Hillis & Dixon 1991). PCR primers used to diagnose same centrifugation procedure was also carried out for Hematodinium infection in crustacean hosts have pre- the free-living dinoflagellates Alexandrium tamarense viously been reported (Hudson & Adlard 1994, Gruebl (Culture Collection of Algae and Protozoa, CCAP et al. 2002). However, both primer sets used in those 1119/5), Gymnodinium catenatum (CCAP 1117/6) studies were located in the conserved 18S and 5.8S (both 1 × 105 cells), H. perezi and a Mesanophrys-like regions of the rDNA complex and are not specific for parasitic ciliate (5 × 104 cells) prior to DNA extraction. individual Hematodinium species. Variable regions of To examine a possible secondary host for the Hema- rDNA from other dinoflagellates have been identified todinium sp. infecting Nephrops norvegicus, amphi- allowing species-specific PCR assays to be developed pods Orchomene nanus were captured in baited traps (Litaker et al. 2003). We describe herein the develop- from the Hunterston Channel in the Clyde Sea area in ment and application of PCR and ISH assays for detec- August 2001, March 2003 and August 2003, according tion of Hematodinium species infection in Nephrops to the method of Moore & Wong (1995). Previous stud- norvegicus and other crustaceans. ies had indicated that there was a high prevalence of Hematodinium sp. infection of Norway lobsters at this location (J. Atkinson pers. comm.). Individual O. nanus MATERIALS AND METHODS were held for 3 d in seawater at 12°C prior to preserva- tion in 100% ethanol or Davidson’s seawater fixative Sample collection and preparation. Norway lobsters (only the March 2003 and August 2003 samples). Nephrops norvegicus and other crustaceans used in Genomic DNA samples were prepared from each this study (unless indicated otherwise) were caught by amphipod and assayed by PCR for the presence of otter bottom-trawl (70 mm mesh size) south of Little Hematodinium spp. Cumbrae in the Clyde Sea Area, Scotland, UK. DNA extraction. Genomic DNA was extracted from Haemolymph samples were taken from the base of the 100 µl haemolymph samples, 100 mg tissue samples, fifth pereiopod, using a sterile 1 ml disposable syringe the in vitro Hematodinium sp., ciliate and dinoflagel- and 25-gauge needle, and were frozen at –20°C. Sam- late pellets, and from individual whole Orchomene ples were assayed for the presence of Hematodinium nanus according to standard procedures (Sambrook by the ELISA method of Small et al. (2002). Infected et al. 1989). Briefly, samples were homogenised/ Small et al.: Molecular detection of Hematodinium spp. 187 resuspended in 250 µl extraction buffer (50 mM Tris, software/BOX_form.html). Sequences were compared 5 mM EDTA, 100 mM NaCl, pH 8), 100 µl of 10% for similarity to those of other dinoflagellates by Basic SDS (w/v) and Proteinase K (0.28 ng µl–1) and incu- Local Alignment Tool searches (BLAST; Altschul et al. bated at 56°C for 18 to 24 h. All DNAs were purified 1990) in GenBank. Suitable priming regions for PCR by single-step standard phenol/chloroform
Load more

Recommended publications
  • An Aerobic Eukaryotic Parasite with Functional Mitochondria That Likely
    An aerobic eukaryotic parasite with functional mitochondria that likely lacks a mitochondrial genome Uwe John, Yameng Lu, Sylke Wohlrab, Marco Groth, Jan Janouškovec, Gurjeet Kohli, Felix Mark, Ulf Bickmeyer, Sarah Farhat, Marius Felder, et al. To cite this version: Uwe John, Yameng Lu, Sylke Wohlrab, Marco Groth, Jan Janouškovec, et al.. An aerobic eukaryotic parasite with functional mitochondria that likely lacks a mitochondrial genome. Science Advances , American Association for the Advancement of Science (AAAS), 2019, 5 (4), pp.eaav1110. 10.1126/sci- adv.aav1110. hal-02372304 HAL Id: hal-02372304 https://hal.archives-ouvertes.fr/hal-02372304 Submitted on 25 Nov 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. SCIENCE ADVANCES | RESEARCH ARTICLE EVOLUTIONARY BIOLOGY Copyright © 2019 The Authors, some rights reserved; An aerobic eukaryotic parasite with functional exclusive licensee American Association mitochondria that likely lacks a mitochondrial genome for the Advancement Uwe John1,2*, Yameng Lu1,3, Sylke Wohlrab1,2, Marco Groth4, Jan Janouškovec5, Gurjeet S. Kohli1,6, of Science. No claim to 1 1 7 4 1,8 original U.S. Government Felix C. Mark , Ulf Bickmeyer , Sarah Farhat , Marius Felder , Stephan Frickenhaus , Works.
    [Show full text]
  • Molecular Data and the Evolutionary History of Dinoflagellates by Juan Fernando Saldarriaga Echavarria Diplom, Ruprecht-Karls-Un
    Molecular data and the evolutionary history of dinoflagellates by Juan Fernando Saldarriaga Echavarria Diplom, Ruprecht-Karls-Universitat Heidelberg, 1993 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE STUDIES Department of Botany We accept this thesis as conforming to the required standard THE UNIVERSITY OF BRITISH COLUMBIA November 2003 © Juan Fernando Saldarriaga Echavarria, 2003 ABSTRACT New sequences of ribosomal and protein genes were combined with available morphological and paleontological data to produce a phylogenetic framework for dinoflagellates. The evolutionary history of some of the major morphological features of the group was then investigated in the light of that framework. Phylogenetic trees of dinoflagellates based on the small subunit ribosomal RNA gene (SSU) are generally poorly resolved but include many well- supported clades, and while combined analyses of SSU and LSU (large subunit ribosomal RNA) improve the support for several nodes, they are still generally unsatisfactory. Protein-gene based trees lack the degree of species representation necessary for meaningful in-group phylogenetic analyses, but do provide important insights to the phylogenetic position of dinoflagellates as a whole and on the identity of their close relatives. Molecular data agree with paleontology in suggesting an early evolutionary radiation of the group, but whereas paleontological data include only taxa with fossilizable cysts, the new data examined here establish that this radiation event included all dinokaryotic lineages, including athecate forms. Plastids were lost and replaced many times in dinoflagellates, a situation entirely unique for this group. Histones could well have been lost earlier in the lineage than previously assumed.
    [Show full text]
  • The Mitochondrial Genome and Transcriptome of the Basal
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE GBEprovided by PubMed Central The Mitochondrial Genome and Transcriptome of the Basal Dinoflagellate Hematodinium sp.: Character Evolution within the Highly Derived Mitochondrial Genomes of Dinoflagellates C. J. Jackson, S. G. Gornik, and R. F. Waller* School of Botany, University of Melbourne, Australia *Corresponding author: E-mail: [email protected]. Accepted: 12 November 2011 Abstract 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.
    [Show full text]
  • <I>Hematodinium Perezi</I>
    Old Dominion University ODU Digital Commons Biological Sciences Faculty Publications Biological Sciences 2019 Parasitic Dinoflagellate Hematodinium perezi Prevalence in Larval and Juvenile Blue Crabs Callinectes sapidus from Coastal Bays of Virginia H. J. Small J. P. Huchin-Mian K. S. Reece K. M. Pagenkopp Lohan Mark J. Butler IV Old Dominion University, [email protected] See next page for additional authors Follow this and additional works at: https://digitalcommons.odu.edu/biology_fac_pubs Part of the Biology Commons, Marine Biology Commons, and the Parasitology Commons Original Publication Citation Small, H. J., Huchin-Mian, J. P., Reece, K. S., Lohan, K. M. P., Butler, M. J., & Shields, J. D. (2019). Parasitic dinoflagellate Hematodinium perezi prevalence in larval and juvenile blue crabs Callinectes sapidus from coastal bays of Virginia. Diseases of Aquatic Organisms, 134(3), 215-222. doi:10.3354/dao03371 This Article is brought to you for free and open access by the Biological Sciences at ODU Digital Commons. It has been accepted for inclusion in Biological Sciences Faculty Publications by an authorized administrator of ODU Digital Commons. For more information, please contact [email protected]. Authors H. J. Small, J. P. Huchin-Mian, K. S. Reece, K. M. Pagenkopp Lohan, Mark J. Butler IV, and J. D. Shields This article is available at ODU Digital Commons: https://digitalcommons.odu.edu/biology_fac_pubs/391 Vol. 134: 215–222, 2019 DISEASES OF AQUATIC ORGANISMS Published online June 6 https://doi.org/10.3354/dao03371 Dis Aquat Org OPENPEN ACCESSCCESS Parasitic dinoflagellate Hematodinium perezi prevalence in larval and juvenile blue crabs Callinectes sapidus from coastal bays of Virginia H.
    [Show full text]
  • Environmental DNA Metabarcoding for Simultaneous Monitoring and Ecological Assessment of Many Harmful Algae
    bioRxiv preprint doi: https://doi.org/10.1101/2020.10.01.322941; this version posted October 2, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Environmental DNA Metabarcoding for Simultaneous Monitoring and Ecological Assessment of Many Harmful Algae Emily Jacobs-Palmer1, Ramón Gallego1,2, Kelly Cribari1, Abigail Keller1, Ryan P. Kelly1 1University of Washington School of Marine and Environmental Affairs 2NRC Research Associateship Program, Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration. Abstract Harmful algae can have profound economic, environmental, and social consequences. As the timing, frequency, and severity of harmful algal blooms (HABs) change alongside global climate, efficient tools to monitor and understand the current ecological context of these taxa are increasingly important. Here we employ environmental DNA metabarcoding to identify patterns in a wide variety of harmful algae and associated ecological communities in the Hood Canal of Puget Sound in Washington State, USA. We track trends of presence and abundance in a series of water samples across nearly two years. We find putative harmful algal sequences in a majority of samples, suggesting that these groups are routinely present in local waters. We report patterns in variants of the economically important genus Pseudo-nitzschia (family Bacillariaceae), as well as multiple harmful algal taxa previously unknown or poorly documented in the region, including a cold-water variant from the saxitoxin-producing genus Alexandrium (family Gonyaulacaceae), two variants from the karlotoxin-producing genus Karlodinium (family Kareniaceae), and one variant from the parasitic genus Hematodinium (family Syndiniaceae).
    [Show full text]
  • 11 Shields FISH 98(1)
    139 Abstract.–On the eastern seaboard of Mortality and hematology of blue crabs, the United States, populations of the blue crab, Callinectes sapidus, experi- Callinectes sapidus, experimentally infected ence recurring outbreaks of a parasitic dinoflagellate, Hematodinium perezi. with the parasitic dinoflagellate Epizootics fulminate in summer and autumn causing mortalities in high- Hematodinium perezi* salinity embayments and estuaries. In laboratory studies, we experimentally investigated host mortality due to the Jeffrey D. Shields disease, assessed differential hemato- Christopher M. Squyars logical changes in infected crabs, and Department of Environmental Sciences examined proliferation of the parasite. Virginia Institute of Marine Science Mature, overwintering, nonovigerous The College of William and Mary female crabs were injected with 103 or P.O. Box 1346, Gloucester Point, VA 23602, USA 105 cells of H. perezi. Mortalities began E-mail address (for J. D. Shields): [email protected] 14 d after infection, with a median time to death of 30.3 ±1.5 d (SE). Sub- sequent mortality rates were greater than 86% in infected crabs. A relative risk model indicated that infected crabs were seven to eight times more likely to Hematodinium perezi is a parasitic larger, riverine (“bayside”) fishery; die than controls and that decreases in total hemocyte densities covaried signif- dinoflagellate that proliferates in it appears most detrimental to the icantly with mortality. Hemocyte densi- the hemolymph of several crab spe- coastal (“seaside”) crab fisheries. ties declined precipitously (mean=48%) cies. In the blue crab, Callinectes Outbreaks of infestation by Hema- within 3 d of infection and exhibited sapidus, H. perezi is highly patho- todinium spp.
    [Show full text]
  • Molecular Phylogeny and Surface Morphology of Colpodella Edax (Alveolata): Insights Into the Phagotrophic Ancestry of Apicomplexans
    J. Eukaryot. Microbiol., 50(5), 2003 pp. 334±340 q 2003 by the Society of Protozoologists Molecular Phylogeny and Surface Morphology of Colpodella edax (Alveolata): Insights into the Phagotrophic Ancestry of Apicomplexans BRIAN S. LEANDER,a OLGA N. KUVARDINA,b VLADIMIR V. ALESHIN,b ALEXANDER P. MYLNIKOVc and PATRICK J. KEELINGa aCanadian Institute for Advanced Research, Program in Evolutionary Biology, Department of Botany, University of British Columbia, Vancouver, BC, V6T 1Z4, Canada, and bDepartments of Evolutionary Biochemistry and Invertebrate Zoology, Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, 119 992, Russian Federation, and cInstitute for the Biology of Inland Waters, Russian Academy of Sciences, Borok, Yaroslavskaya oblast, 152742, Russian Federation ABSTRACT. The molecular phylogeny of colpodellids provides a framework for inferences about the earliest stages in apicomplexan evolution and the characteristics of the last common ancestor of apicomplexans and dino¯agellates. We extended this research by presenting phylogenetic analyses of small subunit rRNA gene sequences from Colpodella edax and three unidenti®ed eukaryotes published from molecular phylogenetic surveys of anoxic environments. Phylogenetic analyses consistently showed C. edax and the environmental sequences nested within a colpodellid clade, which formed the sister group to (eu)apicomplexans. We also presented surface details of C. edax using scanning electron microscopy in order to supplement previous ultrastructural investigations of this species using transmission electron microscopy and to provide morphological context for interpreting environmental sequences. The microscopical data con®rmed a sparse distribution of micropores, an amphiesma consisting of small polygonal alveoli, ¯agellar hairs on the anterior ¯agellum, and a rostrum molded by the underlying (open-sided) conoid.
    [Show full text]
  • Infection by a Hematodinium-Like Parasitic Dinoflagellate Causes Pink
    Journal of Invertebrate Pathology 79 (2002) 179–191 www.academicpress.com Infection by a Hematodinium-like parasitic dinoflagellate causes Pink Crab Disease (PCD) in the edible crab Cancer pagurus G.D. Stentiford,a,* M. Green,a K. Bateman,a H.J. Small,b D.M. Neil,b and S.W. Feista a Centre for Environment, Fisheries and Aquaculture Science (CEFAS) Weymouth Laboratory, Barrack Road, The Nothe, Weymouth, Dorset DT4 8UB, UK b Division of Environmental and Evolutionary Biology, Graham Kerr Building, University of Glasgow, Glasgow G12 8QQ, Scotland, UK Received 30 October 2001; accepted 2 May 2002 Abstract The edible crab (Cancer pagurus) supports a large and valuable fishery in UK waters. Much of the catch is transported live to continental Europe in specially designed live-well (‘vivier’) vehicles. During the winter of 2000/2001, many trap-caught crabs from Guernsey, Channel Islands, UK, were reportedly moribund and pink in colour. These crabs generally died before and during vivier transportation. We provide histological, immunological, and molecular evidence that this condition is associated with infection by a Hematodinium-like dinoflagellate parasite similar to that previously reported in C. pagurus and to an infection causing seasonal mass mortalities of the Norway lobster (Nephrops norvegicus). Pathologically, every altered host bore the infection, which was charac- terised by very large numbers of plasmodial and vegetative stages in the haemolymph and depletion of reserve cells in the he- patopancreas. Due to the hyperpigmentation of the carapace and appendages, we have called this infection ‘Pink Crab Disease’ (PCD). Similar Hematodinium infections cause ‘Bitter Crab Disease’ in tanner and snow crabs, which has had a negative effect on their marketability.
    [Show full text]
  • Hematodinium Sp
    crustacean Protozoa Hematodinium sp. - Bitter Crab Disease of Tanner Crabs I. Causative Agent and Disease IV. Transmission Bitter crab disease is caused by a Although the natural route of trans- parasitic dinoflagellate,Hematodinium mission has not been established, there sp. belonging to the protozoan phylum are likely to be several modes in which Alveolata, subphylum Dinoflagellida, the parasite is transmitted horizontally order Syndinida and family Syndiniceae. via: stages of the parasite in seawater A major feature of classification for di- that enter breaks in the cuticle during noflagellates is the biflagellated grooved mating and molting; cannibalism or dinospore stage. Obligate Hematodinium feeding on detritis containing resting parasites have been described from spores; sexual transmission via semi- several species of crustaceans, certain nal fluids of parasitized male crabs and fishes and cephalopods. In crustaceans possibly through a reservoir host such as the type species is H. perezi parasitizing bottom dwelling amphipods. The para- the European shore crab. All Hema- site and disease have been transmitted in todinium described from crustaceans the laboratory by injection of hemo- parasitize the hemolymph causing lymph containing vegetative trophonts systemic disease and mortality affecting and dinospore stages. Life history stages at least 26 species of crustacean hosts in include: trophonts which are single cells Europe, Australia and North America and have slow division; larger plasmo- including many commercially important dia with multiple nuclei; pre-spores crab species and 13 species of benthic with multiple nuclei and rapid division amphipods. followed by sporulation of biflagellated spores with a single nucleus. Spores II. Host Species are of two types, a large macrospore The bitter crab Hematodinium is and a small microspore, generally with reported from Tanner and snow crabs (C.
    [Show full text]
  • Genetic and Phenotypic Diversity Characterization of Natural Populations of the Parasitoid Parvilucifera Sinerae
    Vol. 76: 117–132, 2015 AQUATIC MICROBIAL ECOLOGY Published online October 22 doi: 10.3354/ame01771 Aquat Microb Ecol OPENPEN ACCESSCCESS Genetic and phenotypic diversity characterization of natural populations of the parasitoid Parvilucifera sinerae Marta Turon1, Elisabet Alacid1, Rosa Isabel Figueroa2, Albert Reñé1, Isabel Ferrera1, Isabel Bravo3, Isabel Ramilo3, Esther Garcés1,* 1Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar, CSIC, Pg. Marítim de la Barceloneta 37-49, 08003 Barcelona, Spain 2Department of Biology, Lund University, Box 118, 221 00 Lund, Sweden 3Centro Oceanográfico de Vigo, IEO (Instituto Español de Oceanografía), Subida a Radio Faro 50, 36390 Vigo, Spain ABSTRACT: Parasites exert important top-down control of their host populations. The host−para- site system formed by Alexandrium minutum (Dinophyceae) and Parvilucifera sinerae (Perkinso- zoa) offers an opportunity to advance our knowledge of parasitism in planktonic communities. In this study, DNA extracted from 73 clonal strains of P. sinerae, from 10 different locations along the Atlantic and Mediterranean coasts, was used to genetically characterize this parasitoid at the spe- cies level. All strains showed identical sequences of the small and large subunits and internal tran- scribed spacer of the ribosomal RNA, as well as of the β-tubulin genes. However, the phenotypical characterization showed variability in terms of host invasion, zoospore success, maturation time, half-maximal infection, and infection rate. This characterization grouped the strains within 3 phe- notypic types distinguished by virulence traits. A particular virulence pattern could not be ascribed to host-cell bloom appearance or to the location or year of parasite-strain isolation; rather, some parasitoid strains from the same bloom significantly differed in their virulence traits.
    [Show full text]
  • Molecular Phylogeny and Surface Morphology of Colpodella Edax (Alveolata): Insights Into the Phagotrophic Ancestry of Apicomplexans
    J. Eukaryot. MicroDiol., 50(S), 2003 pp. 334-340 0 2003 by the Society of Protozoologists Molecular Phylogeny and Surface Morphology of Colpodella edax (Alveolata): Insights into the Phagotrophic Ancestry of Apicomplexans BRIAN S. LEANDER,;‘ OLGA N. KUVARDINAP VLADIMIR V. ALESHIN,” ALEXANDER P. MYLNIKOV and PATRICK J. KEELINGa Canadian Institute for Advanced Research, Program in Evolutionary Biology, Departnzent of Botany, University of British Columbia, Vancouver, BC, V6T Iz4, Canada, and hDepartments of Evolutionary Biochemistry and Invertebrate Zoology, Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, I I9 992, Russian Federation, and ‘Institute for the Biology of Inland Waters, Russian Academy qf Sciences, Borok, Yaroslavskaya oblast, I52742, Russian Federation ABSTRACT. The molecular phylogeny of colpodellids provides a framework for inferences about the earliest stages in apicomplexan evolution and the characteristics of the last common ancestor of apicomplexans and dinoflagellates. We extended this research by presenting phylogenetic analyses of small subunit rRNA gene sequences from Colpodella edax and three unidentified eukaryotes published from molecular phylogenetic surveys of anoxic environments. Phylogenetic analyses consistently showed C. edax and the environmental sequences nested within a colpodellid clade, which formed the sister group to (eu)apicomplexans. We also presented surface details of C. edax using scanning electron microscopy in order to supplement previous ultrastructural investigations of this species using transmission electron microscopy and to provide morphological context for interpreting environmental sequences. The microscopical data confirmed a sparse distribution of micropores, an amphiesma consisting of small polygonal alveoli, flagellar hairs on the anterior flagellum, and a rostrum molded by the underlying (open-sided)conoid. Three flagella were present in some individuals, a peculiar feature also found in the microgametes of some apicomplexans.
    [Show full text]
  • Alveolata) Using Small Subunit Rrna Gene Sequences Suggests They Are the Free-Living Sister Group to Apicomplexans
    J. Elrkutyt. Microhiol., 49(6), 2002 pp. 49G.504 0 2002 by the Society of Prutozoolugists The Phylogeny of Colpodellids (Alveolata) Using Small Subunit rRNA Gene Sequences Suggests They are the Free-living Sister Group to Apicomplexans OLGA N. KUVARDINA,’.hBRIAN S. LEANDER,’,aVLADIMIR V. ALESHIN,h ALEXANDER P. MYL’NIKOV,” PATRICK J. KEELING‘‘and TIMUR G. SIMDYANOVh “Canadian Institute for Advunced Resenrch, Program in Evolutionury Biology, Department of Botany, Universiv of British Columbia, Vancouver, BC V6T 124, Canada, and hDepartnzents of Evolutionary Biochemistry and litvertebrate Zoology, Belozersb Institute of Physico-Chemical Biology, Moscow State University, Moscow 119 899, Russian Federation, and ‘Instinrtefor the Biology of Inland Waters, Russian Academy of Sciences, Borok, Yaroslnvskaya oblavt 152742, Russian Federation ABSTRACT. In an attempt to reconstruct early alveolate evolution, we have examined the phylogenetic position of colpodellids by analyzing small subunit rDNA sequences from Colpodella pontica Myl’nikov 2000 and Colpodella sp. (American Type Culture Col- lection 50594). All phylogenetic analyses grouped the colpodellid sequences together with strong support and placed them strongly within the Alveolata. Most analyses showed colpodellids as the sister group to an apicomplexan clade, albeit with weak support. Sequences from two perkinsids, Perkinsus and Parvilucifera, clustered together and consistently branched as the sister group to dino- flagellates as shown previously. These data demonstrate that colpodellids and perkinsids are plesiomorphically similar in morphology and help provide a phylogenetic framework for inferring the combination of character states present in the last common ancestor of dinoflagellates and apicomplexans. We can infer that this ancestor was probably a myzocytotic predator with two heterodynamic flagella, micropores, trichocysts, rhoptries, micronemes, a polar ring, and a coiled open-sided conoid.
    [Show full text]