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1.
Dis Aquat Organ ; 136(1): 89-105, 2019 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-31575837

RESUMO

This study provides morphological, ultrastructural and phylogenetic characterization of 2 novel species of Haplosporidia (Haplosporidium echinogammari n. sp. and H. orchestiae n. sp.) infecting amphipods of the genera Echinogammarus and Orchestia collected in southwestern England. Both parasites infect the connective tissues associated with the digestive gland and the tegument, and eventually infect other organs causing disruption of host tissues with associated motor impairment and fitness reduction. Prevalence of infection varied with host species, provenance and season, being as high as 75% for individuals of E. marinus infected with H. echinogammari in June (n = 50). Although no spores were found in any of the infected amphipods examined (n = 82), the morphology of monokaryotic and dikaryotic unicellular stages of the parasites enabled differentiation between the 2 new species. Phylogenetic analysis of the new species based on the small subunit (SSU) rDNA gene placed H. echinogammari close to H. diporeiae in haplosporidian lineage C, and H. orchestiae in a novel branch within Haplosporidium. Genetic diversity of the haplosporidians infecting these and other amphipod species was evaluated and compared to morphological and ultrastructural changes to host tissues. The phylogenetic relationship of haplosporidian infections in other crustacean hosts is discussed after inclusion into the analysis of 25 novel SSU rDNA sequences obtained from crabs, isopods and crayfish.


Assuntos
Anfípodes/parasitologia , Haplosporídios/classificação , Filogenia , Animais , Inglaterra , Haplosporídios/ultraestrutura
2.
Dis Aquat Organ ; 133(1): 57-68, 2019 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-31089003

RESUMO

The edible mussel Mytilus edulis is a major aquaculture commodity in Europe, with 168000 t produced in 2015. A number of abundant, well characterised parasites of the species are known, though none are considered to cause significant mortality. Haplosporida (Rhizaria, Endomyxa) is an order of protistan parasites of aquatic invertebrates, the best studied of which are the oyster pathogens Haplosporidium nelsoni and Bonamia ostreae. While these species are well characterised within their hosts, the diversity, life-cycle and modes of transmission of haplosporidians are very poorly understood. Haplosporidian parasites have previously been reported from Mytilus spp., however the majority of these remain uncharacterised, and no molecular data exist for any species. In this study, we identified 2 novel haplosporidian parasites of M. edulis present in the UK. The first of these, observed by light microscopy and in situ hybridisation infecting the gills, mantle, gonadal tubules and digestive connective tissues of mussels in the Tamar estuary, England, we describe as Minchinia mytili on the basis of 18S sequence data. The second, observed infecting a single archive specimen collected in Loch Spelve, Mull, Scotland, infects the foot muscle, gills and connective tissue of the digestive gland. Sequence data places this parasite in an uncharacterised clade of sequences amplified from tropical bivalve guts and water samples, sister to H. nelsoni. Screening of water and sediment samples collected at the sample site in the Tamar estuary revealed the presence of both sequence types in the water column, suggesting host-free or planktonic life stages.


Assuntos
Haplosporídios , Animais , Inglaterra , Europa (Continente) , Escócia
3.
PLoS One ; 14(2): e0212028, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30794588

RESUMO

Noble pen shell or fan mussel, Pinna nobilis Linnaeus (1758), protected since 1992, was incorporated into the Spanish Catalogue of Threatened Species (Category: Vulnerable, Royal Decree 139/2011). The status is presently in the process of being catalogued as critically endangered, pending approval by Spanish Government (https://www.mapama.gob.es/es/biodiversidad/participacion-publica/Borrador_OM_situacion_critica.aspx). The International Union for the Conservation of Nature (IUCN) alerted the countries of the Mediterranean basin to the "emergent situation" due to serious mortality events suffered by the fan mussel, putting it in serious risk of extinction. Thus, emergency actions have been implemented by Spanish authorities in which several research institutes from all over the country are involved. The parasite, Haplosporidium pinnae, was recently characterized by histology, TEM, SEM and molecular biology techniques and it was considered responsible for the mass mortality of P. nobilis in the Mediterranean Sea. In this context, the aim of this study has been to develop species-specific quantitative PCR (qPCR) protocol carrying out a fast, specific and effective molecular diagnose of H. pinnae. In this sense, the detection limit for qPCR was equal to 30 copies of SSU rDNA / ng of DNA using plasmid alone and when 100ng DNA of non-infected oyster were added. The qPCR assay revealed that 94% of the 32 analysed mantle tissues of fan mussel were infected by H. pinnae, showing a high sensitivity and specificity for its detection (100% if we don't consider negative and too much degraded samples). This technique will allow us to make quicker follow-ups of the disease, allowing us to get a better understanding of its evolution in order to help in the rescue of P. nobilis populations.


Assuntos
Bivalves/parasitologia , Haplosporídios/isolamento & purificação , Infecções Protozoárias em Animais/diagnóstico , Animais , DNA Ribossômico/genética , Haplosporídios/genética , Região do Mediterrâneo , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Análise de Sequência de DNA , Especificidade da Espécie
4.
J Fish Dis ; 42(1): 63-74, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30324720

RESUMO

Bonamia spp. parasites threaten flat oyster (Ostrea spp.) farming worldwide. Understanding test performance is important for designing surveillance and interpreting diagnostic results. Following a pilot survey which found low Bonamia sp. intensity in farmed Ostrea angasi, we tested further oysters (n = 100-150) from each of three farms for Bonamia sp. using heart smear, histology and qPCR. We used a Bayesian Latent Class Model to assess diagnostic sensitivity (DSe) and specificity (DSp) of these tests individually or in combination, and to assess prevalence. Histology was the best individual test (DSe 0.76, DSp 0.93) compared to quantitative polymerase chain reaction (qPCR) (DSe 0.69, DSp 0.93) and heart smear (DSe 0.61, DSp 0.60). Histology combined with qPCR and defining a positive from either test as an infected case maximized test performance (DSe 0.91, DSp 0.88). Prevalence was higher at two farms in a high-density oyster growing region than at a farm cultivating oysters at lower density. Parasite intensities were lower than in New Zealand and European studies, and this is probably contributed to differences in the performance of test when compared to other studies. Understanding diagnostic test performance in different populations can support the development of improved Bonamia surveillance programs.


Assuntos
Doenças dos Peixes/diagnóstico , Doenças dos Peixes/parasitologia , Haplosporídios , Ostrea/parasitologia , Infecções Protozoárias em Animais/diagnóstico , Animais , Aquicultura , Doenças dos Peixes/epidemiologia , Coração/parasitologia , Técnicas Histológicas/veterinária , Prevalência , Infecções Protozoárias em Animais/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real , Austrália do Sul/epidemiologia
5.
J Invertebr Pathol ; 158: 6-15, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30130536

RESUMO

The purpose of this study was to compare the parasites of two coexisting bivalves, the edible Puelche oyster (Ostrea puelchana) and the false oyster (Pododesmus rudis) that lives attached to O. puelchana shells, and to investigate their host specificity. Samples from wild populations, 465 O. puelchana and 131 P. rudis, were collected seasonally during two years in the San José Gulf (northern Patagonia, Argentina) and were processed using standard histological techniques. To increase the natural low prevalences of Bonamia spp. and Perkinsus spp. that are present in wild populations, an in situ experiment was performed by maintaining captive sentinel bivalves at high densities inside a plastic mesh bag to enhance parasite transmission. Polymerase chain reaction (PCR) assays were used to test for apparent Bonamia sp. infections among captive sentinel O. puelchana specimens (n = 80), and Ray's fluid thioglycollate medium (RFTM) assays and histological immunoassays tested for apparent Perkinsus sp. infections among captive sentinel P. rudis specimens (n = 100). Despite histological observations that revealed the presence of microcells resembling Bonamia sp. infecting hemocytes of some Puelche oysters, PCR assays did not confirm that parasite identification. Among captive sentinel P. rudis that showed histological evidence of Perkinsus sp. infections, neither RFTM nor immunoassays confirmed such parasites. Ostrea puelchana from wild populations were occasional hosts for both Rickettsia-like organism (RLOs) and Urastoma-like turbellarians. In contrast, six parasite taxa infected P. rudis from coexisting populations, including RLOs, Urastoma-like turbellarians, an intracellular gregarine species, Nematopsis-like oocysts, an unidentified coccidian and a Perkinsus qugwadi-like protozoan. These results demonstrated specific infection patterns of the identified parasites in relation to their hosts.


Assuntos
Ostreidae/parasitologia , Frutos do Mar/parasitologia , Animais , Oceano Atlântico , Haplosporídios
6.
Sci Rep ; 8(1): 12494, 2018 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-30131502

RESUMO

Bonamia ostreae has been associated with the decline of flat oyster Ostrea edulis populations in some European countries. This obligatory intracellular parasite persists and multiplies into hemocytes. Previous in vitro experiments showed that apoptosis is activated in hemocytes between 1 h and 4 h of contact with the parasite. The flat oyster uses the apoptosis pathway to defend against B. ostreae. However, the parasite might be also able to modulate this response in order to survive in its host. In order to investigate this hypothesis the apoptotic response of the host was evaluated using flow cytometry, transmission electron microscopy and by measuring the response of genes involved in the apoptotic pathway after 4 h. In parallel, the parasite response was investigated by measuring the expression of B. ostreae genes involved in different biological functions including cell cycle and cell death. Obtained results allow describing molecular apoptotic pathways in O. edulis and confirm that apoptosis is early activated in hemocytes after a contact with B. ostreae. Interestingly, at cellular and molecular levels this process appeared downregulated after 44 h of contact. Concurrently, parasite gene expression appeared reduced suggesting that the parasite could inhibit its own metabolism to escape the immune response.


Assuntos
Perfilação da Expressão Gênica/veterinária , Redes Reguladoras de Genes , Haplosporídios/patogenicidade , Ostrea/parasitologia , Animais , Apoptose , Ciclo Celular , Europa (Continente) , Citometria de Fluxo , Regulação da Expressão Gênica , Haplosporídios/genética , Hemócitos/parasitologia , Interações Hospedeiro-Parasita , Microscopia Eletrônica de Transmissão , Ostrea/genética , Análise de Sequência de RNA/veterinária
7.
J Invertebr Pathol ; 157: 9-24, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30005968

RESUMO

This study provides morphological and molecular characterization of a new species, Haplosporidium pinnae), very likely responsible for mass mortality of fan mussels, Pinna nobilis, in the Western Mediterranean Sea. The parasite was found in dead or moribund P. nobilis but did not occur in healthy fan mussels from locations that were not affected by abnormal mortality. Histological examination of infected fan mussels showed uninucleate cells of a haplosporidan parasite throughout the connective tissue and hemolymph sinuses of the visceral mass and binucleate cells and, rarely, multinucleate plasmodia were also detected in the connective tissue. Additionally, stages of sporulation occurred in the epithelium of the host digestive gland tubules. Spores were slightly ellipsoidal with a hinged operculum in one pole. Typical haplosporosomes were not found with TEM but vesicles with two concentric membranes resembling haplosporosomes were abundant in the cytoplasm of the multinucleate plasmodia occurring in host digestive gland tubules. SEM analysis showed multiple structures on the spore surface; some spores had two or four long tape-like filaments attached to the spore wall. Phylogenetic analysis based on the SSU rDNA sequence placed this parasite within a large clade including species of the order Haplosporida, not in the Bonamia/Minchinia subclade or the subclade containing most Haplosporidium species, but within a subclade of Haplosporidium sp. from Penaeus vannamei. Our results suggested that H. pinnae and the parasite of P. vannamei may represent a distinct new genus within the order Haplosporida.


Assuntos
Bivalves/parasitologia , Haplosporídios/genética , Infecções Protozoárias em Animais , Animais , DNA Ribossômico/análise , Genes de Protozoários/genética , Haplosporídios/classificação , Mar Mediterrâneo , Filogenia , Frutos do Mar/parasitologia
8.
Dis Aquat Organ ; 128(2): 127-145, 2018 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-29733027

RESUMO

European flat oyster Ostrea edulis populations have suffered extensive mortalities caused by bonamiosis. The protozoan parasite Bonamia ostreae is largely responsible for this disease in Europe, while its congener B. exitiosa has been detected more recently in various European countries. Both of these intracellular parasites are able to survive and proliferate within haemocytes, the main cellular effectors of the immune system in molluscs. Two-dimensional electrophoresis was used to compare the haemolymph protein profile between Bonamia spp.-infected and non-infected oysters within 3 different stocks, a Galician stock of oysters selected for resistance against bonamiosis, a non-selected Galician stock and a selected Irish stock. Thirty-four proteins with a presumably relevant role in the oyster-Bonamia spp. interaction were identified; they were involved in major metabolic pathways, such as energy production, respiratory chain, oxidative stress, signal transduction, transcription, translation, protein degradation and cell defence. Furthermore, the haemolymph proteomic profiles of the non-infected oysters of the 2 Galician stocks were compared. As a result, 7 proteins representative of the non-infected Galician oysters selected for resistance against bonamiosis were identified; these 7 proteins could be considered as candidate markers of resistance to bonamiosis, which should be further assessed.


Assuntos
Haplosporídios/fisiologia , Hemolinfa/fisiologia , Ostrea/metabolismo , Ostrea/parasitologia , Animais , Regulação da Expressão Gênica , Hemócitos/metabolismo , Interações Hospedeiro-Parasita , Proteômica
9.
Genomics ; 110(6): 390-398, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29678683

RESUMO

European flat oyster (Ostrea edulis) production has suffered a severe decline due to bonamiosis. The responsible parasite enters in oyster haemocytes, causing an acute inflammatory response frequently leading to death. We used an immune-enriched oligo-microarray to understand the haemocyte response to Bonamia ostreae by comparing expression profiles between naïve (NS) and long-term affected (AS) populations along a time series (1 d, 30 d, 90 d). AS showed a much higher response just after challenge, which might be indicative of selection for resistance. No regulated genes were detected at 30 d in both populations while a notable reactivation was observed at 90 d, suggesting parasite latency during infection. Genes related to extracellular matrix and protease inhibitors, up-regulated in AS, and those related to histones, down-regulated in NS, might play an important role along the infection. Twenty-four candidate genes related to resistance should be further validated for selection programs aimed to control bonamiosis.


Assuntos
Haplosporídios , Hemócitos/metabolismo , Ostrea/genética , Infecções por Protozoários/genética , Transcriptoma , Animais , Regulação da Expressão Gênica , Hemócitos/imunologia , Ostrea/imunologia , Ostrea/metabolismo , Infecções por Protozoários/metabolismo
10.
J Fish Dis ; 41(1): 41-48, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28707705

RESUMO

The cockle Cerastoderma edule fishery has traditionally been the most important shellfish species in terms of biomass in Galicia (NW Spain). In the course of a survey of the histopathological conditions affecting this species in the Ria of Arousa, a haplosporidan parasite that had not been observed in Galicia was detected in one of the most productive cockle beds of Galicia. Uni- and binucleate cells and multinucleate plasmodia were observed in the connective tissue mainly in the digestive area, gills and gonad. The parasite showed low prevalence, and it was not associated with abnormal cockle mortality. Molecular identification showed that this parasite was closely related to the haplosporidan Minchinia mercenariae that had been reported infecting hard clams Mercenaria mercenaria from the Atlantic coast of the United States. The molecular characterization of its SSU rDNA region allowed obtaining a fragment of 1,796 bp showing 98% homology with M. mercenariae parasite. Phylogenetic analysis supported this identification as this parasite was clustered in the same clade as M. mercenariae from the United States and other M. mercenariae-like sequences from the UK, with bootstrap value of 99%. The occurrence of M. mercenariae-like parasites infecting molluscs outside the United States is confirmed.


Assuntos
Cardiidae/parasitologia , Doenças dos Peixes/parasitologia , Haplosporídios/classificação , Animais , DNA de Protozoário/análise , DNA Ribossômico/análise , Haplosporídios/genética , Filogenia , Infecções Protozoárias em Animais , Análise de Sequência de DNA , Espanha/epidemiologia
11.
Artigo em Inglês | MEDLINE | ID: mdl-29057216

RESUMO

Bonamiosis due to the parasite Bonamia ostreae has been associated with massive mortality outbreaks in European flat oyster stocks in Europe. As eradication and treatment are not possible, the control of the disease mainly relies on transfer restriction. Moreover, selection has been applied to produce resistant flat oyster families, which present better survival and lower prevalence than non-selected oysters. In order to better understand the mechanisms involved in resistance to bonamiosis, cellular and molecular responses of 2 oyster groups (selected oysters and wild-type oysters) were analyzed in the context of experimental injection and cohabitation infections. Cellular responses including non-specific esterases detection, ROS production and phagocytosis activity were analyzed by flow cytometry. Four genes homologous to those shown to be involved in immunity were selected (Inhibitor of apotosis OeIAP, Fas ligand OeFas-ligand, Oe-SOD, and OeEc-SOD) and monitored by quantitative reverse-transcription PCR (qRT-PCR). Infected oysters showed higher phagocytosis activity than controls. Infected selected oyster show a lower phagocytosis activity which might be a protection against the parasite infection. The expression of OeIAP and OeFas-ligand gene was significantly increased in selected oysters at 5 days post-injection. OeIAP gene expression appeared to be significantly increased in wild-type oysters at 8 days post-injection. Our results suggest that resistance to bonamiosis partly relies on the ability of the oysters to modulate apoptosis.


Assuntos
Resistência à Doença/genética , Haplosporídios/genética , Interações Hospedeiro-Parasita , Ostreidae/parasitologia , Infecções por Protozoários/parasitologia , Animais , Apoptose/genética , Expressão Gênica , Haplosporídios/isolamento & purificação , Hemócitos/metabolismo , Fagocitose/genética , Infecções por Protozoários/prevenção & controle , Espécies Reativas de Oxigênio/metabolismo , Fatores de Tempo
12.
J Vet Diagn Invest ; 29(5): 752-756, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28643557

RESUMO

Pooled testing of samples is a common laboratory practice to increase efficiency and reduce expenses. We investigated the efficacy of 2 published SYBR Green real-time PCR assays when used to detect the haplosporidian parasite Bonamia ostreae in pooled samples of infected oyster tissue. Each PCR targets a different gene within the B. ostreae genome: the actin 1 gene or the 18S rRNA gene. Tissue homogenates (150 mg) of the New Zealand flat oyster Ostrea chilensis were spiked with ~1.5 × 103 purified B. ostreae cells to create experimental pools of 3, 5, and 10. Ten positive replicates of each pool size were assayed twice with each PCR and at 2 different amounts of DNA template. The PCR targeting the actin 1 gene was unable to reproducibly detect B. ostreae in any pool size. Conversely, the 18S rRNA gene PCR could reproducibly detect B. ostreae in pools of up to 5. Using a general linear model, there was a significant difference in the number of pools that correctly detected B. ostreae between each PCR ( p < 0.01) and each pool size ( p < 0.01). It is likely that the single copy actin 1 gene is more likely to be diluted and not detected by pooling than the multi-copy 18S rRNA gene. Our study highlights that validation data are necessary for pooled sample testing because detection efficacy may not be comparable to individual sample testing.


Assuntos
Haplosporídios/isolamento & purificação , Haplosporídios/fisiologia , Ostrea/parasitologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Corantes Fluorescentes , Haplosporídios/genética , Interações Hospedeiro-Parasita , Nova Zelândia
13.
J Invertebr Pathol ; 148: 14-19, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28511901

RESUMO

Several stages of a haplosporidan parasite, including spores, were detected infecting three out of four specimens of the Pen Shell Pinna nobilis from the coast of Alicante (Western Mediterranean). A mortality event initiated few weeks before the sampling. The infection was systemic in the connective tissue, with free uni-nucleate stages and early plasmodia, whereas sporulation process took place in the digestive tubules disrupting them. Morphological details, by light and transmission electron microscopy, and PCR amplification confirmed that the parasite belongs to the haplosporidan group. Spores were pleomorphic, usually elongated ovoid, with round to elongated haplosporosomes-like in the sporoplasma. The operculum was situated in the apical zone of the wall, with an external lid, and the nucleus tended to be eccentric in the basal zone. Spore ornamentation was not observed. The single uninfected specimen appeared to be healthy. This is the first report of a haplosporidan parasite infecting a member of the Superfamily Pinnoidea and this is the first histopathological study of a mortality event in the endangered and protected P. nobilis.


Assuntos
Bivalves/parasitologia , Haplosporídios , Animais , Espanha
14.
Fish Shellfish Immunol ; 59: 331-344, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27815201

RESUMO

The flat oyster, Ostrea edulis, is one of the main farmed oysters, not only in Europe but also in the United States and Canada. Bonamiosis due to the parasite Bonamia ostreae has been associated with high mortality episodes in this species. This parasite is an intracellular protozoan that infects haemocytes, the main cells involved in oyster defence. Due to the economical and ecological importance of flat oyster, genomic data are badly needed for genetic improvement of the species, but they are still very scarce. The objective of this study is to develop a sequence database, OedulisDB, with new genomic and transcriptomic resources, providing new data and convenient tools to improve our knowledge of the oyster's immune mechanisms. Transcriptomic and genomic sequences were obtained using 454 pyrosequencing and compiled into an O. edulis database, OedulisDB, consisting of two sets of 10,318 and 7159 unique sequences that represent the oyster's genome (WG) and de novo haemocyte transcriptome (HT), respectively. The flat oyster transcriptome was obtained from two strains (naïve and tolerant) challenged with B. ostreae, and from their corresponding non-challenged controls. Approximately 78.5% of 5619 HT unique sequences were successfully annotated by Blast search using public databases. A total of 984 sequences were identified as being related to immune response and several key immune genes were identified for the first time in flat oyster. Additionally, transcriptome information was used to design and validate the first oligo-microarray in flat oyster enriched with immune sequences from haemocytes. Our transcriptomic and genomic sequencing and subsequent annotation have largely increased the scarce resources available for this economically important species and have enabled us to develop an OedulisDB database and accompanying tools for gene expression analysis. This study represents the first attempt to characterize in depth the O. edulis haemocyte transcriptome in response to B. ostreae through massively sequencing and has aided to improve our knowledge of the immune mechanisms of flat oyster. The validated oligo-microarray and the establishment of a reference transcriptome will be useful for large-scale gene expression studies in this species.


Assuntos
Bases de Dados Genéticas , Genoma , Haplosporídios/imunologia , Imunidade Inata , Análise de Sequência com Séries de Oligonucleotídeos , Ostrea/genética , Ostrea/parasitologia , Animais , Etiquetas de Sequências Expressas , Hemócitos/imunologia , Hemócitos/metabolismo , Hemócitos/parasitologia , Ostrea/imunologia , Análise de Sequência de DNA , Análise de Sequência de RNA , Transcriptoma
15.
Fish Shellfish Immunol ; 56: 322-329, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27431587

RESUMO

The in vitro model Ostrea edulis hemocyte - Bonamia ostreae is interesting to investigate host-parasite interactions at the cellular level. Indeed, this unicellular parasite infects the flat oyster Ostrea edulis and multiplies within hemocytes, the central effectors of oyster defenses. Apoptosis is a mechanism used by many organisms to eliminate infected cells. In order to study the potential involvement of this mechanism in the oyster response to B. ostreae, in vitro experiments were carried out by exposing hemocytes from the naturally susceptible oyster O. edulis and a resistant oyster species Crassostrea gigas to live and heat-inactivated parasites. Hemocyte apoptotic response was measured using a combination of flow cytometry and microscopy analyses. Whatever the host species was, the parasite was engulfed in hemocytes and induced an increase of apoptotic parameters including intracytoplasmic calcium concentration, mitochondrial membrane potential or phosphatidyl-serine externalization as well as ultrastructural modifications. However, the parasite appears more able to infect flat oyster than cupped oyster hemocytes and the apoptotic response was more important against live than dead parasites in the natural host than in C. gigas. Our results suggest that O. edulis specifically responds to B. ostreae by inducing apoptosis of hemocytes.


Assuntos
Apoptose , Haplosporídios/fisiologia , Interações Hospedeiro-Parasita , Ostrea/fisiologia , Ostrea/parasitologia , Animais , Citometria de Fluxo , Hemócitos/parasitologia , Hemócitos/fisiologia , Hemócitos/ultraestrutura , Microscopia Eletrônica de Transmissão
16.
Parasitology ; 143(9): 1119-32, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27263626

RESUMO

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.


Assuntos
Haplosporídios/classificação , Ostreidae/parasitologia , Platelmintos/classificação , Animais , Teorema de Bayes , Região do Caribe , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , Haplosporídios/genética , Haplosporídios/isolamento & purificação , Funções Verossimilhança , Ostreidae/classificação , Oceano Pacífico , Panamá , Zona do Canal do Panamá , Filogenia , Platelmintos/genética , Platelmintos/isolamento & purificação , Salinidade , Estações do Ano , Clima Tropical
17.
Dis Aquat Organ ; 118(1): 55-63, 2016 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-26865235

RESUMO

Previous reports of the haplosporidian parasite Bonamia ostreae have been restricted to the Northern Hemisphere, including Europe, and both eastern and western North America. This species is reported for the first time in New Zealand infecting the flat oyster Ostrea chilensis. Histological examination of 149 adult oysters identified 119 (79.9%) infected with Bonamia microcells. Bonamia generic PCR of several oysters followed by DNA sequencing of a 300 bp portion of the 18S rDNA gene produced a 100% match with that of B. ostreae. All DNA-sequenced products also produced a B. ostreae PCR-restriction fragment length polymorphism (PCR-RFLP) profile. Bonamia species-specific PCRs further detected single infections of B. exitiosa (2.7%), B. ostreae (40.3%), and concurrent infections (53.7%) with these 2 Bonamia species identifying overall a Bonamia prevalence of 96.6%. Detailed histological inspection revealed 2 microcell types. An infection identified by PCR as B. ostreae histologically presented small microcells (mean ± SE diameter = 1.28 ± 0.16 µm, range = 0.9-2 µm, n = 60) commonly with eccentric nuclei. A B. exitiosa infection exhibited larger microcells (mean ± SE diameter = 2.12 ± 0.27 µm, range = 1.5-4 µm, n = 60) with more concentric nuclei. Concurrent infections of both Bonamia species, as identified by PCR, exhibited both types of microcells. DNA barcoding of the B. ostreae-infected oyster host confirmed the identification as O. chilensis. A suite of other parasites that accompany O. chilensis are reported here for the first time in mixed infection with B. ostreae including apicomplexan X (76.5%), Microsporidium rapuae (0.7%) and Bucephalus longicornutus (30.2%).


Assuntos
Haplosporídios/fisiologia , Ostrea/parasitologia , Animais , Interações Hospedeiro-Parasita , Nova Zelândia
18.
J Invertebr Pathol ; 136: 7-9, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-26880159

RESUMO

In this study, the ability of oyster larvae, brooded in the pallial cavity of the parent oyster, to become infected in the pallial fluid, which is influenced by the brooding oyster and surrounding environment, was investigated. Larvae were collected over three summers from three areas around Ireland. Samples were screened for the presence of Bonamia ostreae DNA using PCR analysis. Four samples of larvae were found to be positive for B. ostreae DNA, though the parent oysters were negative for infection. Larvae may be able to acquire the pathogen from the water column during filter feeding or elimination of pseudo-faeces by the brooding adult.


Assuntos
Larva/microbiologia , Ostrea/microbiologia , Infecções por Protozoários/transmissão , Animais , Haplosporídios , Interações Hospedeiro-Parasita/fisiologia , Irlanda , Reação em Cadeia da Polimerase
20.
J Invertebr Pathol ; 130: 141-6, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26271576

RESUMO

In this study, we first report on the occurrence of Urosporidium sp., a haplosporidian hyperparasite infecting the trematode, Parvatrema duboisi, which parasitizes Manila clams, Ruditapes philippinarum on the west and south coasts of Korea. The larval P. duboisi infected by the sporocyst stage of Urosporidium sp. demonstrated numerous small yellowish spores in their tissues. The heavily infected metacercariae exhibited degenerate bodies and the larvae were often motionless. Clams heavily infected by the metacercariae of P. duboisi also displayed abnormal golden spots on the mantle tissue. In histology, different life stages of Urosporidium sp. could be identified, including the uni-nucleate, plasmodial, sporogonic stages and the acid fast mature spores released from the cyst. In scanning electron microscopy (SEM), the mature spore exhibited a semi-circular rim around the apical end and the orifice was covered internally with a flap. Loop-like filaments ornamentation was also identified from Urosporidium sp. in SEM, suggesting that Urosporidium sp. found in this study is a new member in the genus. Prevalence of Urosporidium sp.-infected trematodes in this study ranged from 2.5% to 24.0% in April 2010 and the infection was observed from 8 sampling sites out of the 26 sites surveyed on the west and south coasts.


Assuntos
Bivalves/parasitologia , Haplosporídios/parasitologia , Trematódeos , Animais , Microscopia Eletrônica de Varredura , Prevalência , República da Coreia
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