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1.
Bonamiosis and disseminated neoplasia (DN) are the most important diseases affecting cultured flat oysters (Ostrea edulis) in Galicia (NW Spain). Previous research of the response of O. edulis against bonamiosis by suppression subtractive hybridisation yielded a partial expressed sequence tag of tumour necrosis factor (TNF) and allograft inflammatory factor (AIF), as well as the whole open reading frame for dermatopontin and vesicle-associated membrane (VAMP). Herein, the complete open reading frames of TNF and AIF genes were determined by the rapid amplification of cDNA, and the deduced amino acid sequences of the four genes were characterised. Phylogenetic relationships for each gene were studied using maximum likelihood parameters. Quantitative-PCR assays were also performed in order to analyse the modulation of the expression of these genes by bonamiosis and disseminated neoplasia. Gene expression profiles were studied in haemolymph cells and in various organs (gill, gonad, mantle and digestive gland) of oysters affected by bonamiosis, DN, and both diseases with regard to non-affected oysters (control). TNF expression in haemolymph cells was up-regulated at heavy stage of bonamiosis but its expression was not affected by DN. AIF expression was up-regulated at heavy stage of bonamiosis in haemolymph cells and mantle, which is associated with heavy inflammatory response, and in haemolymph cells of oysters affected by DN. AIF expression was, however, down-regulated in other organs as gills and gonads. Dermatopontin expression was down-regulated in haemolymph cells and digestive gland of oysters affected by bonamiosis, but DN had no significant effect on its expression. Gills and gonads showed up-regulation of dermatopontin expression associated with bonamiosis. There were significant differences in the expression of TNF and VAMP depending on the bonamiosis intensity stage whereas no significant differences were detected between light and heavy severity degrees of DN for the studied genes. VAMP expression showed also differences among haemolymph cells and the organs studied. The occurrence of both diseases in oysters involved haemolymph cell gene expression patterns different from those associated to each disease separately: no significant effect was observed in TNF expression, dermatopontin was up-regulated and marked up-regulation of AIF and VAMP was recorded, which suggests a multiplier effect of the combination of both diseases for the latter two genes.  相似文献   

2.
A research project to compare productive traits (growth and mortality), disease susceptibility and immune capability between Ostrea edulis stocks was performed. This article reports the results on the immune capability and its relation with infection by the intrahaemocytic protozoan Bonamia ostreae. Four to five oyster spat families were produced from each of four European flat oyster populations (one from Ireland, one from Greece and two from Galicia, Spain) in a hatchery. The spat were transferred to a raft in the Ría de Arousa (Galicia) for on growing for 2 years. Total haemocyte count (THC) and differential haemocyte count (DHC) were estimated monthly through the second year of growing-out. Three types of haemocytes were distinguished: granulocytes (GH), large hyalinocytes (LHH) and small hyalinocytes (SHH). Significant correlations between the mean relative abundance of GH and SHH of the families and the mean prevalence of B. ostreae, the overall incidence of pathological conditions and the cumulative mortality of the families were found; these correlations supported the hypothesis that high %GH and low %SHH would enhance oyster immune ability and, consequently, would contribute to lower susceptibility to disease and longer lifespan. Infection by B. ostreae involved a significant increase of circulating haemocytes, which affected more markedly the LHH type. The higher the infection intensity the higher the %LHH. This illustrates the ability of B. ostreae to modulate the immune responses of the O. edulis to favour its own multiplication. A significant reduction of the phenoloxidase activity in the haemolymph of oysters O. edulis infected by B. ostreae was observed. Nineteen enzymatic activities in the haemolymph of O. edulis and Crassostrea gigas (used as a B. ostreae resistant reference) were measured using the kit api ZYM®, Biomerieux. Qualitative and quantitative differences in enzyme activities in both haemocyte and plasma fractions between B. ostreae noninfected O. edulis from different origins were recorded. However, no clear positive association between enzyme activity and susceptibility to bonamiosis was found. The only enzyme detected in the resistant species C. gigas that was not found in the susceptible one O. edulis was β-glucosidase (in plasma). B. ostreae infected O. edulis showed significant increase of some enzyme activities and the occurrence of enzymes that were not detected in noninfected oysters. These changes could be due to infection-induced enzyme synthesis by the host or to enzyme synthesis by the parasite.  相似文献   

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5.
Range expansion and population establishment of individual species can have significant impacts on previously established food webs and predator-prey dynamics. The stone crab (Menippe spp.) is found throughout southwestern North Atlantic waters, from North Carolina through the Gulf of Mexico and the Central American Caribbean, including the Greater Antilles. Recent observations suggest that stone crabs have become better established on certain oyster reefs in North Carolina than in the early 1900s when they we first observed in NC. To assess the predatory impact of stone crabs on oysters, we (1) quantified stone crab densities on subtidal oyster reefs in Pamlico Sound, NC using scuba surveys, and (2) conducted laboratory predation experiments to assess the functional response of stone crabs to varying densities of oysters. We then (3) analyzed previously unpublished functional response data on another important oyster predator, the mud crab Panopeus herbstii. Finally, we (4) compared and contrasted potential predatory impacts of stone, mud and blue crabs (Callinectes sapidus). The functional response data and analyses for both stone crabs and mud crabs were consistent with a type II functional response. Mud crabs, on a m2 basis, inflicted the highest proportional mortality on oysters over a 24 hour period, followed by stone and then blue crabs. Proportional mortality did not vary significantly with oyster size; however, relatively small and large oysters were consumed disproportionately less than medium-sized oysters, likely due to the mechanical inability of stone crabs to handle small oysters, and the inability to crush large oysters. Although stone crabs appear to be established in Pamlico Sound at densities equivalent to densities in other systems such as the U.S. Florida Panhandle, their predatory activities on oysters are not expected to have as significant a negative impact on oyster populations compared to other resident predators such as mud crabs.  相似文献   

6.
The oyster ovarian parasite Marteilioides chungmuensis has been reported from Korea and Japan, damaging the oyster industries. Recently, Marteilioides-like organisms have been identified in other commercially important marine bivalves. In this study, we surveyed Marteilioides infection in the Manila clam Ruditapes philippinarum, Suminoe oyster Crassostrea ariakensis, and Pacific oyster Crassostrea gigas, using histology and Marteilioides-specific small subunit (SSU) rDNA PCR. The SSU rDNA sequence of M. chungmuensis (1716 bp) isolated from C. gigas in Tongyoung bay was 99.9% similar to that of M. chungmuensis reported in Japan. Inclusions of multi-nucleated bodies in the oocytes, typical of Marteilioides infection, were identified for the first time in Suminoe oysters. The SSU rDNA sequence of a Marteilioides-like organism isolated from Suminoe oysters was 99.9% similar to that of M. chungmuensis. Marteilioides sp. was also observed from 7 Manila clams of 1840 individuals examined, and the DNA sequences of which were 98.2% similar to the known sequence of M. chungmuensis. Unlike Marteilioides infection of Pacific oysters, no remarkable pathological symptoms, such as large multiple lumps on the mantle, were observed in infected Suminoe oysters or Manila clams. Distribution of the infected Manila clams, Suminoe oysters and Pacific oysters was limited to small bays on the south coast, suggesting that the southern coast is the enzootic area of Marteilioides infection.  相似文献   

7.
Several Vibrio species are known to be pathogenic to the Pacific oyster Crassostrea gigas. Survival varies according to pathogen exposure and high mortality events usually occur in summer during gametogenesis. In order to study the effects of gametogenetic status and ploidy (a factor known to affect reproduction allocation in oysters) on vibriosis survival, we conducted two successive experiments. Our results demonstrate that a common bath challenge with pathogenic Vibriosplendidus and Vibrio aestuarianus on a mixture of mature, spawning and non-mature oysters can lead to significant mortality. Previous bath challenges, which were done using only non-mature oysters, had not produced mortality. Immunohistochemical analyses showed the affinity of Vibrio for gonadic tissues, highlighting the importance of sexual maturity for vibriosis infection processes in oysters. Mortality rate results showed poor repeatability between tanks, however, in this bath challenge. We then tested a standardized and repeatable injection protocol using two different doses of the same combination of two Vibrio species on related diploid and triploid oysters at four different times over a year. Statistical analyses of mortality kinetics over a 6-day period after injection revealed that active gametogenesis periods correspond to higher susceptibility to vibriosis and that there is a significant interaction of this seasonal effect with ploidy. However, no significant advantage of triploidy was observed. Triploid oysters even showed lower survival than diploid counterparts in winter. Results are discussed in relation to differing energy allocation patterns between diploid and triploid Pacific oysters.  相似文献   

8.
Disseminated neoplasia (DN) has been detected in cockles from various beds in Galicia (NW Spain). A study was performed to characterise cockle neoplastic cell ultrastructure and to evaluate the effect of this disease at different severity stages on various haemolymph cell parameters. Examination of cockle neoplastic cells with transmission electron microscopy (TEM) showed round shapes and a lack of pseudopods, a high nucleus:cytoplasm diameter ratio, Golgi complexes, abundant mitochondria, ribosomes, and numerous endoplasmic reticulum tubes and electron-lucent vesicles. Various haemolymph cell parameters (cell mortality, non-specific esterase and lysosome biovolume, reactive oxygen intermediates [ROI] production, phagocytosis ability, intracellular Ca2+ and actin levels) were compared between DN severity categories by flow cytometry; haemocyte mortality, non-specific esterase activities and lysosome biovolume were found to be higher with increasing DN severity. The phagocytic ability of neoplastic cells was sharply reduced with regard to haemocytes. The cytoplasmic-free Ca2+ level was higher and actin content lower in haemolymph cells of diseased cockles compared to unaffected ones. A significant increase in ROI production was detected in later stages of disease progression.  相似文献   

9.
An epidemiological survey was performed to know the infection status of oysters with Gymnophalloides seoi metacercariae in 7 islands of the West Sea known as the habitat of paleartic oystercatchers, Haematopus ostralegus osculans, in Korea. The surveyed areas were Aphaedo (Shinan-gun, Chollanam-do), Jangjado, Sonyudo and Munyodo (Okdo-myon, Kunsan-shi, Chollabuk-do), Yubudo (Changhang-up, Sochon-gun Chungchongnam-do), and Polumdo and Chumoondo (Sodo-myon, Kangwha-gun. Inchon-shi). The oysters collected from Aphaedo, the known endemic focus, were examined monthly from August 1995 to October 1996 for observation of any seasonal variation of the metacercarial density. The average metacercarial burden was 761-2,077 by month, but the seasonal variation of the metacercarial density was not obvious. A total of 54 metacercariae was detected in 63 oysters collected from Yubudo. Out of 30 oysters from Sonyudo, 25 (83.3%) were infected with 1-66 metacercariae (12.6 in average). All of 50 oysters (100%) from Munyodo were infected with 3-162 metacercariae (53.5 in average). Only 4 metacercariae were detected in 100 oysters from Chumoondo. However, no metacercariae were found in 55 oysters from Jangjado and 50 oysters from Polumdo. From the above results, it was confirmed that G. seoi is still highly prevalent in oysters from Aphaedo, and several islands of the West Sea known as the habitats of paleartic oystercatchers are new endemic areas of this fluke.  相似文献   

10.
A comparison among various histological techniques for the detection of the parasite Bonamia ostreae in oysters Ostrea edulis was performed to evaluate their sensitivity and suitability for different purposes. The comparison involved examination of histological sections, tissue imprints from gills, digestive gland, gonad and heart, and haemolymph cell monolayers, prepared through various protocols. Every technique produced some false negative. The haemolymph cell monolayers were more sensitive than tissue imprints and histological sections. Heart imprints provided the highest sensitivity among tissue imprints. Examination of histological sections was among the least sensitive techniques. Four procedures for estimation of infection intensity were compared. Some differences in accuracy for the estimation of infection intensity between haemolymph cell monolayers and histological sections (HS) were detected: there was a very good agreement when the infection appeared low or heavy in HS but it was not so good in the remaining cases. The results suggest the need for a critical review of the recommendations of the "Office Internationale des Epizooties" and the European Union for diagnosis of bonamiosis.  相似文献   

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The aim of this study was to assess the effect of dietary eicosapentaenoic acid (20:5n-3) on hemocyte parameters such as hemocyte concentration, phagocytosis, and non-stimulated reactive oxygen species (ROS) production in Pacific oysters Crassostrea gigas, as well on proximate biochemical and fatty acid compositions. One-year-old oysters (C. gigas) were fed T-Isochrysis aff. galbana (T-Iso), which is low in 20:5n-3, either alone or with supplements of a lipid emulsion rich in 20:5n-3 at 1%, 10% or 50% (dry weight of the algal ration) for up to 7 weeks. Changes in gill fatty acid composition demonstrated that the lipid emulsion was well ingested by oysters during the dietary conditioning. Biochemical analysis indicated that oysters fed supplements of 50% and, to a lesser extent, 10% lipid emulsions had a higher total lipid content compared with oysters fed other diets, suggesting a more advanced reproductive status for the oysters fed high doses of lipid emulsion. Moreover, some oysters in these two treatment groups spawned during the last three weeks of the seven-week feeding experiment. Lipid supplements had a significant influence on hemocyte concentration, phagocytic index and non-stimulated hemocyte ROS production. After 4 weeks, highest hemocyte concentrations were found in oysters fed on a supplement of 50% lipid emulsion compared with those fed on other diets but the hemocytes derived from these oysters had the lowest short-term phagocytic index. After 7 weeks of dietary conditioning, the ROS production in non-stimulated hemocytes of oysters fed 10% and 50% lipid emulsion declined. These results suggested that 20:5n-3, and perhaps its eicosanoid metabolites, affected oyster hemocyte functions; however, the reproductive status of oysters may also have interfered with the 20:5n-3 dietary effect.  相似文献   

13.
QX disease is a fatal disease in Sydney rock oysters caused by the protozoan parasite Marteilia sydneyi. The current study investigates the phagocytosis of M. sydneyi by Sydney rock oyster hemocytes. It also compares the in vitro phagocytic activities of hemocytes from oysters bred for QX disease resistance (QXR) with those of wild-type oysters. After ingestion of M. sydneyi, hemocyte granules fused with phagosome membranes and the pH of phagosomes decreased. Significantly (p = <0.05) more phagosomes in QXR hemocytes showed obvious changes in pH within 40 min of phagocytosis, when compared with wild-type hemocytes. Phenoloxidase deposition was also evident in phagosomes after in vitro phagocytosis. Most importantly, ingested and melanised M. sydneyi were detected in vivo among hemocytes from infected oysters. Overall, the data suggest that Sydney rock oyster hemocytes can recognise and phagocytose M. sydneyi, and that resistance against QX disease may be associated with enhanced phagolysosomal activity in QXR oysters.  相似文献   

14.
We have identified quantitative trait loci (QTL) in the flat oyster (Ostrea edulis) for resistance to Bonamia ostreae, a parasite responsible for the dramatic reduction in the aquaculture of this species. An F2 family from a cross between a wild oyster and an individual from a family selected for resistance to bonamiosis was cultured with wild oysters injected with the parasite, leading to 20% cumulative mortality. Selective genotyping of 92 out of a total of 550 F2 progeny (i.e., 46 heavily infected oysters that died and 46 parasite-free oysters that survived) was performed using 20 microsatellites and 34 amplification fragment length polymorphism primer pairs. Both a two-stage testing strategy and QTL interval mapping methods were used. The two-stage detection strategy had a high power with a low rate of false positives and identified nine and six probable markers linked to genes of resistance and susceptibility, respectively. Parent-specific genetic linkage maps were built for the family, spanning ten linkage groups (n = 10) with an observed genome coverage of 69–84%. Three QTL were identified by interval mapping in the first parental map and two in the second. Good concordance was observed between the results obtained after the two-stage testing strategy and QTL mapping.  相似文献   

15.
Oysters Crassostrea gigas were placed at water supply canals of three shrimp farms in Guasave, Mexico where WSSV outbreaks occur. Animals were sampled through April-August and September-December to detect WSSV DNA.By using three different PCR protocols, only oysters from a farm undergoing a WSSV outbreak were found WSSV-positive in gills and digestive gland. Two WSSV amplicons were sequenced and they corresponded over 99% to WSSV genome segments. Results showed that oysters can capture WSSV particles suspended in water. Susceptibility of oysters to WSSV infection and their role as a carrier remain to be determined.  相似文献   

16.
Samples from the field and laboratory exposure to Mikrocytos mackini (a tiny protistan parasite of unknown taxonomic affiliation) confirmed that juvenile Pacific oysters (Crassostrea gigas) are susceptible to infection and the resulting disease. In the laboratory bath exposure experiment, a prevalence of infection approaching 100% and mortalities were observed in the small oysters (about 18 mm in shell length). However, in the same laboratory exposure experiment, similar aged geoduck clams (Panope abrupta, about 8mm in shell length) were resistant to infection. The main route of infection in the oysters appeared to be via the digestive tract and possibly the gills where the parasite multiplied within host cells. Other tissues such as the adductor muscle and vesicular connective tissue were subsequently colonized. Although the infection resulted in the mortality of some oysters, others appeared to overcome the disease.  相似文献   

17.
Pacific oysters Crassostrea gigas (Thunberg, 1793) were introduced to the northern Wadden Sea (North Sea, Germany) by aquaculture in 1986 and finally became established. Even though at first recruitment success was rare, three consecutive warm summers led to a massive increase in oyster abundances and to the overgrowth of native mussel beds (Mytilus edulis L.). These mussels constitute biogenic reefs on the sand and mud flats in this area. Survival and growth of the invading C. gigas were investigated and compared with the native mussels in order to predict the further development of the oyster population and the scope for coexistence of both species. Field experiments revealed high survival of juvenile C. gigas (approximately 70%) during the first three months after settlement. Survival during the first winter varied between > 90% during a mild and 25% during a cold winter and was independent of substrate (i.e., mussels or oysters) and tide level. Within their first year C. gigas reached a mean length of 35-53 mm, and within two years they grew to 68-82 mm, which is about twice the size native mussels would attain during that time. Growth of juvenile oysters was not affected by substrate (i.e., sand, mussels, and other oysters), barnacle epibionts and tide level, but was facilitated by fucoid algae. By contrast, growth of juvenile mussels was significantly higher on sand flats than on mussel or oyster beds and higher in the subtidal compared to intertidal locations. Cover with fucoid algae increased mussel growth but decreased their condition expressed as dry flesh weight versus shell weight. High survival and growth rates may compensate for years with low recruitment, and may therefore allow a fast population increase. This may lead to restrictions on habitat use by native mussels in the Wadden Sea.  相似文献   

18.
The feeding and biodeposition rates of small (2.4 cm) and large (5.2 cm) Crassostrea virginica were differently affected in experiments using flowing systems by moderate (292 μ C/1 sea water) and high (487 μ C/1 sea water) concentrations of the following phytoplankton: the diatoms, Thalassiosira pseudonana (Hasle & Heindal) (Cyclotella nana), Skeletonema costatum (Greville) Cleve, Nitzschia closterium (Ehrenberg) Wm. Smith, and the green alga, Dunaliella tertiolecta Butcher.The average feeding rate of the oysters on the different phytoplankton was: Dunaliella < Nitzschia < Skeletonema < Thalassiosira. In contrast, oysters fed on Thalassiosira had the lowest rate of biodeposition (1.626 × 10?3 g ash-free dry wt/g dry flesh wt per day) but oysters cultured on Nitzschia showed the highest rate of biodeposition (2.952 × 10?3 units).The response of the feeding rate of the two sizes of oysters to food concentration differed according to the algal food source. Oysters fed on Thalassiosira and Skeletonema exhibited a higher feeding rate at a higher food level: in contrast, oysters fed on Nitzschia showed no significant differences and those cultured on Dunaliella a depressed feeding rate at the higher food concentration. We observed no differences in feeding rate between small and large oysters fed on Skeletonema and Dunaliella, but small oysters had a higher feeding rate than large oysters when cultured on Thalassiosira and a lower feeding rate when fed on Niitzschia. The smaller size of Thalassiosira and larger size of Nitzschia might account for these differences in the feeding efficiency of small oysters.The biodeposition rates of both sizes of oysters were also affected by food concentration of the different algae. At the high food concentration with Skeletonema, Nitzschia, and Dunaliella, all oysters showed a marked increase in the rate of biodeposition, but oysters fed on Thalassiosira exhibited no great change. On all species of phytoplankton, the larger oysters produced greater biodeposition than the smaller oysters.These differences in response of feeding and biodeposition rates are important to our knowledge of food chain dynamics and may be used to optimize aquaculture systems.  相似文献   

19.
Within the framework of a national scientific program named “MORtalités ESTivales de l'huître creuse Crassostrea gigas” (MOREST), a family-based experiment was developed to study the genetic basis of resistance to summer mortality in the Pacific oyster, Crassostrea gigas. As part of the MOREST project, the second generation of three resistant families and two susceptible families were chosen and pooled into two respective groups: “R” and “S”. These two groups of oysters were conditioned for 6 months on two food levels (4% and 12% of oyster soft-tissue dry weight in algal dry weight per day) with a temperature gradient that mimicked the Marennes-Oléron natural cycle during the oyster reproductive period. Oyster mortality remained low for the first two months, but then rapidly increased in July when seawater temperature reached 19 °C and above. Mortality was higher in “S” oysters than in “R” oysters, and also higher in oysters fed the 12% diet than those fed 4%, resulting in a decreasing, relative order in cumulative mortality as follows; 12% “S” > 12% “R” > 4% “S” > 4% “R”. Although the observed mortality rates were lower than those previously observed in the field, the mortality differential between “R” and “S” oysters was similar. Gonadal development, estimated by tissue lipid content, followed a relative order yielding a direct, positive relationship between reproductive effort and mortality as we reported precedently by quantitative histology. Regarding hemocyte parameters, one of the most striking observations was that reactive oxygen species (ROS) production was significantly higher in “S” oysters than in “R” oysters in May and June, regardless of food level. The absence of known environmental stress under these experimental conditions suggests that the ROS increase in “S” oyster could be related to their higher reproductive activity. Finally, a higher increase in hyalinocyte counts was observed for”S” oysters, compared to “R” oysters, in July, just before mortality. Taken together, our results suggest an association of genetically based resistance to summer mortality, reproductive strategy and hemocyte parameters.  相似文献   

20.
The effects of season and qualitative selection capacity on trophic relationships between two sympatric invasive suspension-feeders, Crepidula fornicata and Crassostrea gigas, were investigated in Bourgneuf Bay (France) from January 2003 to June 2004. Carbon and nitrogen stable isotopic deviations, δ13C and δ15N, of common Atlantic slippersnails and Pacific oysters were analysed relative to isotopic composition and availability of end-members.Slippersnail deviations were less variable over the sampling period compared with those of oysters. Significant differences between δ13C and δ15N of C. fornicata and C. gigas were found from winter to early summer, and linked to major isotopic changes in oysters. We identified three distinct seasonal periods: January to March when oysters were 15N-enriched compared to slippersnails and to themselves at other times of the year, April to June-July when oysters showed a 15N-depletion and a more marked 13C-depletion compared to slippersnails and to themselves at other times of the year, and July-August to December when both species presented similar carbon and nitrogen deviations. Species-specific differences in qualitative selection capability may explain these seasonal differences in isotopic deviations. Whereas the isotopic composition of the indiscriminate suspension-feeding slippersnails reflects the composition of the seston throughout the year, the oyster is capable of qualitative selection. The oyster isotopic compositions are consistent with a facultative activation of selection mechanisms under conditions of qualitative and quantitative food limitation, notably the preferential ingestion and assimilation of the dominant organic source in the suspended pool.We conclude that C. fornicata and C. gigas are trophic competitors only in winter and spring at this site, where detrital end-members are major POM components. These results underscore (1) the importance of long-term (annual) studies in the evaluation of potential trophic competition, and (2) the necessity to include the qualitative selection capacities of suspension-feeders in future interpretations of trophic relationships in marine coastal ecosystems.  相似文献   

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