Reproduction, condition and mortality of the Pacific oyster Crassostrea gigas (Thunberg) in Sonora, México

Starting in 1997, mortality outbreaks in cultured oysters Crassostrea gigas have been reported in northwestern México. Previous studies have shown that massive die‐offs result from multi‐factor processes related to reproduction of the species. We studied the reproductive cycle and the condition index of cultured oysters in the coastal lagoon of El Soldado, Sonora, as well as the relationship of the life cycle with environmental parameters. We used oocyte diameter to determine reproductive stages and the condition index to describe the physiological state of oysters. Additionally, the temperature, salinity, seston and chlorophyll a were recorded at the study site. The results showed that C. gigas began accelerated reproductive activity in March under the influence of high temperature and increased concentrations of food. No spawning events were recorded and gametes were reabsorbed within the gonad in September and October. The results showed a period of nutrient storage during autumn–winter and another period of gamete production in spring–summer. A mortality event occurred at the end of winter, tied to significant increases in temperature and availability of food and in accelerated reproductive activity and high condition index. These conditions were very similar to those reported in other countries during summer die‐offs of C. gigas.     

Quantitative measurement of reproductive output in the American oyster, Crassostrea virginica (Gmelin), using an enzyme-linked immunosorbent assay (ELISA)

Abstract. A quantitative gonadal index was developed for oysters, Crassostrea virginica (Gmelin. 1791), using polyclonal antibodies from eggs and sperm. Percoll used in the purification of oyster eggs and sperm greatly improved the purity of antigens compared to filtering the egg or sperm through a fine mesh only. The antigen-antibody reaction was tested with indirect sandwich ELISA using alkaline phosphatase-conjugated goat anti-rabbit igG as a secondary antibody. Rabbit anti-oyster egg IgG and anti-oyster sperm IgG initially exhibited a weak cross-reactivity over somatic tissue. Absoring with acetone-dried oyster tissue powder removed this cross-reactivity. Both antisera exhibited strong specific immunological reactions to oyster eggs or sperm respectively. The quantity of eggs or sperm was measured using ELISA and a quantitative gonadosomatic index (dry wt of egg or sperm/dry wt oyster) (GSI) calculated. GSI from ELISA correlated with gonadal stage measured histologically. Monthly mean GSI of female oysters was highest during late spring to early summer (0·157–0·201) and lowest during early winter to early spring (0·002-0·000). Maximum GSI observed during the study was 0·422 for female oysters and 0·446 for male oysters. Female oysters produce 3·7–65·4 million eggs, with an average of 21·1 million during each spawning. A positive correlation was observed between the number of eggs produced and oyster size; the number of eggs in the gonad increased as oyster size (i.e. total dry wt) increased (r= 0·67); however, the relationship was non-linear. Large oysters contained proportionally fewer eggs. Prevalence of Perkinsus marinus parasitism was high, 90–100%, during the study, as was weighted incidence, 1·33 to 2·67, No statistically significant correlation was observed between infection intensity and the per cent weight of oyster eggs or egg number.     

Effects of tributyltin and hypoxia on the progression of Perkinsus marinus infections and host defence mechanisms in oyster, Crassostrea virginica (Gmelin)

Oysters, Crassostrea virginica (Gmelin), naturally infected by the protozoan parasite Perkinsus marinus in the field were exposed for 6 weeks to tributyltin (TBT), hypoxia, or to both stressors simultaneously. The TBT-exposed oysters continuously bioaccumulated TBT, reaching about 4 mg kg^??1 dry weight by 6 weeks; hypoxic oysters were exposed to water containing an average dissolved oxygen level of about 3 mg L^??1. Untreated control oysters suffered about 30% cumulative mortality by 6 weeks as a result of the progression of their P. marinus infections. The TBT treatment alone produced no additional mortality; however, cumulative mortality in hypoxic oysters was elevated. Mortality among oysters receiving both TBT and hypoxia significantly exceeded that caused by either stressor alone, suggesting a synergistic effect. In an attempt to identify immunotoxicological mechanisms underlying stress-related augmentation of P. marinus infections, defence-related immune functions were measured at 3 and 6 weeks in control and treated oysters. In general, the total number of haemocytes increased as the infections progressed, and the TBT and hypoxic treatments also caused significant additional increments in some samples. However, oxygen-dependent (reactive oxygen species) and oxygen-independent (lysozyme) antimicrobial host defence mechanisms appeared to be largely unaffected by TBT and/or hypoxia. This may be explained by the death of those oysters with marked immunological lesions prior to sampling or by the actual lack of treatment effects.     

Quantitative quality evaluation of Eastern oyster (Crassostrea virginica) cultured by two different methods

In order to evaluate the quality of oysters and whether a production method affects quality or not, a set of objective quantitative quality measures was developed. Eastern oysters (Crassostrea virginica) were cultured using two different treatments (never desiccated or desiccated daily with tidal exposure) and the meats were tested by textural, physico‐chemical and chemical analyses over a 25‐day cold storage period. Texture analyses parameters (cutting force and chewiness on oyster adductor muscles) were strongly correlated with storage time before the death of oysters and could be used as a quality indicator for oyster. Total volatile basic nitrogen (TVBN) could be used as freshness indicator for raw Eastern oysters, with the acceptability of 11 mg/100 g. Models for evaluating the quality of oysters were established that reflect the impact of a biofouling treatment on oyster's shelf life and texture attributes. The methods and quality indicators developed in this study were effective in evaluating the quality and freshness of Eastern oysters objectively, and could serve as routine quality check of oyster meat for the industry.     

Hot water immersion lowers survival,shell growth rate and lysosomal membrane stability of oysters Crassostrea virginica (Gmelin)

This study compares the effect of two anti‐fouling treatments, hot water immersion (15 s at 60°C) and air drying (72 h) on the physiological status of the Eastern oyster Crassostrea virginica. The negative impact of hot water immersion was greater than that of air drying, but varied depending on the initial size of the oysters (40 vs. 60 mm shell height) and the time of the year (June vs. August). Groups treated with hot water exhibited a higher proportion of haemocytes with destabilized lysosomal membranes (HDLM; 47.5 ± 3.1%) than those exposed to air drying (37.5 ± 2.9%). This suggests that the oyster immunocompetency may be lowered by hot water immersion. Overall, the large oysters had lower HDLM values (32.9 ± 3.5%) than the small individuals in June (45.7 ± 2.8%) but similar values in August (46.6 ± 3.5%). Small oysters subjected to hot water immersion in June exhibited a 50% reduction in shell growth and a 50% mortality rate after one month. Our results indicate that air drying is more suitable than hot water immersion as an anti‐fouling treatment for <45 mm oysters.     

A comparison of growth and survival of subtidal Crassostrea virginica (Gmelin) in South Carolina salt marsh impoundments

Four South Carolina salt marsh impoundments and their associated tidal creeks were assessed for the culture of subtidal Crassostrea virginica. The impoundments were chosen primarily for their diversity and ranged from old large impoundments with appreciable tidal exchange and surrounded by extensive low marsh, to new small impoundments with little tidal exchange and surrounded by maritime forest. Floating and bottom hardware cloth trays (1.22 × 0.61 × 0.14 m) each holding 200 seed oysters (initial length, $\text{y}\text{= 43.8}\text{mm}$) were placed at each location and sampled monthly for growth and survival. Coincidental monthly estimates of primary production (¹⁴C), phytoplankton concentrations and total organic carbon were performed. Ancillary data collected biweekly at all locations included standard hydrographic information (temperature, salinity, pH and turbidity) and nutrient determinations (nitrates, nitrites, orthophosphates and silicates).Results indicated that over a 6-month period (October–April) growth at all locations was significantly (α = 0.001) greater in ponds than in adjacent creeks and greater in floating than bottom trays. Growth means ranged from as little as 1.00 mm/month in the Wando River, to median values of approximately 2.25 mm/month in tidal creeks, to a relatively high growth rate of 3.11 mm/month in Blue Heron Pond (Kiawah Island). Survival was high in all areas ranging from 85.0% at Blue Heron Pond to 94.5% at Kiawah Creek. There was no significant difference in survival in comparisons among locations. A direct correlation of growth in oysters, primary production and phytoplankton biomass was established. This relationship was reiterated by indications of an inverse correlation between nutrient concentrations and growth.     

Stress response of Crassostrea virginica (Gmelin, 1791) oysters following a reciprocal transfer between upriver and downriver sites

This study was designed to assess the physiological impact of transferring oysters between upriver and downriver aquaculture sites, a common practice in North America that is primarily aimed at reducing disease infections and predation on cultured stocks. In May 2009, oysters (Crassostrea virginica) were reciprocally transferred between an upriver and a downriver site in the Richibucto estuary in eastern Canada. Mortality, tissue and cellular stress responses were subsequently evaluated in August and October 2009. Overall, oyster mortality remained low (~5%) throughout the 5‐month study period with no significant difference between sites or oyster sources. However, by October oysters reared at the upriver site, regardless of their origin, had significantly higher levels of lysosomal membrane destabilization (63.6%, SE = 1.9) and digestive tubule atrophy (33.3–42.4%, SE = 3.6) than oysters reared at the downriver site (47.5%, SE = 1.8; 15.6–19.1%, SE = 3.8 respectively). They also exhibited a greater salinity differential between their mantle/haemolymph fluids and the ambient seawater, possibly indicating more restricted exchange with the environment. In general, the transfer of upriver oysters to a downriver site had a positive impact, i.e. lower levels of lysosomal destabilization and tubule atrophy, whereas transfer of downriver oysters upriver had the opposite effect. These results suggest that upriver environmental conditions negatively impact cellular and tissue integrity in oysters without leading to mortality during the summer–autumn period.     

Growth,morphometrics and nutrient content of farmed eastern oysters,Crassostrea virginica (Gmelin), in New Hampshire,USA

When harvested, oysters represent a removal from the ecosystem of nutrients such as nitrogen (N) and carbon (C). A number of factors potentially affect nutrient content, but a quantitative understanding across the geographical range of the eastern oysters is lacking. This study was designed to quantify the relationships among various metrics of farmed eastern oysters near its northern geographical range focusing on nutrient content. Hatchery‐reared oysters were deployed in polyethylene bags at six sites, and were measured on multiple occasions from 2010 to 2012. A quadratic polynomial fit to the combined datasets for shell height indicated that on average a ‘cocktail’ size oyster (63 mm shell height) would be reached after 2 year, and ‘regular’ size (76 mm) would require 3 year. There were significant differences in growth rates and oyster nutrient content among the sites; means for %N in soft tissue ranged from 6.9 to 8.6, and 0.07 to 0.18 in shell. Per cent N in soft tissue and shell were highest at two sites at the mouths of rivers with elevated dissolved inorganic N concentrations in the water. Grand means (all sites, seasons and years combined) of soft tissue N and C for regular size oysters were 7.3% and 38.5%, respectively; and for shell N and C were 0.13% and 12.0% respectively. Our study extends the range of data on nutrient content of the eastern oyster to northern New England, and indicates that oyster size, seasonality, and nutrient concentration in ambient water potentially affect %N and %C content of oysters.     

Resistance of Dermo in eastern oysters, Crassostrea virginica (Gmelin), of North Carolina but not Chesapeake Bay Heritage

Growth, intensity of Perkinsus marinus (Levine) infection, and survival of synchronously spawned North Carolina (NC) and Chesapeake Bay‐heritage (CB) oysters, Crassostrea virginica (Gmelin) were evaluated under standard tray culture conditions at several sites in both regions (Wye River, Maryland; Mobjack Bay, Virginia; Pamlico River, NC and Bogue Banks, NC). Infection prevalence reached 100% in oysters held at all high‐ and moderate‐salinity sites, at which time the CB strain ceased to grow. Shortly after growth ceased, CB oysters exhibited mortality that rapidly progressed to 100%. Unlike the CB strain, growth continued in the NC strain despite high P. marinus prevalence. When mortality did occur in the NC strain, at a reduced rate of 37–40%, it was associated with higher intensity of P. marinus than the infection intensity correlated with death of CB oysters. At the low‐salinity site in NC, P. marinus infection persisted at low weighted prevalence throughout the latter portion of the culture period but was not associated with mortality of either strain. These trends in growth and disease resistance for the two strains demonstrate that aquaculture performance is related to the level of disease resistance in oyster strains, salinity of water in growing areas and virulence of P. marinus.     

Experimental pathogenicity of reovirus 13p2 for juvenile American oysters Crassostrea virginica (Gmelin) and bluegill fingerlings Lepomis macrochirus (Rafinesque)

Abstract. A viral agent (13p₂), isolated from clinically normal juvenile American oysters ( Crassostrea virginica ) and characterized as a new serotype of reovirus, was tested to determine if it could replicate and produce disease in experimental juvenile oysters. Because the virus replicated well in the bluegill fry (BF-2) fish cell line, fingerling bluegills Lepomis macrochirus were included in the pathogenicity experiments. Exposure of oysters to the 13p₂ virus in ambient seawater resulted in no significant mortality and no increased virus titres or histological lesions. Virus particles were not observed in tissues of exposed oysters when examined with transmission electron microscopy (TEM). Intraperitoneal inoculation of the 13p₂ virus into bluegills resulted in 44% mortality associated with a focal necrotic hepatitis. Evidence of virus replication, when evaluated histological or by virus titration, was observed in 94% of 32 inoculated fish. Samples of infected livers examined with TEM revealed typical cytoplasmic arrays of 13p₂ virus particles in affected hepatocytes. Rising virus titres and hepatic lesions also occurred in bluegills exposed to water containing the 13p₂ virus. These results indicated the natural host of the 13p₂ virus was not the American oyster, but that it was a significant pathogen for at least one fish species.     

Growth of triploid oyster, Crassostrea madrasensis (Preston)

The performance of I and II meiotic triploids and control oysters (Crassostrea madrasensis) reared at Tuticorin Bay was compared to determine if the improvements in the growth of edible oysters were additive to faster growth in triploids. After a grow‐out period of 12 months, both mean whole weights and shell heights were in order I meiotic triploid>II meiotic triploid>control. Mean whole weights and shell height of different oyster lines were all significantly different (P<0.05). On an average, larger morphological traits indicated that growth improvements from triploids were additive, and throughout the study triploid oysters maintained faster growth rate than their diploid siblings. Condition index and adductor muscle diameter of both triploids were higher than those of control.     

Growth of the oyster Crassostrea corteziensis (Hertlein, 1951) in Sonora, Mexico

This is the first evaluation of growth and survival of spat of the Cortez oyster Crassostrea corteziensis (Hertlein) produced under controlled conditions in a coastal area in the state of Sonora, Mexico for aquaculture purposes. A suspended culture technique, used for the Pacific oyster C. gigas, was used. The Cortez oyster has an isometric shell growth during the first 13 months, reaching 71.3±1.9 mm length, 52.6±1.3 mm thickness and 25.1±0.8 mm width. Allometric growth was found between total weight and length, thickness and width (survival was 70%). The relationships between particulate organic, inorganic material, chlorophyll a and environmental parameters with growth are described. Growth rates of C. corteziensis were affected by temperature with retardation at less than 18°C. For aquaculture purposes, it is recommended that spat be sowed after winter, and oyster harvest occur at the end of autumn. According to the von Bertalanffy equation, Cortez oysters would reach the traditional exploitation size of 65 mm (mean length) at harvest. Finally, the results of this study have shown that C. corteziensis is a good candidate for aquaculture projects in this region.     

止锚湾浅海褶牡蛎养殖问题的探讨

将褶牡蛎的生物特点和养殖褶牡蛎所需要的环境条件与我省止锚湾浅海的自然环境条件进行比较,并以1992～1994年在该海区进行的现场调查数据为依据设计了养殖方法,同时进行了效益分析,从而提出了褶牡蛎在止锚湾浅海养殖的可行性和必要性。     

基于微卫星标记整合长牡蛎遗传图谱

为了提高长牡蛎遗传图谱上的微卫星标记密度,实验采用6个家系图谱间的共有微卫星标记作为锚定标记,构建了长牡蛎的整合图谱.该整合图谱共有161个微卫星标记,覆盖10个连锁群,图谱长度和平均间距分别为615.4和3.8 cM.各连锁群的标记数介于10 ～ 24个之间,连锁群长度为47.3～73.3 cM,是目前密度最高的长牡蛎微卫星图谱.不同作图家系连锁群上的标记分组保持一致,但标记顺序出现差异,可能与长牡蛎自然群体中存在大量的染色体重排现象有关.结果表明,该图谱可以为今后长牡蛎的遗传育种研究提供新的遗传工具.     

Characterization of haemocytes of the Indian edible oyster, Crassostrea madrasensis (Preston)

The haemocytes of the Indian edible oyster Crassostrea madrasensis were characterized using light and electron microscopy. The light microscopic study was conducted by staining a monolayer of the haemocytes with Geimsa. Cells without granules and with a large nucleus occupying much of the cytoplasmic area were grouped as hyalinocytes. Those with lesser amounts of basophilic cytoplasmic granules were characterized as semigranulocytes and those with large amounts of a mixture of acidophilic and basophilic granules were termed as granulocytes. Ultrastructural studies also revealed the presence of three types of haemocytes. Scanning electron microscopic studies were used to study the spreading behaviour of the haemocytes. Cytochemical studies revealed the presence of acidphosphatase, peroxidase and prophenol oxidase in the cells.     

Genetic improvement of the Pacific oyster Crassostrea gigas (Thunberg) in Australia

The Pacific oyster industry in Australia is derived from importations from Japan in the late 1940s and early 1950s to Tasmania and is almost completely hatchery based. This makes it a good target for developing and deploying genetically improved strains. An allozyme survey comparing hatchery stocks with self‐recruiting Tasmanian stocks and with two collections from Japan found abundant variation and no significant evidence of allele loss. The subsequent selection programme (initiated in the summer of 1996/97) had several strands. We wanted to take advantage of the increased power that marker‐assisted selection could bring and, therefore, needed to develop a linkage map and isolate flanking markers around quantitative trait loci (QTLs). Several types of markers (allozymes, microsatellites and AFLPs) were used, and single‐pair crosses were set up; QTLs have been detected. Conventional selection programmes, one based on mass selection and one on family selection, have been established. Triploid Pacific oysters produced via chemical means have been available for several years, but rates of triploidy achieved by such means are usually less than 100%. In 1999, we will assess whether our tetraploid 2‐year‐old broodstock can be crossed with diploids to give 100% triploid offspring.     

Growth and survival of the Calafia mother-of-pearl oyster Pinctada mazatlanica (Hanley 1856) under different sequences of nursery culture–late culture at Bahía de La Paz, Baja California Sur, México

The Calafia mother‐of‐pearl oyster, Pinctada mazatlanica (Hanley), and the Rainbow nacre shell, Pteria sterna (Gould), represent an important resource for México because of their potential in pearl production. The present work deals with the effect of different sequences of nursery culture‐late culture on growth and survival of P. mazatlanica, from September 1993 to October 1994. The collected spat presented two main size groups: small (mean shell height of 7 mm), and large (13 mm). They were arranged into four experimental batches for each size group at a constant stocking density of 40–45 juvenile pearl oysters per Nestier cage. Three batches remained in nursery culture for 2, 4 and 6 months respectively, after which they were transferred to late culture in rail cages. A control group remained in nursery culture for 12 months. Growth was evaluated monthly and compared through anova and HSD Tukey tests. In addition to the shell height, width, depth (mm) and weight (g), data of shell volume (height × width × depth, in mm³) was also introduced to estimate and compare growth among the experimental groups. Mortality was estimated by counting the dead specimens every month and obtaining the percentage from a 100% initial survival at the start of the experiment. The juveniles showed different responses to the change from nursery culture to late culture; the level of each response varied significantly among the experimental groups at the end of the study. It seemed that a 6‐month period for nursery culture was propitious for P. mazatlanica.     

葡萄牙牡蛎的净化及其低温贮藏研究

为保证生食葡萄牙牡蛎(Crassostrea angulata)的安全,在贝水比1∶10、水交换率3次/h、水温28℃、盐度28的条件下,采用流水式紫外线杀菌方法净化48 h,检测净化前后13个卫生指标的变化;净化后的牡蛎在冷藏(4℃)和冷冻(-18℃)两种条件下贮藏7 d以上;检测贮藏过程中菌落总数、大肠菌群和主要营养成分的变化。结果显示,经过48 h净化,菌落总数和大肠菌群含量均大幅下降,重金属、化学污染物等指标变化不大;两种贮藏条件下,菌落总数和大肠菌群都呈先下降再上升的变化,但始终处于安全水平内;蛋白质、脂肪、糖原含量缓慢下降,乳酸含量快速上升。研究表明,葡萄牙牡蛎在紫外线杀菌净化48 h后,至少在冷藏6 d或冷冻11 d内可以保证食品安全。     

Application of sperm cryopreservation in selective breeding of the Pacific oyster,Crassostrea gigas (Thunberg)

The robustness of Pacific oyster, Crassostrea gigas (Thunberg), sperm cryopreservation in the context of selective breeding based on family lines was investigated. Irrespective of egg density, high fertilization success was achieved with cryopreserved sperm when sperm:egg ratios of 1000:1 to 10 000:1 were used. Variation among replicate runs on the same oyster batches was minimal, indicating that cryopreservation and larval rearing procedures were repeatable. Twenty independent single male–female crosses were made to assess the utility of cryopreserved sperm in selective breeding. The fertility of unfrozen sperm was generally a poor predictor of cryopreserved sperm fertility. Based on D‐larval yields, 17 of the 20 crosses were likely to yield adequate spat for selective breeding (>10⁵ D‐larvae from 1 million eggs), two were marginal (5 × 10⁴ D‐larvae) and one was inadequate (4 × 10³ D‐larvae). An alternative fertilization strategy to improve D‐yield from a given number of sperm was then tested. Fertilizing 10 million eggs at a sperm:egg ratio of 200:1 increased the total D‐yield when compared with fertilizing 1 million eggs at a sperm:egg ratio of 2000:1 for the same male–female pair. We conclude that, despite wide variation in fertility, cryopreserved sperm is useful for family production.