Mortality in single‐pair mating families of QX disease‐resistant and wild‐type Sydney rock oysters (Saccostrea glomerata)

QX disease causes mass mortalities among Sydney rock oysters (Saccostrea glomerata). To overcome commercial production losses, Industry & Investment NSW has been developing mass selected QX disease‐resistant breeding lines since 1997. This breeding programme has significantly reduced QX‐associated mortality in the Lime Kiln Bar (LKB) breeding line relative to non‐selected, wild‐type (WT) oysters. The current study assessed mortality in families produced by single‐pair mating between LKB and WT oysters. When these families were grown in a QX disease‐prone area, the progeny of LKB × LKB crosses had significantly lower mortality compared with LKB × WT or WT × WT families. Mortality in the different crosses was associated with infection by sporulating Marteilia sydneyi, the parasite responsible for QX disease. Overall, the study identified a strong association between parentage and mortality resulting from QX disease.     

Evaluation of the progeny of the fourth‐generation Sydney rock oyster Saccostrea glomerata (Gould, 1850) breeding lines for resistance to QX disease (Marteilia sydneyi) and winter mortality (Bonamia roughleyi)

The Sydney rock oyster breeding programme began in 1990 and initially focused on faster growth and resistance to winter mortality disease (WM, Bonamia roughleyi) before including QX disease (QX, Marteilia sydneyi) resistance in 1997. Four generations of oysters have now been exposed to these diseases at three sites in the Georges River, NSW, Australia (Lime Kiln Bar, LKB; Woolooware Bay, WB; and, Quibray Bay, QB). Non‐selected control oysters and lines developed at each site (Line 1 – LKB, Line 2 – WB, Line 3 – QB) were placed at the three sites alongside a new QX‐resistant line (Line 4) bred from QX survivors in other NSW estuaries. Line 1 oysters grew to a marketable size (>50 g) in two years with minimal losses following QX disease outbreaks: 28% mortality compared with 97% in controls. Losses in Line 3 oysters selected for WM resistance at QB were reduced by more than half (23% versus 52%). Line 2 performed best at WB, but this line showed excellent resistance to QX at LKB and WM at QB, indicating that breeding for resistance to both diseases is effective. However, selection of oysters for QX did not confer resistance to WM and vice versa.     

Evaluation of the progeny of third-generation Sydney rock oyster Saccostrea glomerata (Gould, 1850) breeding lines for resistance to QX disease Marteilia sydneyi and winter mortality Bonamia roughleyi

A breeding program for Sydney rock oysters Saccostrea glomerata (Gould, 1850) has been selecting oysters for resistance to QX disease (Marteilia sydneyi) and winter mortality (Bonamia roughleyi) for three generations at three sites in Georges River, New South Wales, Australia. The experimental sites are located at the upper, middle and lower reaches of oyster growing areas in the estuary. QX disease mainly occurs in the middle and upper reaches and is most severe at the latter. Winter mortality on the other hand occurs mainly at the lower and middle reaches and is most severe at the former. Progeny of third‐generation Sydney rock oyster breeding lines were evaluated for resistance to both QX disease and winter mortality against a non‐selected control. Line 1, selected for QX disease resistance at the upper estuary site, had excellent resistance to one season of exposure to disease, but suffered high mortality during the second season of exposure. However, these oysters had already reached market size of 50 g whole weight, with low mortality at 2 years of age, before the second episode of QX disease. Line 2 showed good improvement in resistance to both diseases, whereas Line 3, was the most resistant to winter mortality. Selection for resistance to QX disease did not appear to confer resistance to winter mortality and the converse also applied.     

Experimental deepwater culture of the Sydney rock oyster (Crassostrea commercialis=Saccostrea cucullata): I. Growth of vertical clumps of oysters (‘ren’)

Japanese style ren, consisting of scallop shells encrusted with small Sydney rock oysters and strung on vertical wires, were grown at 23 estuarine stations along the New South Wales coast for up to 21 months. Weight increments of ren were used to evaluate growth rates of oysters in non-cumulative (3–4 months immersion) and cumulative (6–21 months immersion) series. Oyster weight increased at a rate of approximately 10% per month, which was considered suitable for commercial production, but estimated labour costs to make up the ren were high. Mortality rate was 6.3% per 3–4 months immersion. Gaping oysters, through natural mortality, winter mortality, the haplosporidian parasite Marteilia sydneyi, and other causes, accounted for 4.5%. Broken oysters, resulting from fish and crab predation, accounted for 1.8%. Mortality caused by the polychaete mudworm Polydora websteri was negligible; only 1.7% of the 4.5% gapers showed the characteristic blisters.     

Shell shape and meat condition in selectively bred Sydney rock oysters,Saccostrea glomerata (Gould, 1850): The influence of grow‐out methods

The Australian edible oyster industry has been severely impacted by disease and declining yields since the 1970s. Selective breeding of Saccostrea glomerata is one measure addressing these problems by producing fast‐growing, disease‐resistant oysters. Farmers report that selected oysters have different growth characteristics than their wild counterparts using conventional grow‐out methods. This study investigated how different grow‐out methods influence commercially valuable oyster characteristics including shell length, shape, surface growth deformities and meat condition. In June 2015, selectively bred S. glomerata spat were deployed in two estuaries (Hawkesbury River and Georges River) in NSW, Australia, using three grow‐out methods (fixed trays, Stanway cylinders and floating baskets). In November 2015, oysters were transferred among grow‐out methods to test for the effects of changing grow‐out methods on oyster growth patterns. Oysters transferred from baskets to cylinders and from trays to cylinders had, on average, deeper and wider shells, a higher meat condition and fewer shell surface deformities than oysters in other grow‐out method combinations. However, these oysters were smaller than oysters not grown in cylinders. While there were some differences in growth patterns between the estuaries, overall it was the grow‐out methods that most influenced oyster characteristics. This was attributed to differences in the amount and magnitude of movement oysters experienced in the grow‐out methods, as recorded by motion sensors. This study demonstrates how grow‐out methods can be managed to achieve desired growth trajectories and therefore improve marketability among selective bred S. glomerata.     

Evaluation of progeny of fourth generation Sydney rock oyster Saccostrea glomerata (Gould, 1850) breeding lines

The progeny of four 4th generation Sydney rock oyster Saccostrea glomerata (Gould, 1850) breeding lines that were selected for fast growth were compared in a 3‐year farming experiment. Oysters of the most improved breeding line (line 2) reached market size (≥50‐g whole weight) 15 months earlier than non‐selected control oysters (3 years and 5 months). The average reduction in time to market size for oysters of all four breeding lines was 12.5 months.     

The effect of low salinity on phenoloxidase activity in the Sydney rock oyster,Saccostrea glomerata

This study tested the effects of low salinity on phenoloxidase activity in Sydney rock oysters (Saccostrea glomerata). Phenoloxidase is a key component of the immune system in S. glomerata. Previously, we have shown that decreased phenoloxidase activity is associated with susceptibility to fatal QX disease. In the current study, laboratory-based experiments were used to identify factors that contribute to decreased phenoloxidase activity. We found that exposing oysters to water collected from a QX-prone oyster growing area after heavy rain significantly inhibited their phenoloxidase activities. Similar inhibition was evident when oysters were held in oceanic water with artificially lowered salinities. These results suggest that exposure to low salinity decreases phenoloxidase activity. Field trials that exploited a natural salinity gradient in the Georges River, Sydney, supported this conclusion. The phenoloxidase activities of oysters transplanted to up-river sites that had low salinities were significantly lower than those of oysters held at seaward sites with higher salinities. All of these data implicate low salinity as a key environmental stressor that is associated with inhibition of the S. glomerata immune system.     

Parasites, pathological conditions and mortality in QX-resistant and wild-caught Sydney rock oysters, Saccostrea glomerata

Differences in the survival of QX-resistant fifth generation (QXR₅) and wild-caught (Wild-Caught) Sydney rock oysters were assessed over the QX-disease risk period in the Pimpama River, SE Queensland. Cumulative mortality of Wild-Caught oysters (31.7%) was significantly greater than QXR₅ oysters (0.0%) over the 118 days of the experiment. PCR and histological results showed that Wild-Caught oyster did not die from QX disease. Histology revealed oysters were infected with disseminating hemocytic neoplasia, a Steinhausia-like infection, a Rickettsia-like organism infecting epithelial cells of the gill, digenean flukes encysted in the gonadal tissue and a gill response to an unknown toxin. The cause of mortality is attributed to disseminating hemocytic neoplasia.     

Testing options for the commercialization of abalone selective breeding using bioeconomic simulation modelling

The genetic response and economic benefit from alternative breeding programme designs for blacklip and greenlip abalone (Haliotis rubra and Haliotis laevigata, respectively) were evaluated using a computer simulation model. Two selection criteria were investigated, one used family breeding values for liability to disease challenge test infection and the other used a direct selection of the best performing individuals across families for growth rate. Five scales of breeding programme were tested and the model predicted that if growth rate is the only selection criterion, breeding programmes of a scale using 150 families of each species each generation would result in 12–13% genetic improvement in initial generations and have the greatest beneficial economic impact on the Australian abalone industry of the options tested. The model predicts an average discounted benefit–cost ratio of 48:1, total added discounted benefit of AU$4.90 for each kilogram of abalone produced and nominal economic effect on operating income of over AU$16 million per year after 10 years. If disease resistance is the only selective breeding criterion, 100 families of each species would result in the highest benefit–cost ratio of the options tested, although some genetic gain would need to be sacrificed to reduce inbreeding to acceptable levels in this scenario. A strategy for a stand‐alone abalone selective breeding cooperative was also modelled. For a farm of current tank area yielding 100 t year^?1, participation is expected to yield over AUThe genetic response and economic benefit from alternative breeding programme designs for blacklip and greenlip abalone (Haliotis rubra and Haliotis laevigata, respectively) were evaluated using a computer simulation model. Two selection criteria were investigated, one used family breeding values for liability to disease challenge test infection and the other used a direct selection of the best performing individuals across families for growth rate. Five scales of breeding programme were tested and the model predicted that if growth rate is the only selection criterion, breeding programmes of a scale using 150 families of each species each generation would result in 12–13% genetic improvement in initial generations and have the greatest beneficial economic impact on the Australian abalone industry of the options tested. The model predicts an average discounted benefit–cost ratio of 48:1, total added discounted benefit of AU$4.90 for each kilogram of abalone produced and nominal economic effect on operating income of over AU$16 million per year after 10 years. If disease resistance is the only selective breeding criterion, 100 families of each species would result in the highest benefit–cost ratio of the options tested, although some genetic gain would need to be sacrificed to reduce inbreeding to acceptable levels in this scenario. A strategy for a stand‐alone abalone selective breeding cooperative was also modelled. For a farm of current tank area yielding 100 t year^?1, participation is expected to yield over AU$0.7 million in discounted total added production value and annual discounted returns of over AU$0.4 million per annum by year 10.     

Defining Dermo resistance phenotypes in an eastern oyster breeding population

Perkinsus marinus, the causative agent of Dermo disease, is responsible for mass mortalities and negatively impacts aquaculture production of the eastern oyster, Crassostrea virginica. Selective breeding is a viable option for Dermo disease management; however, fluctuations in natural selection pressure and environmental noise hinder accumulation of genetic gains acquired through field performance trials. The purpose of this study was to adapt and apply laboratory disease challenge methods to eastern oysters, better characterize resistance‐specific traits and assess the potential for genetic variation in Dermo resistance among distinct families within a breeding population. Two challenge experiments were conducted, one in 2014 and the other in 2015. Significant treatment (control vs. challenged) and family effects on survival (measured as per cent survival and days to death) were detected in the 2014 challenge, while overall high survival precluded the detection of a significant family effect in the 2015 challenge. An alternate measure of resistance, parasite elimination rate, was also measured in the 2015 challenge, and this varied significantly among families. Thus, both survival and the change in parasite concentration in oyster tissues over time represent Dermo resistance phenotypes that can be measured accurately with the adapted laboratory disease challenge protocol described here. The obvious next step is to incorporate the challenge protocol in eastern oyster breeding programmes to assess whether well‐defined, accurately measured, Dermo‐resistant phenotypes result in enhanced genetic improvement for this commercially important trait.     

The effect of food quality on glycogen content,the fatty acid profile and winter mortality in cultivated oyster spat (Ostrea edulis)

Winter mortality in hatchery reared oyster spat (Ostrea edulis) that received three different diets during the summer period was investigated. Oysters fed a natural type diet had a winter mortality of 18.3 ± 6.3% while oysters fed cultivated algae (a mixture of Tetraselmis suecica, Isochrysis galbana and Chaetoceros muelleri) had a mortality of 73.0 ± 9.7%. A group of oysters fed a mix between the two diets had a mortality of 54.7 ± 10.6%. Tissue samples were taken at the start of the experiment, after the summer period and after the winter period in order to determine growth and the content of glycogen and fatty acids. The glycogen content decreased for all groups during the winter but the decrease was highest in oysters fed the natural diet. This group also contained the largest variety of fatty acids, but there was no difference in the content of the essential fatty acids EPA, DPA and DHA between the groups. It is concluded that transplantation of spat to the sea in spring and early summer may reduce winter mortality since the feeding period on a more varied natural algal diet is prolonged compared to transplantation of spat later in the season.     

Reproductive condition of the pearl oyster, Pinctada imbricata, Röding, in Port Stephens, New South Wales, Australia

In an investigation of the potential for pearl production in New South Wales (NSW), Australia, changes in the physical and reproductive condition of the pearl oyster, Pinctada imbricata, were monitored for over 2 years. Using wild oysters gathered from close to the southern extent of the species' range in Port Stephens, NSW, a series of macroscopic and histological observations were made. Reproductive activity in P. imbricata was greatest from late spring to early autumn with oysters in poor reproductive condition during winter. Peaks in reproductive indices occurred in November 1998, March 1999, December 1999 and April 2000. Four indices of physiological condition were used: shell growth, byssal attachment, mantle thickness and mucoprotien layer. With the exception of the thickness and extent of the mucoprotein layer, these indices either showed little variation or the variation that occurred was not related to seasonal or reproductive changes. Changes in the mucoprotein layer were correlated with water temperature and suggest that this layer is metabolized during periods of high demand such as during gonadogenesis. Collectors deployed at two sites in Port Stephens demonstrated that spatfall occurs largely in the months of December and January following the spring and early summer peaks in reproductive activity (November 1998 and December 1999). Spatfall was not observed following the autumnal peaks (March 1999 and April 2000) in reproductive activity. Overall, reproductive patterns in P. imbricata are poorly suited to spat supply in Port Stephens. Farmers require spat in early spring (September) to allow maximum use of the ‘growing’ season (September–May). Reproductively capable oysters are not available from the wild until September and quantity of natural spatfall is too variable and occurs too late in the season (December–January). As a result, oysters are being conditioned in the hatchery in July, spawned in August and spat are supplied to farmers in mid September.     

The capacity of oysters to regulate energy metabolism‐related processes may be key to their resilience against ocean acidification

Bivalve molluscs, such as oysters, are threatened by shifts in seawater chemistry resulting from climate change. However, a few species and populations within a species stand out for their capacity to cope with the impacts of climate change‐associated stressors. Understanding the intracellular basis of such differential responses can contribute to the development of strategies to minimise the pervasive effects of a changing ocean on marine organisms. In this study, we explored the intracellular responses to ocean acidification in two genetically distinct populations of Sydney rock oysters (Saccostrea glomerata). Selectively bred and wild type oysters exhibited markedly different mitochondrial integrities (mitochondrial membrane potential) and levels of reactive oxygen species (ROS) in their hemocytes under CO₂ stress. Analysis of these cellular parameters after 4 and 15 days of exposure to elevated CO₂ indicated that the onset of intracellular responses occurred earlier in the selectively bred oysters when compared to the wild type population. This may be due to an inherent capacity for increased intracellular energy production or adaptive energy reallocation in the selectively bred population. The differences observed in mitochondrial integrity and in ROS formation between oyster breeding lines reveal candidate biological processes that may underlie resilience or susceptibility to ocean acidification. Such processes can be targeted in breeding programs aiming to mitigate the impacts of climate change on threatened species.     

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.     

Effect of ploidy on the mortality of Crassostrea gigas spat caused by OsHV‐1 in France using unselected and selected OsHV‐1 resistant oysters

The effect of ploidy on the mortality of Crassostrea gigas spat caused by the ostreid herpesvirus (OsHV‐1) genotype μVar was investigated at five sites along the Atlantic coast in France in 2011. Sibling diploids and triploids were produced using either unselected or selected OsHV‐1‐resistant oysters. No significant interactions were found between the factors of environment, genotype and ploidy at the endpoint dates. The mean mortality rates at the sites were 62% and 59% for diploids and triploids, respectively, and the two rates were not significantly different. The mean mortality rates were 33% and 32% for sibling diploids and triploids, respectively, when OsHV‐1‐resistant parents were used and 91% and 85%, respectively, when unselected parents were used. The results were confirmed through other broodstocks tested in 2013. Our study is the first to clearly show that mortality related to OsHV‐1 is similar between diploids and triploids in C. gigas when the same germplasm is used for both ploidy. Furthermore, OsHV‐1 resistance was not substantially altered by triploidization, indicating that the achieved selective breeding of diploid oysters for OsHV‐1 resistance can be translated into improved survival in triploids.     

Dual disease resistance in a selectively bred eastern oyster, Crassostrea virginica, strain tested in Chesapeake Bay

Selective breeding efforts have yielded oyster strains, Crassostrea virginica, with improved survival and resistance against Haplosporidium nelsoni (MSX); however, because of susceptibility to the oyster pathogen Perkinsus marinus (Dermo), their utility has been limited in areas where the two parasites co-occur. Dual resistance to H. nelsoni and P. marinus was achieved through four generations of artificial selection of wild Delaware Bay oyster progeny at a site in the lower York River, Virginia, USA where both diseases are enzootic. During 1993–1995, survival, growth, and disease susceptibility of third generation Delaware Bay (F₃-DEBY) oysters were evaluated at the York River site in comparison to that of similarly selected third generation James River, Virginia oysters (F₃-JR), and first generation Louisiana oysters (F₁-LA), whose parents were naturally selected in the wild for resistance to P. marinus. During 1997–1999, the performance of F₄-DEBY was evaluated at three sites in Virginia in comparison to two groups of first generation oysters whose parents originated from Mobjack Bay and Tangier Sound, Virginia where both P. marinus and H. nelsoni are enzootic. In the presence of high infection pressure from both H. nelsoni and P. marinus, the F₃-DEBY stock showed significantly higher survival and growth than either the F₃-JR or F₁-LA strain. After 15 months of deployment, 79% of F₃-DEBY, 11% of F₃-JR and 17% of F₁-LA oysters were market size (≥76.2 mm) and cumulative mortality was only 16% in F₃-DEBY as compared to 42% in F₃-JR and F₁-LA. At the termination of the study, F₃-DEBY oysters exhibited 22% lower mortality than the F₁-LA stock, which outperformed the F₃-JR stock. Relative performance in respect to disease varied considerably with sample date; however, average H. nelsoni weighted prevalence varied such that F₃-DEBY1-LA=F₃-JR and average P. marinus weighted prevalence varied such that F₁-LA3-DEBY3-JR. In the 1997–1999 trial, F₄-DEBY oysters experienced 34–61% lower mortality, greater growth rate, and consistently lower prevalence and intensity of P. marinus than either Mobjack Bay or Tangier Sound oysters. H. nelsoni prevalences were very low (<12%) in all three stocks. This is the first study to demonstrate that reduced susceptibility to both P. marinus and H. nelsoni can be achieved through selective breeding.     

Realized heritability and response to selection for shell height in the pearl oyster Pinctada fucata (Gould)

The common pearl oyster, Pinctada fucata (Gould), is the most important species that is cultured for production of marine pearls in China. Heavy mortality and the decline of pearl quality have resulted in a breeding programme being established in recent years. In this study, we conducted selective breeding for the second generation of pearl oyster P. fucata (JCS-2) by mass selection for shell height (SH) with a selection intensity of 1.614, and analysed the growth of the selected line (JCS-2) and the non-selected control line (JCC) during a 1-year grow-out period. The results show that the selected line grew faster than the control one in the SH and total weight (TW) ( P <0.05), and there were higher proportion of larger sized oysters. Coefficient of variation for SH of JCS-2 was smaller than that of JCC. The current genetic gains and realized heritability for JCS-2 averaged 16.03 ± 4.79% and 0.713 ± 0.208 at 3–15 months of age respectively. The findings indicated the selection response to faster growth for SH is markedly effective in the second generation, and there was a high correlated response of TW when selecting for SH.     

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.     

Third generation evaluation of Sydney rock oyster Saccostrea commercialis (Iredale and Roughley) breeding lines

The progeny of four breeding lines of Sydney rock oysters, Saccostrea commercialis, selected for faster growth over two generations, were compared with controls in an 18-month farming experiment. Mean whole weights of oysters from all four selection lines did not differ significantly from each other but, were significantly greater (P<0.05) than mean whole weight of controls (spat produced from two groups of non-selected oyster populations). The mean increase in weight for the selection lines over the controls of 18% (range 14–23%) after two generations of selection (third generation) compares extremely favourably with the 4% reported previously after one generation of selection.     

Studies on triploid oysters in Australia: farming potential of all-triploid Pacific oysters, Crassostrea gigas (Thunberg), in Port Stephens, New South Wales, Australia

Triploid Pacific oysters Crassostrea gigas farmed in Port Stephens, NSW had an exceptionally fast growth rate and reached a whole weight of 55 g in 13 months versus 20 months for diploids. Mortality of the triploids (24.5±2.94%) was significantly lower (P<0.05) than that of the diploids (40.0±2.26%) over the duration of the experiment (July 2002–February 2004). Unfortunately, this advantage was offset by discoloration of the meats of the triploids when they were in better condition than the diploids over summer (October 2003–March 2004). However, discoloration of meat of triploids had cleared up by April 2004 and neither did they suffer this problem from April–September 2003. The triploids also had a lower peak condition than the diploids. Oysters in peak meat condition, i.e. spawning condition, are preferred for the half shell trade in Australia and in this study, there was at least a six‐month period prior to discoloration, when the triploids were large enough and had sufficient meat condition for marketing on the half shell.