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.     

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.     

Commercial assessment of growth and mortality of fifth-generation Sydney rock oysters Saccostrea glomerata (Gould, 1850) selectively bred for faster growth

The Sydney rock oyster Saccostrea glomerata (Gould, 1850) industry acquired hatchery-produced spat selected for faster growth for the first time in January 2004. Selectively bred and non-selected (Control) spat produced concurrently were used to compare performance when grown under commercial conditions. Spat were distributed to farmers in seven estuaries in New South Wales. Individual farmers cultivated these oysters using their own techniques and growth and mortality were recorded quarterly. At each site, the two oyster types were cultivated using the same culture method, location and density. Growth was compared when oysters were 27 months of age. At this time, selectively bred oysters were significantly larger and heavier than Control oysters. The same result was obtained when oysters were compared at the point in time when selectively bred oysters had reached 50 g at each site. No significant difference was found for cumulative mortality between the selectively bred oysters and Control oysters across all sites. However, the seven sites had significantly different levels of cumulative mortality. Overall, the performance of selectively bred oysters was superior to the Control oysters and selectively bred oysters reached the 50 g bench mark within 29.3 months when averaged across all sites.     

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.     

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.     

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.     

Growth and mortality of sibling triploid and diploid Sydney rock oysters, Saccostrea glomerata (Gould), in the Camden Haven River

Growth and mortality of sibling triploid and diploid Sydney rock oysters, grown at two tidal heights, three stocking densities and three different sites on the Camden Haven River estuary, were compared. Triploids grew faster than diploids. The majority of the growth rate difference occurred after the oysters reached 2 years of age. By 3.2 years, triploids were 49% heavier than diploids. Oysters grown intertidally were larger than those grown subtidally. Oysters stocked at the lightest density of 2 L per basket grew to a significantly larger size than baskets stocked at 3 or 4 L. Growth rates were significantly different at the three sites. Growth was reduced the further the oysters were from the mouth of the estuary. There was no significant difference in mortality between ploidy level, but there were differences between tidal heights, stocking densities and sites. Intertidal oysters suffered higher mortality than subtidal. Oysters stocked at the highest density had a significantly higher mortality than the lowest density; neither of these was significantly different from the medium density. The site closest to the mouth of the estuary had significantly lower mortality than the middle and upper estuary sites.     

Protein markers of Marteilia sydneyi infection in Sydney rock oysters, Saccostrea glomerata

Marteilia sydneyi is the causative agent of QX disease in Sydney rock oyster, Saccostrea glomerata . It is responsible for disease outbreaks among oysters that occur during summer and can result in up to 95% mortality. QX disease has significantly decreased S. glomerata production in some areas of Australia's eastern seaboard over the past 30 years. Marteilia sydneyi sporulates in the digestive gland of oysters leading to complete disorganization of the infected tissues. The current study used proteomics to identify potential molecular markers of sporulating M. sydneyi infection during a field trial undertaken in the Georges River, Sydney, between December 2006 and May 2007. Early stages of M. sydneyi infection were detected by polymerase chain reaction, whilst cytological examination was used to identify sporulating M. sydneyi in the gut. Protein expression in oyster haemolymph was assessed during the M. sydneyi infection period by two dimensional electrophoresis. Proteome maps identified significant differences in the expression of four proteins in oysters with sporulating M. sydneyi infections.     

Experimental deepwater culture of the Sydney rock oyster (Crassostrea commercialis = Saccostrea cucullata): II. Pontoon tray cultivation

Pontoons made from plastic pipe were tested as an alternative to racks for deepwater culture of the Sydney rock oyster. The growth and mortality of oysters permanently suspended in water on trays beneath floating pontoons were compared with oysters on trays in an intertidal zone. For both culled spat (30–31 g whole oyster weight) and seconds oysters (37–39 g) beneath pontoons the growth rate, measured by weight increases, was three times that of oysters on intertidal racks over a 5-month period. Mortality (from unknown causes) was higher beneath the pontoons. The mean mortality of spat oysters was 40% compared with 24% on the intertidal trays, and for seconds oysters was 51% compared with 34%.     

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.     

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.     

Sydney rock oyster, Saccostrea commercialis (Iredale & Roughley), filtration of shrimp farm effluent: the effects on water quality

Shrimp pond effluent water can contain higher concentrations of dissolved nutrients and suspended particulates than the influent water. Consequently, there are concerns about adverse environmental impacts on coastal waters caused by eutrophication and increased turbidity. One potential method of improving effluent water quality prior to discharge or recirculation is to use bivalves to filter the effluent. In this study, we examined the effects of the Sydney rock oyster, Saccostrea commercialis (Iredale & Roughley), on the water quality of shrimp pond effluent. Effluent from a shrimp farm stocked with Penaeus japonicus (Bate) was pumped directly into 34-L tanks stocked with different densities of oysters. Combinations of live and dead oysters were used to test the effects of three different densities of live oysters (24, 16 and 8 live oysters per tank). The concentrations of total suspended solids, the proportion of organic and inorganic matter, total nitrogen, total phosphorous, chlorophyll a and the total number of bacteria in the pond effluent water were determined before and after filtration by oysters. The oysters significantly reduced the concentration of all the parameters examined, with the highest oyster density having the greatest effect. Shrimp pond effluent contained a higher proportion of inorganic matter (72%) than organic matter (28%). The organic component appeared to be mainly detritus, with chlorophyll a comprising only a minor proportion. Filtration by the high density of oysters reduced the effluent total suspended solids to 49% of the initial level, the bacterial numbers to 58%, total nitrogen to 80% and total phosphorous to 67%. The combined effects of settlement and oyster filtration reduced the concentration of chlorophyll a to 8% of the initial effluent value.     

Reducing the impact of environmental variation in a growth rate improvement program for the Sydney rock oyster Saccostrea commercialis

A culture method (‘slats’) being used to compare Sydney rock oysters (Saccostrea commercialis) for growth rate selection was further evaluated. In this method, oysters are affixed to plastic slats in a matrix pattern. The presence of non-random positional effects associated with an oyster's location within the tray compartment and the impact of their removal was investigated in terms of row and column effects and band effects. Within each tray compartment, oysters were assigned to different ‘bands’ according to their distance from the side of the compartment. The effect of the presence or absence of outliers in the estimation of row, column and band effects was also investigated.The growth rate of oysters was found to be sensitive to apparently small environmental differences. There were substantial and statistically significant row, column and band effects for two of the three growing sites. These non-random positional effects were effectively removed by adjusting individual oyster weights on the basis of row and column means, irrespective as to whether these means had been estimated in the presence or absence of outliers. Data adjusted on the basis of band means tended to retain some substantial row and/or column effects. There was little, if any, additional advantage in adjusting the row and column adjusted data for band effects.     

Molecular diagnosis of Marteilia sydneyi (Paramyxea) in Sydney rock oysters, Saccostrea commercialis (Angas)

Abstract. To prepare a diagnostic test, the polymerase chain reaction (PCR) with universal primers was used to amplify the internal transcribed spacer 1 (ITS1) of Marteilia sydneyi rDNA. A radiolabelled probe for M. sydneyi comprising putative ITS1 rDNA gave a strong signal for genomic DNA from M. sydneyi and no signal for host genomic DNA in Southern blots. A PCR to detect M. sydneyi used a forward primer designed from an internal site within the sequenced region and a reverse universal primer. It amplified a 650-bp fragment of M. sydneyi DNA when a combination of host and parasite DNA, extracted from an infected oyster, was used as the template.     

Experimental feeding of Sydney rock oysters (Crassostrea commercialis = Saccostrea cucullata): I. Optimum particle sizes and concentrations

Oysters removed cornflour particles of 0–4 μm diameter at a rate significantly different (5% level) from the controls. For 4–6 μm particles the difference was significant at the 10 but not the 5% level. There were no significant differences in removal rates for 6–10 μm particles. Particles less than 5 μm in diameter seemed optimum for ingestion.The optimum concentration for riceflour particles less than 5 μm in diameter was approximately 2 mg/l; at this concentration there was maximum faecal production and minimum pseudofaecal production. The changeover point (50% producing pseudofaeces) was approximately 18 mg/l; above this concentration pseudodefaecation steadily increased until 35 mg/l, when pseudofaeces (as well as faeces) were being produced by 100% of the oysters. Similar results were obtained for riceflour, starch gluten effluent and active dried yeast, but the ranges were different.     

Experimental deepwater culture of the Sydney rock oyster (Crassostrea commercialis): V. Commercial raft trials

Four large rafts (6.4 × 4.3 m), each costing $ A 543 totally equipped and holding 600 kg of trayed layers of oysters, were sited in New South Wales estuaries during 1977 and 1978. Small, single oysters (spat at 25–29 g whole weight or seconds at 29–40 g) were grown to restaurant (plate) grade (40–67 g).Sixteen crops were harvested, usually after three months growout. None showed evidence of overspatting (attachment of young oysters to the crop). Growth rates (measured as oyster weight increases) ranged from 5 to 30% per month and averaged 19%. In four of the crops mortality was high (12, 22, 22 and 90%) due to parasites, predators or wave action. In the remaining 12 crops natural mortality averaged 6% total for the growout period. The average actual wholesale value of the crops was $ A 422 and this could be raised to an estimated wholesale value of $A 527 per crop by utilising pregrowout grading and harvest date selection. Several husbandry practices are recommended. (1) Grading oysters mechanically before growout to produce more uniform crops. (2) Covering top trays to prevent fish attacks and losses through waves. (3) Deciding optimum harvest date by inspection after three months growth.     

Experimental deepwater culture of the Sydney rock oyster (Crassostrea commercialis): III. Raft cultivation of trayed oysters

Sydney rock oysters, normally intertidal, were submerged below rafts in vertical stacks of 15 oyster trays extending 2 m deep. Growth rates and mortality were not good or economically encouraging. The best growth was from small culled spat (43 whole oysters/kg) to large seconds (28/kg), an increase of 59% in 9 months. The minimum mortality was 52%. Fouling growths of barnacles, tunicates, sponges and hydroids were restricted by placing experimental trays on top of the raft for several days to dry out. Compared with controls, this resulted in increased oyster growth in experimental trays during the next 6 months. Oyster mortality and incidence of mudworm blisters (resulting from the polychaete Polydora websteri) were similar in both control and experimental trays during this period. For improved growth of trayed submerged oysters the optimum vertical distance between trays and the optimum density of oysters on trays need to be determined.     

Experimental deepwater culture of the Sydney rock oyster (Crassostrea commercialis): IV. Pilot production of faft oysters

Rafts carrying five permanently submerged oyster trays were moored in nine New South Wales estuaries during 1976–1977 to determine how rapidly culled spat oysters (25–29 g whole weight) or seconds (29–40 g) could be grown into first grade (plate) oysters (40–67 g).In the first series of trials results were obtained from seven localities after 3 months summer growth. The average weight increase was 51%; mortality averaged 17%. A further series of trials were conducted for 3 months in autumn at four localities. Weight increases averaged 40% and mortality 8%.The growth rates calculated from weight increases were two to three times greater than those expected from normal intertidal culture traditional of Australia. The estimated time to convert spat to plate oysters was 4 months for summer and autumn crops.     

Experimental deepwater culture of the Sydney rock oyster (Saccostrea commercialis). VI. Commercial oyster cage systems

A growout cage that retains oysters, excludes predators and reduces maintenance is described; it is cut from 200 mm (8 in) steel reinforcing mesh as kitset units and then hot dip galvanised. Each cage is 2.2 m long, 0.8 m wide and 1.0 m deep (88 × 22 × 40 in) and contains five floors which are 203 mm (8 in) apart. The floors and the walls are covered with 19 mm ($\text{3}\text{4}$ in) galvanised wire mesh, or high density polyethylene plastic mesh. These cages may be slung from rafts, fitted with individual flotation or lowered to the bottom with a marker buoy; they weigh approximately 70 kg (154 lb) empty and about 220 kg (484 lb) when harvested. Each cage crop consists of approximately 2.5 bags totalling about 150 kg (330 lb), and in good growing areas three to four crops have been harvested from each cage per year. In two such crops culled spat (22–29 g whole wet weight) were converted to plate (first grade) oysters (40–67 g); in the other two, seconds (29–40 g) were converted to plate oysters. Growth rates, stocking densities and mortality data are given for 11 crops.     

Studies on triploid oysters in Australia: Evaluation of cytochalasin B and 6-dimethylaminopurine for triploidy induction in Sydney rock oysters Saccostrea commercialis (Iredale and Roughley)

Naturally spawned Sydney rock oysters Saccostrea commercialis (Iredale and Roughley),were used to determine the appropriate stage of development for inducing triploidy and to compare the effectiveness of cytochalasin B (CB) and 6-dimethylaminopurine (6-DMAP) in dose-optimization trials. Induction should commence at 50% first polar body (PB1) extrusion in eggs (approximately 17-19 min post-fertilization at 25^oC). By day 5 the highest triploidy percentage and yield (number of triploid larvae per 100 fertilized eggs) were achieved in the ranges of 0.75-1.5 mg CB 1^-1 (1.6-3.1 μm CB)or 200-400 μm 6-DMAP (32.6-65.3 mg 6-DMAP l^-1). However, CB treatment resulted in greater survival and triploidy percentage than 6-DMAP in Sydney rock oysters.