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.     

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.     

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.     

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.     

Studies on triploid oysters in Australia: effect of initial size on growth of diploid and triploid Sydney rock oysters, Saccostrea commercialis (Iredale & Roughley)

In a 2-year grow-out trial, triploid Sydney rock oysters, Saccostrea commercialis (Iredale & Roughley), from two initial size grades grew faster (in terms of both mean whole weight and shell height) than the equivalent initial size grades of sibling diploids (P < 0.05). Small size grade triploids caught up with and had significantly heavier (P < 0.05) final whole weights than large size grade diploids after a 2-years grow-out period. The initial size grade had a significant effect on final mean whole weight and shell height for both ploidy types. After the 2-years grow-out trial, the final mean whole weights (but not shell heights) of small and large diploids (35.8 ± 0.6 g and 39.4 ± 0.5 g, respectively) were significantly different (P < 0.05). Small and large triploids grew at a similar rate for the first 18 months despite the significantly (P < 0.05) heavier final mean weight of large grade triploids (48.4 ± 0.8 g and 61.2 ± 0.7 g, respectively). The effect of the initial size grade on subsequent growth of both diploid and triploid oysters which was demonstrated in the present study is of significant commercial value to hatchery and nursery operators as well as growers of single seed oysters. In addition, small-grade triploids appeared to be more valuable in terms of potential growth rate than all diploid grades. There was no significant difference in the final percentage triploidy between small and large grade triploids. A large proportion of diploid/triploid mosaicism was detected in adult oysters.     

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.     

Development of a diagnostic test for Mikrocytos roughleyi, the aetiological agent of Australian winter mortality of the commercial rock oyster, Saccostrea commercialis (Iredale & Roughley)

Abstract. Diagnosis and epidemiology of Mikrocytos roughleyi , the aetiological agent of Australian winter mortality of the rock oyster, Saccostrea commercialis (lredale & Roughley ), has been hampered by the small size of the pathogen and the low intensity of natural infections. Inoculation of experimental oysters with filtered, unpurified tissue homogenate from oysters infected naturally produced experimental infections after 22 weeks but hyperinfection was not successful. DNA was extracted from naturally infected, experimentally infected and uninfected oysters, then oligonucleotide primers were used in polymerase chain reactions to provide the basis for a molecular bioassay. One pair of primers produced an amplicon of 680 base pairs in length that was correlated directly to the presence of M. roughleyi in oyster tissues. The assay was able to detect a single organism of M. roughleyi in 400 host cells.     

Effects of glycopeptides on development,growth and non‐specific immunity of pearl oyster Pinctada fucata (Gould)

The effects of glycopeptides, prepared from pearl oyster Pinctada fucata, on embryonic development, larval and juvenile growth and adult non‐specific immunity of P. fucata were investigated in this study. Glycopeptides had a pronounced stimulatory effect on embryonic development and larval and juvenile growth of P. fucata. enhancing with increased glycopeptide concentrations. All of haemocytes, phagocytosis, aggregation, serum microbiostatic activity and bacteriocidal activity all showed significant increases after 60‐day feeding, relative to unfed controls. The major conclusion is that glycopeptides had a pronounced stimulatory effect on the non‐specific immunity of pearl oysters.     

Ingestion and digestion of 10 species of microalgae by winged pearl oyster Pteria sterna (Gould, 1851) larvae

Ten species of microalgae were tested for ingestion and digestion in Pteria sterna larvae using epifluorescence microscopy to choose an appropriate diet. An experiment was conducted using 2, 4, 5 (straight-hinge) 10 and 22 (umbo stage) day old larvae. Larvae were stocked in 150 ml flasks at 30 ml⁻¹ and fed 100,000 algal cells ml⁻¹ of each species individually. Larvae were fed for 1 h and then were observed under the microscope to detect ingestion; larvae were then sieved and placed in flasks containing filtered seawater and were observed after 1 and 2 h to analyse digestion for the microalgae ingested. Out of the 10 species administered, only Nannochloris sp., Pavlova lutheri and Isochrysis aff. galbana (T-ISO) were ingested, and only the last two species were digested. No ingestion of Phaeodactylum tricornutum, Chaetoceros muelleri, Ch. calcitrans, Thalassiosira weisflogii, Dunaliella salina, Tetraselmis tetrathele and T. suecica was evident at any stage of larval development tested. Only T-ISO and P. lutheri should be used for larval rearing of P. sterna until other species are identified as suitable (ingested and digested) for this species.     

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.