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.     

The Sydney rock oyster Saccostrea glomerata (Gould 1850) breeding programme: progress and goals

In 1990, NSW Fisheries initiated a mass selection programme in Port Stephens, NSW, with the aim of breeding faster growing Sydney rock oysters Saccostrea glomerata (Gould 1850). After two generations of selection, an average weight for age advantage of 18% (range 14–23% per breeding line) was achieved. This equates to a reduction of 3 months in the time taken to reach market size. Experiments are planned to determine how much of this 3 months advantage is additive to the 6 months advantage this laboratory has already obtained using triploid S. glomerata. A parallel set of S. glomerata breeding lines was established on the Georges River, NSW, to include selection for resistance to the protistan parasite Mikrocytos roughleyi, the causal agent of winter mortality. The programme was disrupted by the outbreak of QX disease Marteilia sydneyi, another protistan parasite, in 1994. In 1997, the breeding programme was reorganized and expanded. New lines were bred from oysters that had survived both QX and winter mortality. After one generation, a small improvement in resistance to QX has been recorded; however, the exposure of further generations to M. sydneyi will be required to confirm an increase in resistance.     

Heritability estimates for nutritional quality‐related traits of the Pacific oyster,Crassostrea gigas

Pacific oysters, Crassostrea gigas, are the most abundantly harvested shellfish in the world and are ecologically significant. The content of nutrients, including protein, glycogen, lipid, zinc (Zn), and selenium (Se), is important for oyster meat quality, but heritability estimates of such traits have rarely been reported. In this study, 64 full‐sib families were generated using a nested mating design. Finally, 18 full‐sib families, of which there were nine half‐sib families, with each containing 2 full‐sib families, were sampled for heritability estimates. The narrow‐sense heritabilities of glycogen, protein, lipid, Zn, and Se contents were 0.29 ± 0.02, 0.38 ± 0.02, 0.58 ± 0.08, 0.02 ± 0.02, and 0, respectively. Negative genetic correlations existed between both glycogen and protein content (?0.95 ± 0.004) and between lipid and protein content (?0.59 ± 0.05), whereas a positive correlation was observed between lipid and glycogen content (0.16 ± 0.06). Weak genetic and phenotypical correlations (r = 0–0.2) were observed between shell height and nutritional quality traits. These data demonstrated that glycogen, protein, and lipid content can be chosen in a selective breeding program, but glycogen and lipids cannot be selected together with protein. Furthermore, performing indirect selective breeding for quality traits by selecting traits related to growth is impossible. This study provides information for the development of breeding strategies for oyster quality traits.     

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.     

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.     

The effect of temperature on physiological energetics of a fast‐growing selective strain and a hatchery population of the Pacific oyster (Crassostrea gigas)

Growth is an important aquaculture trait, which has been one of the most important traits for genetic improvement in aquatic species. Numerous fast‐growing domesticated aquaculture organisms are being cultured, but the physiological processes underlying growth remain largely unexplored. The Pacific oyster (Crassostrea gigas) is one of the most widely farmed bivalves. In order to improve the productivity traits, a fast‐growing selective strain of C. gigas was established. In this study, we compared physiological energetics of the fast‐growing selective strain and an unselected hatchery population of C. gigas under different conditions of temperature. Components of energy gain (clearance rates, ingestion rates, absorption efficiency and absorption rates), energy loss (metabolic rates) and resulting energy budget (scope for growth) were analysed at four temperatures (15, 20, 25 and 30°C). The results indicated that selective breeding oysters had a higher energy gain compared to unselected oysters, but the selected strain and hatchery population of oysters did not significantly differ in metabolic rates at all tested temperatures (p > 0.05). The enhanced energy gain resulted in almost two times higher scope for growth in selective breeding oysters compared with unselected oysters. Physiological characteristics of C. gigas were greatly affected by temperature and showed a fluctuation with the increase in temperatures. The selective breeding oysters outperformed those of hatchery population in growth performance by an increased energy gain along with a similar energy loss. This study provided insight into physiological energetics accounting for growth improvements in fast‐growing selective strain of C. gigas.     

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.     

Heterosis analysis at early generations for complete diallel crosses in three different geographical culture populations of Sinonovacula constricta (Lamarck 1818) in Zhejiang,China

The paper reports a study focused on breeding new rapidly growing and heat‐tolerant strains in a commercially important Asian bivalve, Sinonovacula constricta. To breed a new strain of S. constricta with fast growth and high temperature tolerance, complete diallel crosses were carried out using three different geographical culture populations from Zhejiang (Z), Fujian (F) and Guangdong (G) provinces, China. Larval and spat growth and the survival and heat tolerance of spats were compared in detail. The experimental serial growth traits extracted from larval and spat measurements (shell length and shell height) indicated that there was no difference between six hybrid crosses and parental crosses at the larval stage; while at the spat stage, some differences were observed. The hybrid crosses ZF (Z♀ × F♂), ZG (Z♀ × G♂) and FG (F♀ × G♂) outperformed the parental groups in shell length at the end of intermediate culture, with lengths of 21.77 ± 0.28 mm, 21.55 ± 0.29 mm and 21.44 ± 0.20 mm respectively. For spat survival rate, the hybrid cross ZF (Z♀ × F♂) had a significantly larger value (88.89 ± 4.16%) than all the other groups, and the hybrid cross ZG (Z♀ × G♂) had a significantly larger rate than the cross GZ (G♀ × Z♂) and parental crosses ZZ (Z♀ × Z♂) and GG (G♀ × G♂). The hybrid crosses ZF (Z♀ × F♂), ZG (Z♀ × G♂) and FG (F♀ × G♂) exceeded the parental groups in heat tolerance, but there was no significant difference. The growth, survival and heat tolerance results showed that the crosses ZF, ZG and FG in this study were the best hybridization combinations for genetic improvement, which could have significant implications for the development of razor clam aquaculture. This work provides a theoretical basis for breeding new varieties (strains) that grow rapidly and are heat tolerant.     

Production of inbred larvae through self‐fertilization using oocytes and cryopreserved sperm from the same individuals after sex reversal in eastern oyster Crassostrea virginica

The eastern oyster Crassostrea virginica can change sex which makes self‐fertilization possible if sperm can be cryopreserved. In this study, small (~1 year old) and large (~2–3 years old) oysters were biopsied for sperm collection. Survival of the biopsied oysters after 1 year was 50% for small oysters and 17% for large oysters. Oocytes were collected from sex‐reversed females, and self‐fertilized with cryopreserved sperm. Of the 24 cryopreserved samples, 14 individuals had ≤1% fertility when crossed with oocytes from unrelated females, indicating that the cryopreserved sperm had reduced fertility. The other 10 individuals had a fertility of 39 ± 25% when crossed with oocytes from unrelated females (non‐selfing), but showed a significantly lower success of self‐fertilization (12 ± 16%) (P = 0.008), while aliquots of the same oocytes had a fertilization of 83 ± 11% when crossing with fresh sperm. Larvae were produced at day 3 in the self‐fertilized families (12–94% of the fertilized oocytes), and survived to eyed‐larvae stage at days 11–14. Genotyping with 9 microsatellite markers confirmed that the larvae resulted from self‐fertilization in four families. This study demonstrated the feasibility of creating self‐fertilized inbred lines of oysters by use of non‐lethal sperm collection and cryopreservation.     

Fertilization, hatching, survival and growth rates in reciprocal crosses of two strains of an African catfish Heterobranchus longifilis Valenciennes 1840 under controlled hatchery conditions

A diallelic cross between two strains [Layo strain (LS) and Noun strain (NS)] of the catfish Heterobranchus longifilis Valenciennes was carried out under controlled hatchery conditions to estimate their reproductive performance and aquaculture potential in terms of fertilizability, hatchability, survival, growth and heterosis. The average fertilization rate of all mating groups was as high as 90.4%, the fertilization rate of the purebred NS (95.2%) being significantly higher than that of the purebred LS or the reciprocal crosses (P < 0.05). The average hatching rate of all genetic groups was as high as 84.7%, the hatching rate of NS (89.4%) being significantly higher than that of the purebred LS or the reciprocal crosses (P < 0.05). The mean survival rate of all crosses from hatching up to the onset of exogenous feeding stood at 94.2%, without showing a significant difference (P > 0.05) between the crosses. During the larval rearing period, which extended from the onset of exogenous feeding up to 15 days of age, NS displayed a significantly lower growth rate (P < 0.001) than that of the purebred LS and their reciprocal hybrids. There was no significant difference (P > 0.05) in survival rate (mean 84.7%) between the four crosses at the end of the larval rearing period. During the juvenile rearing phase, the mean growth performance of all crossbreds was similar to that of the purebred LS and significantly different from that of the purebred NS (P < 0.001). The final individual weights attained by LS, NS, LS × NS and NS × LS were 3.31, 0.71, 3.97 and 3.66 g respectively. The increase in weight attained by the fast‐growing crossbred LS × NS was 562.2%, 20.4% and 8.7% more than that of NS, LS and NS × LS respectively. The survival rate of NS (57.1%) was significantly lower (P < 0.001) than that of all the rest of the crosses. The crossbreds displayed about 15.1% heterosis in mean body weight relative to the fast‐growing purebred LS. It was concluded that cross‐breeding of H. longifilis strains could be advantageous because of the hybrid vigour in the progeny.     

Siltation increases the susceptibility of surface‐cultured eastern oysters (Crassostrea virginica) to parasitism by the mudworm Polydora websteri

Mudworm (Polydora websteri) parasitism is known to result in unsightly mud blisters in eastern oysters (Crassostrea virginica), resulting in reduced product quality and increased vulnerability to illness and environmental stress. While typically a concern only for bottom‐grown oysters, an abnormal severe outbreak of P. websteri in surface‐cultured oysters in New Brunswick, Canada, was reported in 2013, along with an anecdotally reported concurrent increase in siltation. Although heavier loads of silt are reported to increase P. websteri infestations in oysters, studies report mixed effects of siltation to this regard. Here, we report the results of a field experiment testing the effect of siltation on P. websteri recruitment in surface‐grown oysters at an aquaculture site in Richibucto, New Brunswick. Live oysters of similar size were deployed at the aquaculture site and were left to collect P. websteri recruits under one of two siltation treatments (high vs. low) for 50 days. Results suggested that P. websteri recruitment correlated with metrics of oyster size (shell weight, length, width, depth and surface area). When P. websteri counts were standardized for oyster shell morphometry, P. websteri recruitment was significantly higher in oysters exposed to the high siltation treatment, accumulating approximately 1.5× as many P. websteri as oysters exposed to the low siltation treatment. Our results suggest that higher amounts of siltation on surface‐cultured oysters can result in increased rates of P. websteri parasitism. Enhanced cleaning regimes may help to alleviate the impacts of P. websteri in surface‐grown oysters, although other mitigation strategies exist.     

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.     

Susceptibility of koi × crucian carp and koi × goldfish hybrids to koi herpesvirus (KHV) and the development of KHV disease (KHVD)

Hybrids of koi, Cyprinus carpio × crucian carp, Carassius carassius and koi × goldfish, Carassius auratus, proved to be susceptible to koi herpesvirus (KHV, syn. CyHV‐3) and developed KHV disease (KHVD). While hybrids of koi × goldfish were partly resistant to mortality following infection by immersion, most koi × crucian carp hybrids died after bath infection. KHV DNA was detected in dead fish but also in all surviving animals by different polymerase chain reactions (PCRs). According to these results, hybrid crossbreeding does not seem to prevent severe losses associated with KHV in terms of inducing KHVD. The present study showed severe losses after a waterborne KHV infection of between 35% and 100% in koi × goldfish and koi × crucian carp hybrids as well as in SPF carp.     

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.     

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.     

Efficacy of a Live‐attenuated Edwardsiella ictaluri Oral Vaccine in Channel and Hybrid Catfish

This study evaluated the efficacy of an oral live‐attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish (ESC) in channel and hybrid catfish. The vaccine was delivered one time orally by feeding fish a diet coated with an attenuated E. ictaluri isolate at four doses to deliver between 4 × 10⁶ to 3.2 × 10⁷ viable vaccine cells/g wet feed. Thirty‐five days postvaccination, control and vaccinated fish were challenged with virulent E. ictaluri and mortality was examined for 30 d postchallenge. Mortality of nonvaccinated hybrids (85%) and nonvaccinated channel catfish (73%) was similar but significantly greater than all groups of vaccinated fish. In channel catfish, a trend toward increasing mortality with decreasing dose was observed. Mortality of channel catfish vaccinated with the lowest dose (26.6%) was significantly higher than fish vaccinated with the highest dose (14.1%) but similar to fish vaccinated at the intermediate doses (17.5 and 19.4%). In contrast, mortality of four doses of vaccinated hybrid catfish was similar and ranged between 10.4 and 14.0%. The data demonstrate that the attenuated E. ictaluri vaccine at all four doses tested is effective at reducing ESC‐related mortalities in hybrid and channel catfish.     

Evaluation of a new select‐breeding carp variety with high fillet quality

The morphological characters, fillet texture, and nutrient contents of the F3 generation of Barbless carp (Cyprinus pellegrini,BC) female × Heilongjiang carp (Cyprinus carpio,HLC, geographical population of common carp) male hybrid, which were named as High‐quality carp F3 (HQC F3), were studied. The main morphological characteristics of HQC F3 were between BC and HLC, but closer to the BC. The fillet amino acids and crude protein showed significant differences between HQC F3 and BC. But, only fillet crude fat showed significant differences between HQC F3 and HLC. HQC F3's fillet had significantly different with BC's on the contents of myristic acid, linolenic acid, and stearic acid, and with HLC's on the contents of linoleic acid, linolenic acid, EPA, palm oleic acid, and 20 carbon‐dilute acid. HQC F3 broodstocks were selected with high intramuscular fat and effective population size. HQC F4 were bred and showed improved intramuscular fat content compared to F3. The intramuscular fat of 2‐year‐old HLC F4 was significantly (p < 0.05) correlated with body weight/standard length. Intramuscular fat is a stable trait that can be continuously passed on to the next generation. This study will lay a foundation for further carp selective breeding study to improve fillet quality.     

Vaccination of Full‐sib Channel Catfish Families Against Enteric Septicemia of Catfish with an Oral Live Attenuated Edwardsiella ictaluri Vaccine

This study evaluated the efficacy of an oral live attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish in 20 full‐sib fingerling channel catfish families. The vaccine was delivered orally by feeding fish a diet coated with an attenuated E. ictaluri isolate. Sixty‐nine days postvaccination, control and vaccinated fish were challenged with virulent E. ictaluri and mortality was examined for 21 d postchallenge. Vaccinated fish had significantly lower mortality than nonvaccinated fish following challenge (P < 0.001). Mortality of vaccinated fish was 1.7 ± 1.4% as opposed to 47.8 ± 28.7% in nonvaccinated fish. Relative percent survival ranged from 87.7 to 100% with an average of 95.2 ± 4.0% (±SE) among the 20 families of fish. There were significant differences in mortality among families in nonvaccinated fish (P < 0.01) while there were no differences among vaccinated families of fish. Results indicate that the live attenuated E. ictaluri vaccine is effective at reducing mortality in channel catfish exposed to virulent E. ictaluri. These data demonstrate that genetic differences among healthy families of channel catfish are not major considerations in developing an effective vaccination program utilizing the oral vaccination platform described in this study.