Phenotypic trait of Crassostrea hongkongensis♀× C. angulata♂ hybrids in southern China

The Hong Kong oyster (Crassostrea hongkongensis) and the Portuguese oyster (Crassostrea angulata) are important aquaculture species in southern China. To understand the potential feasibility of hybridization between the two species, two‐by‐two factorial cross‐experiments were conducted in Shenzhen (Guangdong province). An asymmetry in fertilization was revealed: C. hongkongensis eggs could be fertilized by C. angulata sperm, but the reciprocal cross was not possible. The fertilization and hatching success in the C. hongkongensis females × C. angulata males (HA) cross was lower than the two intraspecific crosses. Moreover, hybrid larvae had a slower growth rate than those from parental progenies due to genome incompatibility. However, the size of the spat and adult growth were greater than those of the two parental progeny, with obvious heterosis during the grow‐out stage. The survival advantage of hybrids was examined at all times. The surprising finding was that all hybrids were completely fertile, meaning they could produce normal and functional gametes. Genetic analysis by molecular markers was applied to confirm that the HA spat were true hybrids. Our results revealed that the interspecific hybridization between C. hongkongensis and C. angulata is completely successful in one direction. Furthermore, hybrids are viable, fast growing and completely fertile, which provides a promising method for the genetic improvement of oysters as a new aquaculture stock in southern China.     

Parentage assignment in hybrid abalones (Haliotis rufescens × Haliotis discus hannai) based on microsatellite DNA markers

Parentage analysis in aquaculture determines genealogical relationships between broodstock and progeny when the parents are unknown. Thus, parentage analysis is a useful tool to establish pedigree reports in molecular‐assisted selection programs. Here, we evaluated 10 heterologous microsatellite markers for parentage assignment in abalone hybrids produced from 43 abalone broodstocks of red abalone (Haliotis rufescens) and Japanese abalone (H. discus hannai). The allele frequencies, exclusion probabilities and broodstock contributions were calculated using CERVUS, PAPA and GERUD software. The polymorphic information content (PIC) values showed that most of the microsatellite loci were highly informative (>0.7) and more than 90% of parentage assignment was possible with a minimum of 5–6 microsatellite markers. Parentage assignment for hybrid and pure‐red progeny showed a better performance than pure‐Japanese progeny. This result could be due to the high level of allele loss in the parental genotypes. In addition, results indicated that only two sires contributed over 80% and 90% of red and hybrid progenies, respectively. This study gives a new molecular tool to support marker‐assisted selection in abalone hybrids produced in Chile.     

Survival and growth of reciprocal crosses between two stocks of the Hong Kong oyster Crassostrea hongkongensis (Lam & Morton, 2003) in southern China

The Hong Kong oyster, Crassostrea hongkongensis, is one of the most economically important and well‐known oyster species in southern China. To explore the possibility of improving the performance of the Hong Kong oyster, complete diallel crosses between two oyster stocks, the Zhuhai stock (Z: fast growth line F4) and the Maowei Sea stock (M: resistant line F3), were conducted using pooled gametes. Three replicates, each consisting of two pure stocks (ZZ and MM) and two reciprocal crosses (ZM and MZ), were successfully generated. High fertilization rate and hatching level were observed among all the experimental groups, suggesting that there was no sperm–egg recognition barrier between geographic populations. Reciprocal crosses had higher survival rate compared to the pure stock crosses and the rate of survival increased with progeny growth. Growth heterosis became obvious both in larval and adult stages, and was primarily influenced by the egg origin and mating strategy at the larval stage. Also, the phenotypic traits of all progeny differed amongst the culture sites, suggesting a significant environmental effect. The Zhuhai site was more suitable for oyster aquaculture than the Maowei Sea site. Thus, our results demonstrated that crossbreeding between Zhuhai and Maowei Sea stocks of Hong Kong oyster to produce superior heterosis represents a promising means to improve the fishery yield of this species in southern China.     

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.     

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

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

Genetic parameters of growth and survival in the Pacific oyster Crassostrea gigas

The Pacific oyster Crassostrea gigas is a representative bivalve mollusc that is widely cultured in the world and is the largest molluscan group cultured in China. In order to assess the feasibility of improving survival of C. gigas through genetic selection, the heritability and genetic correlations for growth and survival traits between different life stages were examined. Genetic parameters were estimated based on intraclass correlations of 49 full‐sib families (29 half‐sib families) in larvae (4 and 20 days after fertilization) and spat (140 days after fertilization) stages. The heritability for growth traits in larvae and spat was 0.30–0.86 and 0.53–0.59, respectively, and varied with ages. The heritability of survival was low in larvae (0.13 ± 0.05 and 0.17 ± 0.04, respectively for 4 and 20 days after fertilization) but medium (0.39 ± 0.07) in spat, suggesting that selection for increasing spat survival was feasible. The genetic correlation between growth traits within age was medium to high and positive (ranging from 0.47 to 0.96, respectively, between shell length (SL) and shell height (SH) at 20 days and between SL and SH at 140 days after fertilization), suggesting that selection to improve single growth trait will cause positive response in another growth traits in C. gigas. The genetic correlations between survival and growth traits at 140 days were low but positive (ranging from 0.23 to 0.27, respectively, between survival and SH and between survival and SL at 140 days after fertilization), suggesting that selection for survival may not have a negative response in growth. Overall, this study suggests that survival traits should be taken as improving target of next selection breeding programme in C. gigas.     

Two epizootic Perkinsus spp. events in commercial oyster farms at Santa Catarina,Brazil

Perkinsus spp. have been detected in various bivalve species from north‐east Brazil. Santa Catarina is a South Brasil state with the highest national oyster production. Considering the pathogenicity of some Perkinsus spp., a study was carried out to survey perkinsosis in two oyster species cultured in this State, the mangrove oyster Crassostrea gasar and the Pacific oyster Crassostrea gigas. Sampling involved eight sites along the state coast, and oyster sampling was collected during the period between January 2013 and December 2014. For the detection of Perkinsus, Ray's fluid thioglycollate medium (RFTM) and histology were used, and for the identification of the species, PCR and DNA sequencing were used. Perkinsus spp. was found by RFTM in C. gigas and C. gasar from São Francisco do Sul. This pathology was also detected in C. gasar from Balneário Barra do Sul both, by RFTM and histology. Perkinsus marinus was identified in C. gigas and C. gasar from São Francisco do Sul and Perkinsus beihaiensis in C. gasar from Balneário Barra do Sul. This is the first report of P. marinus in C. gigas from South America. Results of this preliminary study suggest that both oyster species tolerate the species of Perkinsus identified, without suffering heavy lesions.     

Rearing equipment on the quality aspects and growth performance of Crassostrea gigas (Thunberg, 1793): Implications for marketability

Shell characteristics and consequent marketability of the oyster Crassostrea gigas are influenced by several factors, and most important of which are the rearing methods and gears. In this study, we investigated the effects of two different gears on the quality aspect (shell shape, weight meat percentage, backward shell shape, epibionts and blisters) and growth rate (weight and length) of C. gigas by adopting the innovative Ortac Oyster Farming System, during the field grow‐out phase of production. Aspect and growth of oysters farmed in Ortac and floating bags were compared. After 14 weeks, the aspect of the animals was influenced by the gear, resulting in a lower quality in the Ortac (i.e. worse shell shape, lower weight meat percentage, higher percentage of backwards and blisters) than in the floating bag, while the incidence of epibionts was less in the Ortac. Regarding the growth performance, total wet weight gained in the Ortac was lower (~28.3 g) than in the floating bag (~32.4 g; p < .05). These results highlight the efficiency of the floating bag as oyster rearing system in the Mediterranean basin and suggest the need to alter the Ortac gear to these environmental conditions.     

Comparative studies of the growth,survival, and reproduction of diploid and triploid Kumamoto oyster,Crassostrea sikamea

Triploid oysters have been used for farming to improve growth but have not been created in the Kumamoto oyster, Crassostrea sikamea, which is one of the crucial aquaculture species on the southern coast of China. In the present study, triploids were created using cytochalasin B to inhibit polar body II release in C. sikamea, with the untreated oysters as controls. Triploidy rates of 87 and 57.67%, on average, were obtained in larvae and adults, respectively. Larval growth and survival of the triploid were significantly lower than that of the controls (p < 0.05). In contrast, the triploid postlarvae and adults had a significant growth advantage over the controls (p < 0.05) during the period of 180 (December) to 450 days (September of the next year). Moreover, the triploids clearly exhibited significant sterility in the reproductive season. The glycogen and triglyceride contents in the gonad, adductor muscle, mantle, and gill were higher in triploids than in controls from 180 to 450 days. As a result, high physiological energy supply was strongly correlated with superior growth and reduced reproduction in triploid C. sikamea. The triploid C. sikamea is an excellent oyster species and can be used to improve growth for C. sikamea farming.     

Insights on the association between somatic aneuploidy and ostreid herpesvirus 1 detection in the oysters Crassostrea gigas,C. angulata and their F1 hybrids

Cytogenetic abnormalities associated with viral infections, including from viruses of the Herpesvirales order, have been reported in vertebrate species. Ostreid herpesvirus 1 (OsHV‐1) has been detected worldwide during mortality outbreaks of the Pacific oyster Crassostrea gigas. On the other hand, a high proportion of aneuploid cells in somatic tissues have been observed in C. gigas. In this study, we analysed the putative association between aneuploidy levels and the detection of OsHV‐1 in gills of C. gigas, the Portuguese oyster C. angulata and their F1 hybrids cultured in Ria Formosa (Portugal). OsHV‐1 was detected by PCR in 5.4% of the total of oysters analysed (n = 111) namely in 11.1%, 8.0% and 1.7% of C. gigas, C. angulata and F1 hybrid respectively. Sequencing analysis of a viral fragment amplified with the C2/C6 primer pair revealed a high similarity with the OsHV‐1 reference type. Moreover, in situ hybridization confirmed the presence of OsHV‐1 in gill tissue. Oysters where OsHV‐1 was detected had a significantly higher mean percentage of aneuploid cells (25%) than the ones where the virus was not detected (18%). However, the overall low percentage of positive samples contrasted with the high mean percentage of aneuploidy observed, with 50% of the oysters analysed showing a percentage of aneuploid cells between 20% and 30%. We hypothesize that somatic aneuploidy may adversely affect oysters making them more prone to OsHV‐1 infection, but the virus is unlikely to be the cause of somatic aneuploidy.     

A comparative study of the survival,growth and gonad development of the diploid and triploid Hong Kong oyster,Crassostrea hongkongensis (Lam & Morton 2003)

The Hong Kong oyster, Crassostrea hongkongensis is an economically important and widely cultivated oyster species in southern China. Aiming to improve the aquaculture potentials of C. hongkongensis, a triploid organism was generated by inhibiting the second polar body formation in the fertilized eggs, using 0.5 mg L^?1 cytochalasin B. Triploids exhibited significantly higher growth and were larger than the diploids throughout the life cycle. Much of the triploid growth advantage can be attributed to resource reallocation to structural development due to gonadal sterility. Most of the triploid oysters exhibited polymorphic sterility with atrophic gonads and abnormal gametogenesis. Only few ‘dysfunctional’ sperms were produced by the triploid males, thus ruling out reproductive and genetic contamination of natural oyster populations from triploid cultivations. This advantage in addition to the superior growth of triploids makes it a potent alternative in Hong Kong oyster aquaculture industry. Triploid survival was mostly found to be similar to that of diploids except for day 360, when a significant triploid advantage in terms of survival was noted. Furthermore, we found that growth and survival of Hong Kong oyster is significantly higher at the Zhuhai site in comparison to the Beihai site. Our results indicate a strong environmental effect in Hong Kong oyster cultivation and points to the fact that Zhuhai site was more suitable for oyster farming than the Beihai site. Overall, this study provides valuable information to the aquaculture industry of southern China.     

Broodstock conditioning of the Portuguese oyster (Crassostrea angulata,Lamarck, 1819): influence of different diets

The Portuguese oyster Crassostrea angulata shows great potential in oyster farming. The conservation of pure populations of this species is important for production diversification and biodiversity preservation. In this way, the zootechnological development for seed hatchery production is extremely important. Broodstock conditioning is a key step in the process of rearing bivalves in a hatchery. Many factors regulate the reproductive cycle, being food one of the most important ones. To evaluate the effect of different diets on C. angulata reproductive performance, broodstock were conditioned with different food regimes formulated fundamentally by flagellates (Diet 1 – Pavlova lutheri and Isochrysis galbana clone T‐ISO; Diet 2 – P. lutheri, T‐ISO and Skeletonema costatum) and constituted fundamentally by diatoms (Diet 3 – S. costatum and Chaetoceros calcitrans; Diet 4 – P. lutheri, S. costatum and C. calcitrans). During conditioning, samples of oysters were collected to evaluate condition index, gonadal development and biochemical composition. At the end of the conditioning period, oysters were induced to spawn to evaluate reproductive output (fecundity, fertilization rate and D‐larvae development). The diets had an impact on the gametogenesis process, energy storage and reproductive output performance, being the best results those obtained in broodstock fed with the diatoms‐predominant diets. However, those fed with diets majority flagellates had an unsuccessful performance. Holistic approaches incorporating all results in this study reveal and reinforce the idea that the diatom species used presented the nutritional requirements to C. angulata broodstock, being essential in the conditioning phase.     

Nutrient removal efficiency of Hydropuntia cornea in an integrated closed recirculation system with pink shrimp Farfantepenaeus brasiliensis

In this study, we have tested the effect of seaweed stocking density in an experimental seaweed biofilter using the economically important red seaweed Hydropuntia cornea integrated with the cultivation of the pink shrimp Farfantepenaeus brasiliensis. Nutrient removal efficiency was evaluated in relation to seaweed stocking density (2.5, 4, 6 and 8 g fw L^?1). Total ammonia nitrogen (TAN) was the main nitrogen source excreted by F. brasiliensis, with concentrations ranging from 41.6 to 65 μM of NH₄⁺‐N. H. cornea specific growth rates ranged from 0.8 ± 0.2 to 1.4 ± 0.5% day^?1 with lowest growth rates at higher seaweed stocking density (8 g fw L^?1). Nutrient removal was positively correlated with the cultivation densities in the system. TAN removal efficiency increased from 61 to 88.5% with increasing seaweed stocking density. Changes in the chemical composition of the seaweed were analysed and correlated with nutrient enrichment from shrimp effluent. The red seaweed H. cornea can be cultured and used to remove nutrients from shrimp effluents in an integrated multi‐trophic aquaculture system applied to a closed recirculation system. Recirculation through seaweed biofilters in land‐based intensive aquaculture farms can also be a tool to increase recirculation practices and establish full recirculation aquaculture systems (RAS) with all their known associated benefits.     

The obvious heterosis and genetic characters of intergeneric cross and backcross juveniles between blunt snout bream (Megalobrama amblycephala) and topmouth culter (Culter alburnus)

In this study, we used artificial insemination to generate hybrid groups of fish [MC‐F₁(MA♀×CA♂) and MC‐F₂(MC‐F₁♀×♂)] by intergeneric crosses of Megalobrama amblycephala (MA) and Culter alburnus (CA); sequential backcrosses [CAM‐B₁ (CA♀×MC‐F₁♂) and MCC‐B₁ (MC‐F₁♀×CA♂)] were also performed. All these hybrids showed high rates of fertilization, hatching and survival (p > 0.05). For genetic traits, compared with those of the M. amblycephala and C. alburnus parental lines (Table 1), the fertilization rate, hatching rate and 7‐day survival rate of MC‐F1(MA♀×CA♂), MC‐F2(MC‐F1♀×♂), CAM‐B1 (CA♀×MC‐F1♂) and MCC‐B1 (MC‐F1♀×CA♂) by artificial insemination exhibited similar high rates (p > 0.05). The morphology of the four hybrids MC‐F₁/F_2, CAM‐B₁ and MCC‐B₁ were intermediate between those of their parents. Compared with their parents of MA and CA, weight gain rate (WG), specific growth rate (SGR) and protein efficiency ratio (PER) of hybrids MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ were significantly (p < 0.05) increased and feed conversion ratio (FCR) was significantly (p < 0.05) decreased after 3 months feeding. Moreover, protein content of muscle for MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ was significantly (p < 0.05) higher and carbohydrate content of muscle was significantly (p < 0.05) lower than their parents. The females and males of the four hybrids had normal gonadal development. In this study, we successfully generated intergeneric and backcross hybridization lines with fertile potential among fish of the Cultrinae subfamily and these hybrids had obvious heterosis in terms of growth performance, feed utilization and muscle quality.     

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

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

Different responses between orange variant and cultured population of the Pacific oyster Crassostrea gigas at early life stage to temperature‐salinity combinations

Although breeding of rare shell colour variants has drawn widespread attention from shellfish breeders, the potential disadvantages of their adaptive capacity have been ignored in practice. To explore the difference in adaptive capacity between orange shell variant (OSO) and commercially cultured population (CPO) of the Pacific oyster Crassostrea gigas at early life stage, the development to D‐larvae and larval survival and growth (just 23 and 30°C for larval experiment) of them were compared under different temperature (16, 23 and 30°C) and salinity (17, 25 and 33 psu) combinations. In this study, at 23°C and 25 psu, for both OSO and CPO there was no difference in fertilization rates and survival (p > .05) (mean percentages of D‐larvae after fertilized 40 hr ≥ 95.00%; mean larval survival rates on day 10 > 80.00%). However, the percentage of D‐larvae of CPO at 40 hr was significantly (p < .05) higher than OSO at temperatures of 16 and 30°C and 25–33 psu and 17 psu at 23°C. Similarly, CPO has a better larval survival on day 10 and growth than OSO at salinities of 17 and 33 psu at 23°C. Overall, our results indicate that OSO can have an equally good performance like CPO at early life stage under optimal condition (23°C; 25 psu), but the potential disadvantages in adaptive capacity will be shown at suboptimal conditions. These findings can guide future hatchery breeding of OSO, and suggest the potential disadvantages in adaptive capacity in rare colour variants need more attention in further breeding.     

An artificial oyster‐shell reef for the culture and stock enhancement of sea cucumber,Apostichopus japonicus,in shallow seawater

Because of its high nutritional value and health benefits, aquaculture production of Apostichopus japonicus in China is the largest of any single species. Therefore, the development of new farming methods is of considerable significance. In this study, discarded oyster shells have been used to create an artificial reef for the culture of this species. The results have shown that from 6th March 2009 to 26th November 2009, the wet weight of sea cucumber increased from 49.57 ± 1.16 to 79.87 ± 1.46 g ind^?1. Between 16th July and 18th October, the specific growth rate and daily weight gain of A. japonicus differed significantly from other periods. Population density was higher within the reef compared with outside the reef area, and the difference was significant (P < 0.01). The maximum distance between A. japonicus individuals within the reef area on 3rd March and 16th July was 65.0 ± 3.3 and 62.9 ± 4.4 cm, respectively, but the difference was not significant (P > 0.05). The diatom species attached to the oyster shells were similar to those found in the stomach content of A. japonicus. In conclusion, the oyster‐shell reef provides a suitable habitat and shelter for the culture of the sea cucumber, A. japonicus.     

Technical and economic feasibility of integrating seahorse culture in shrimp/oyster farms

Integrated multi‐trophic aquaculture systems (IMTA) are building ecosystems designed to produce aquatic organisms with less environmental impact. We have developed a simple system and a management strategy for introducing seahorse (Hippocampus spp.) culture in shrimp/oyster farms to increase economic sustainability. The system includes broodstock, nursery and grow‐out phases of Hippocampus reidi G. culture. We performed a test in a 42.4‐ha marine shrimp/oyster farm, which showed that this system is technically feasible. Then, we performed a partial budget analysis including cost‐return, cash flow and financial feasibility analysis. More than 12 000 marketable size seahorses may be produced in 402 m² annually. Initial investment was only US$ 21 103.00. The benefit–cost ratio was ~US$ 20.00 for each US$ 1.00 spent, internal rate of return was 131% and payback period <2 years. In conclusion, the introduction of seahorse in shrimp/oyster IMTA system is technically feasible, profitable, resilient and shows high liquidity. This system may be easily established in developing countries and has potential to provide seahorses to supply different markets (live specimens for the marine aquarium trade and dried specimens for traditional Chinese medicine and the curio trade). In addition, it can indirectly contribute to preserve natural populations.