Farming triploid oysters

Although the commercial benefits of triploidy have been evaluated in the Pacific oyster Crassostrea gigas (Thunberg, 1793), eastern oyster C. virginica (Gmelin, 1791), Sydney rock oyster Saccostrea glomerata (Gould, 1850) and European flat oyster Ostrea edulis (Linnaeus, 1750), so far this technique has only been commercialised for Pacific oysters.

Commercial production of triploids on the West Coast of North America began in 1985. Since then production of triploids has greatly increased and the use of tetraploid males to fertilise eggs from diploids to produce batches of 100% triploids has been developed. In 1999/2000, triploid Pacific oysters made up 30% of all Pacific oysters farmed on the West Coast of North America. Some hatcheries now use tetraploid males instead of chemical or physical stress to produce triploids. The rapid uptake of triploid and tetraploidy techniques has been facilitated by the almost total dependence that these oyster industries have on hatcheries for the supply of seed. This industry in the Pacific Northwest of the US and in British Columbia, Canada, would not have developed to its current size without hatchery seed supplies. Triploids are preferred over diploids in summer because diploids are less marketable when in spawning condition.

There was only limited interest in triploidy production in France until 1999, when IFREMER began to make sperm from tetraploids available to commercial hatcheries. In 1999/2000, only 10% to 20% of all the hatchery-supplied Pacific oyster spat were triploids, but with the use of sperm from tetraploid oysters, this could increase sharply.

Elsewhere around the world, the commercial uptake of triploid oysters has been slow to develop. However, in countries where the production of Pacific oysters is based on hatchery supply of seed, it is likely that with the use of tetraploid oysters, the farming of triploid oysters will increase in the near future.     

Production of triploid eastern little tuna,Euthynnus affinis (Cantor, 1849)

Herein, we developed a triploidization method using spawned eggs collected immediately after spawning of eastern little tuna (ELT), Euthynnus affinis. ELT broodstock induced the spawning by hormonal treatment in the tank, with the resulting spawned eggs being used for the triploidy induction. Under optimal conditions, the mean ± SEM triploidization and hatching rates were 97.2 ± 2.8% and 84.5 ± 10.3%, respectively. Although triploid ELT showed growth performance equivalent to that of diploids, the triploids died at a higher rate than the diploids during 2–4 weeks post‐hatching when triploids and diploids were reared in the same tank. Therefore, we propose that it would be necessary in a practical operation to use triploid‐only ELT seedlings to avoid selective cannibalism by the diploids. The ELT triploids exhibited an all‐female phenotype. Because previous studies have reported that female triploids show a greater probability of sterility than male triploids, this characteristic could be a major advantage. Since this triploidization method, using spawned eggs, can be performed without handling the broodstock, it is possible to avoid the physical damage caused by the process of artificial insemination, making it possible to repeatedly produce triploid populations without valuable broodstock loss. Thus, we have developed an efficient method to produce ELT triploids, although further study is essential to evaluate sterility of the triploid ELT.     

不同育性三倍体长牡蛎性腺发育过程中的营养成分比较

为阐明不同育性三倍体长牡蛎性腺发育与营养成分变化的关系,实验对不育型和可育型三倍体长牡蛎性腺发育过程中的主要营养成分(糖原、总蛋白质和总脂肪含量)进行分析,并与二倍体长牡蛎进行比较.结果显示,三倍体长牡蛎性腺—内脏团、闭壳肌和外套膜3种组织中的糖原含量均显著高于同时期的二倍体长牡蛎,性腺—内脏团和闭壳肌中的总蛋白质含量...     

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.     

微卫星标记在长牡蛎三倍体鉴定中的应用

为开发一种简单有效的倍性检测方法对长牡蛎三倍体诱导结果进行准确评估,本实验采用细胞松弛素B抑制第二极体释放诱导产生长牡蛎三倍体,选用7个微卫星位点扩增基因组DNA,通过亲子代基因分型进行倍性检测,检测结果采用流式细胞仪加以验证,以评估微卫星标记倍性检测的准确性。另外,本研究探讨了准确鉴定倍性所需的微卫星标记数量与微卫星—着丝粒重组率(y)之间的关系,以期为其他物种采用分子标记进行倍性检测提供理论依据。结果显示,细胞松弛素B诱导产生的115个长牡蛎子代经7个微卫星位点鉴定得到40个三倍体,与流式细胞仪检测结果一致,准确率达到100%,7个微卫星位点的(1-y)的乘积为0.005。随机挑选6个位点,也可鉴定出所有三倍体,(1-y)的乘积为0.005~0.042。研究表明,本研究中开发的微卫星标记可以简单高效地鉴定长牡蛎三倍体,微卫星位点的(1-y)的乘积小于0.005,倍性检测的准确率可达100%,采用微卫星标记进行倍性检测对于加速长牡蛎三倍体育种进程具有十分重要的意义。     

Covariation between diploid and triploid progenies from common breeders in rainbow trout, Oncorhynchus mykiss (Walbaum)

The covariation between diploid and triploid progenies from common breeders was investigated in various progeny-testing experiments where either dams or sires were sampled from rainbow trout, Oncorhynchus mykiss (Walbaum), stocks. Triploidiza tion was found to frequently reduce the performance in the traits studied: body length and weight, growth, coefficient of condition and pyloric caeca number. Triploidization also generated significant interactions with the parental breeding value. These interactions were caused in part by the familial variance not being the same in triploids as in diploids, but also by actual ranking differences between diploid and triploid familial performances. However, the effect of these interactions was minor as compared with the amount of variation common to both ploidy levels (genetic correlations averaged 0.7–0.9). Therefore, selection of diploid breeders appeared efficient enough for improving triploid progeny, unless family selection methods including triploid progeny testing were preferred for other reasons. Lastly, it was observed that variances from maternal origin tended to be larger in triploids, whereas variances from paternal origin tended to be smaller, as compared with diploids. This point was discussed referring to the genetic make-up of triploids and in the scope of dams and sires for selective breeding.     

Aquaculture performance of triploid barfin flounder Verasper moseri

ABSTRACT:   To evaluate the aquaculture performance of triploid barfin flounder Verasper moseri , the sex ratio, maturation, growth and the relative proportion of body parts were examined. The sex ratio of triploids was similar to diploids under communal rearing conditions, but the proportion of female diploids was higher than that of triploids under separate rearing conditions. The gonadosomatic index of triploid females was very low even during the spawning season, and the ovaries were rudimentary. These results suggest that triploid barfin flounder females were sterile. In addition, triploid males produced a small quantity of milt containing very few spermatozoa with abnormal shapes. Spermatozoa obtained from triploids were aneuploidies. When normal eggs were fertilized with sperm from triploid males, no fry developed. These results suggest that triploid barfin flounder males were functionally sterile. Triploid males grew more slowly than diploid males, and triploid females showed similar or slower growth than diploid females, whether reared separately (23 months) or communally (35 months). The ratios of visceral weight to the edible parts for triploid males were similar to those for diploid males, but ratios for triploid females were higher than for diploid females during the spawning period. In conclusion, a significant improvement of growth was not found in triploid barfin flounders.     

Growth of triploid oyster, Crassostrea madrasensis (Preston)

The performance of I and II meiotic triploids and control oysters (Crassostrea madrasensis) reared at Tuticorin Bay was compared to determine if the improvements in the growth of edible oysters were additive to faster growth in triploids. After a grow‐out period of 12 months, both mean whole weights and shell heights were in order I meiotic triploid>II meiotic triploid>control. Mean whole weights and shell height of different oyster lines were all significantly different (P<0.05). On an average, larger morphological traits indicated that growth improvements from triploids were additive, and throughout the study triploid oysters maintained faster growth rate than their diploid siblings. Condition index and adductor muscle diameter of both triploids were higher than those of control.     

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.     

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.     

The Potential for Developing Controlled Breeding in the Chilean Oyster

International interest in the commercial aquaculture of the Chilean oyster, Tiostrea chilensis, is frustrated by an inability to artificially control their unusual breeding behaviour. The manipulation of breeding in this oyster will rely on a better understanding of its gametogenic cycle. Consequently, the gametogenesis of a wild population of highly fertile Chilean oysters from northern New Zealand was studied over three years to assess the potential for artificially controlling oyster breeding for aquaculture. The population was mostly comprised of simultaneous hermaphrodite oysters that generally followed annual cycles of gametogenesis. Oysters spawned as males throughout each year with increased spawning from November to February (ie., austral spring and summer). Ova were abundant throughout the year, although female spawning increased during spring. The unpredictable spawning of ova among individual oysters and the strong possibility of self-fertilization create difficulties for the development of controlled breeding in this species. If the full aquaculture potential of T. chilensis is to be realized, further research should be directed at identifying the factors involved in controlling female maturation, spawning and fertilization.     

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.     

Effect of triploidy on paternal and maternal variance components in brown trout, Salmo trutta L.

The covariation between diploid and triploid progenies from common breeders and the effect of triploidy on the parental variances were investigated in brown trout (Salmo trutta L.) using two progeny testing experiments, sampling, sires and dams respectively, from the same population. The traits studied were body weight, growth, condition factor and red spotting of the skin. Triploidization generated some interactions with the parental breeding value, but their effect was minor (less than 20% of the genetic variance, in most cases) as compared with the amount of variation common to both ploidy levels. These interactions were mainly caused by a scale effect, triploidy reducing the variance attributable to sires and increasing the variance attributable to dams. Actual lack of correlation (genetic correlation significantly less than 1) between diploid and triploid familial performances was observed in a single instance out of 18. The modification of respective parental variance components by triploidy, already observed in the rainbow trout Oncorhynchus mykiss (Walbaum), appears as a logical consequence of the genetic make‐up of triploids, and should be taken into account in selective breeding.     

Microsatellites markers to depict the reproductive and genetic patterns of farmed gilthead seabream (Sparus aurata): illustration by a case study on mass spawning

In the absence of breeding strategy, natural spawning constitutes the breeding ground for fish farmers to empirically manage their commercial broodstock. In this context, we used six microsatellite markers to characterize the genetic pattern of six commercial seabream broodfish tanks having a common history spanning four generations. The progeny of one tank single‐day mass‐spawning event, reared in two different environments, was used to estimate the genetic parameters for body weight. Limited genetic differentiation was observed among broodfish groups. A panel of nine loci allowed us to unambiguously assign 95.4% of the offspring (1692) and identify 37 parents (65% of the total broodfish). The limited effective population size (N_e = 15.3) was due to the elevated variance of parental contributions and to broodfish failing to contribute to the progeny. The fluctuation of the allele frequency highlighted the risks of genetic drift and reduction in the heterozygosity in the next generations. Heritability for body weight was moderate at commercial size (0.40 ± 0.10) and the high genetic correlation at later stages laid the groundwork for precocious selection criteria for growth. The discussion opens on the opportunity to use mass spawning for selective breeding.     

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

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

熊本牡蛎中国群体与美国群体杂交效应及杂交三倍体优势分析

为了评估熊本牡蛎(Crassostreasikamea)中国群体与美国群体的杂交效应与杂交三倍体优势,构建了杂交群体和自交群体,并使用细胞松弛素B诱导了杂交三倍体,比较分析了幼虫、稚贝与成贝的生长与存活优势。自交群体和杂交群体的幼虫、稚贝与成贝的培养条件均相同,室内培育幼虫,培育密度为4～5个/mL,自然海区养殖稚贝与成贝,养殖密度为40～45个/吊。结果表明,与自交组相比,杂交二倍体具有较高的卵裂率(13.61%)与D幼率(5.67%)(P0.05),但杂交二倍体幼虫的壳高生长表现杂种劣势(–0.43%),而稚贝、成贝的壳长与壳高的生长表现杂种优势,平均优势率分别为3.96%与6.65%。杂交三倍体诱导组幼虫的壳高生长优势率为3.69%,稚贝及成贝的壳长与壳高平均生长优势率分别为12.69%与13.64%,并且日龄越大生长优势越显著。总体上,杂交三倍体诱导组在3～180日龄具有存活劣势,并且15日龄存活劣势率最大(–48.72%),而在360日龄存活优势率为6.70%。杂交二倍体幼虫与成贝均具有存活优势,平均优势率分别为10.44%与4.59%。本研究表明熊本牡蛎中美地理群体杂交二倍体的生长和存活优于自交二倍体,而杂交三倍体诱导组的生长优势较显著,并且在成贝期具有显著的成活率优势,表明杂交三倍体诱导组的优势来源于三倍体优势和部分杂种优势。     

Effect of Timing of Seed Deployment on Growth and Mortality of Oysters, Crassostrea virginica, Affected by Juvenile Oyster Disease (JOD)

Abstract.— Mortality of oysters, Crassostrea virginica , caused by Juvenile Oyster Disease (JOD) has been a chronic problem the northeastern United States since 1988. To assist in identifying potential health management options, the effect of time of spawning and deployment of oysters on resultant growth and mortality was examined in the Damariscotta River, Maine. Broodstock oysters from a line selected for fast growth were spawned at 2-wk intervals between 28 March and 19 July 1995. Duplicate groups of juvenile oysters from each spawn were deployed in floating trays between 23 May and 31 August. Mortality and size of all cohorts were determined from late June to early December. Cumulative mortality of oysters was related to the timing of deployment. Cohorts placed in the river before June or after mid-August had cumulative mortalities of 20% or less. Oysters deployed between 8 June and 18 August had cumulative mortalities ranging from 64–96%. Growth of all cohorts was rapid immediately after deployment but decreased once JOD began to have an effect. Final size of oysters was directly related to age. This study determined that seed oysters placed in the Damariscotta River between June and mid-August will experience high mortality and low growth. Potential management options to reduce JOD impact include 1) early spawning and deployment in May, ensuring that a mean shell height >25 mm is reached prior to August and 2) late spawning and deployment after mid-August to avoid exposure to JOD.     

The breeding potential and biochemical composition of triploid Manila clams, Tapes philippinarum Adams and Reeve

In 1989 and 1990. triploid Manila clam, Tapes philippinarum Adams and Reeve, seed were reared to 15-20 mm at the Fisheries Laboratory, Conwy, and planted out in the Menai Strait, North Wales. In each of the summers of 1992 and 1993, three of these batches, at 2, 3 or 4 years old, were returned to the laboratory to assess ploidy, size, spawning potential and biochemical composition. Percentage triploidy at this time was similar to that in the seed. After 6 and 8 weeks of warmwater conditioning. Only 45 out of l21 triploids (37%) were induced to spawn by thermal shock, with only one spawning as a male. By comparison, 75% of diploid clams spawned with a 1:1 ratio of males to females. Mean fecundity of triploids was significantly lower than that for diploids, at 0.497 compared with 1.54 million eggs per female. Compared with eggs from diploid females, eggs from triploids were larger and significantly fewer of them developed into D-larvae when fertilized by sperm from diploid males. Triploid clams were heavier and had a higher condition index and carbohydrate content than diploids of the same age, but lipid levels were similar. Potential advantages of producing and cultivating 100% triploid batches of Manila clam seed are discussed.     

Reproductive performance and mature gonad morphology of triploid and diploid Black Tiger shrimp (Penaeus monodon) siblings

Sibling harvest age Black Tiger shrimp triploids and diploids of both sexes were reared to reproductive maturity, crossed with wild caught females and males, conditioned for spawning and a comprehensive reproductive performance trial was undertaken. Ovarian development, spawning frequency, fecundity, hatch rate, gonad morphology, male reproductive tracts and thelycum impregnation rates of the wild female × triploid male cross were assessed. After ablation, ovarian development and cycling between wild G₀ diploid and G₁ diploids was not significantly different, whereas G₁ triploids failed to show any signs of ovarian development and cycling, thus resulting in no G₁ triploid female spawnings. There were 10 G₀ diploid female × G₀ diploid male first‐spawnings and 9 G₀ diploid female × G₁ diploid male first‐spawnings, all of which produced viable nauplii. In comparison, there were 7 G₀ diploid female × G₁ triploid male first‐spawnings, none of which produced viable nauplii. The 26 wild G₀ diploid female spawnings had more eggs than the 1 G₁ diploid female spawning. Gonad morphology and male reproductive tract assessments showed impaired reproductive development in triploid gonadal tissues of both sexes (compared with sibling diploids and wild shrimp) to a point where complete maturation had not occurred. The thelycum of 16 wild G₀ diploid females crossed with G₁ triploid males had no visible spermatophore present, suggesting that G₁ triploid males are incapable of developing viable spermatophores and mating with females. This study demonstrates that the triploid females and males are incapable of producing viable gametes and are thus reproductively sterile.     

Studies on aneuploid pearl oyster (Pinctada martensii Dunker) produced by crossing triploid females and a diploid male following the inhibition of PB1

Viable aneuploid embryos of pearl oyster, Pinctada martensii Dunker, were produced by inhibiting the first polar body (PB1) with cytochalasin B treatment in eggs from triploids fertilized with haploid sperm. During the period of growth measurement, aneuploid showed the slowest growth compared with diploid and triploid groups. The body size and weight measurement data showed that there were no differences between aneuploids (as a group) and diploids in body size and weight (P>0.10), but that they were significantly different from triploids (P<0.01). There were no differences between aneuploids within diploid condition (2n±n) and diploids in SL (P>0.1), but significant differences in BW were found (P<0.05). Aneuploids within triploid condition (3n±n) were significantly smaller than triploids in BW (P<0.05), but not different from triploids in SL (P>0.05), and almost identical to diploids in both (P>0.1). These dada indicated that some aneuploids might be associated with growth retardation. Karyotype analysis revealed that there were metacentric, submetacentric or subtelocentric chromosomes losses or gains; aneuploid pearl oysters with the same chromosome numbers had different chromosome composition. Aneuploids are valuable research materials for genetic analysis.