Comparative biochemical composition in gonad and adductor muscle of triploid and diploid catarina scallop (Argopecten ventricosus Sowerby II, 1842)

Biochemical components of gonad and adductor muscle for diploid and triploid catarina scallop, Argopecten ventricosus, were evaluated and compared at four periods in 1 year (January, April, June, and October). Two comparisons were done. The first one compared an untreated control (diploid) vs. a triploidy-treated group for which the percentage of triploids was 57%. The second comparison was done on a group derived from within the triploidy-treated group, separating diploids (internal control) from triploids ('putative triploids'). Regardless of which comparison, in the gonad diploid scallops had larger concentrations of proteins, carbohydrates, lipids, and acylglycerols than triploid scallops. This reflects the maturation processes in diploid scallops vs. the sterility seen in most triploid scallops, and it is particularly supported by the consistently larger concentration of acylglycerols in gonads of diploids than in triploids. The gonad index of the internal control (diploid) group was significantly larger than that seen in the putative triploid group at all sampling periods but October, when none of the gonad biochemical components were different between ploidy groups.Triploid scallops had a significantly larger muscle index than diploids from April to October. This can be caused by a larger gain in muscle tissue in triploids than diploids from January to June. However, there were no consistent differences in any of the biochemical components evaluated in adductor muscle of diploids and triploids. The use of freshly ingested food rather than reserve mobilization from muscle in diploids is suggested by these results. Nutrients derived from ingested food are apparently used for muscle growth in triploids, whereas in diploids those nutrients serve primarily for gonad development. The importance of freshly ingested food for maintenance and growth is suggested because the decline in biochemical components seen in October in both muscle and gonad was paired with a decline in weight of those two organs, especially when the control groups are considered, but a decrease was also evident for the triploid groups. This may have been caused by the presence of El Ni?o, with its characteristic high water temperatures and low productivity.     

二倍体和三倍体太平洋牡蛎鳃扫描电镜的比较

利用扫描电子显微镜技术对二倍体和三倍体太平洋牡蛎(Crassostrea gigas)鳃的表面结构进行了观察和比较。结果显示：三倍体牡蛎鳃丝的宽度、鳃丝间的距离较二倍体大;鳃丝的微细结构比二倍体更致密;鳃丝间通过丝问连接形成的孔洞大于二倍体。这些不同表明二倍体和三倍体呼吸及摄食可能存在差异。     

Haematological parameters in Umbrina cirrosa (Teleostei, Sciaenidae): a comparison between diploid and triploid specimens

Haematological features were compared between diploid and triploid specimens of the ray-finned fish Umbrina cirrosa. No significant differences between diploids and triploids were reported in haematocrit and total haemoglobin concentration, but erythrocytes and thrombocytes were significantly greater in size in triploids. Glycaemia was significantly lower in diploids, whereas triploid erythrocytes were more resistant to osmotic stress. In triploids, a greater fraction of leukocytes was positive for alkaline phosphatase activity, when stimulated with Bacillus clausii spores, otherwise no significant increase of oxygen consumption was observed in triploid leukocytes after stimulation, based on assays for superoxide anions. Triploids were characterized by a lower concentration of circulating blood cells with a lower surface/volume ratio when compared with diploids. These features may lead to a general disadvantage of triploids in withstanding stress conditions: a situation that needs to be taken into account in aquaculture practice.     

黄姑鱼三倍体的诱导、鉴定及其性腺发育特征

研究采用冷休克方法诱导黄姑鱼(Nibea albiflora)三倍体并进行鉴定,进一步采用组织学和生殖相关基因表达分析比较了三倍体黄姑鱼的性腺发育特征。研究结果表明:(1)在受精后2.5min以3℃进行10min的冷休克处理,冷休克处理组的受精率和孵化率分别为(70.31±4.49)%和(21.5±6.63)%,其受精率和孵化率显著低于二倍体对照组;(2)经流式细胞仪倍性检测和染色体核型分析发现,三倍体的DNA含量为二倍体的1.5倍,染色体数目为72条,而二倍体的染色体数目为48条,三倍体的比例为100%;(3)三倍体的生殖腺指数显著低于二倍体,进一步通过组织切片观察发现三倍体的精巢和卵巢发育较二倍体滞后,在12月龄时,二倍体精巢和卵巢处于Ⅴ期,而三倍体精巢和卵巢分别处于Ⅲ期和Ⅰ期;(4)三倍体精巢的dmrt1和vasa基因及三倍体卵巢的cyp19a基因表达量均显著低于二倍体(P<0.05);三倍体卵巢的vasa基因表达量也比二倍体低(P>0.05)。综上结果表明:研究通过冷休克处理成功诱导了黄姑鱼三倍体;三倍体的性腺发育较二倍体滞后,育性明显降低。     

Sexual maturation in triploid rainbow trout, Salmo gairdneri Richardson

This paper compares some morphological and endocrinological characteristics of diploid and triploid rainbow trout.
Significant differences were found between diploid and triploid females in GSI, condition factor, gut weight, liver weight and percentage dress-out, and between diploid and triploid males in GSI, condition factor and gut weight.
Diploid females had large, well-developed ovaries containing yolk-filled secondary oocytes whereas the triploids had only string-like ovaries containing nests of oogonia. No primary oocytes were present.
All the diploid males produced copious quantities of milt but it was possible to express a thin, watery milt containing motile spermatozoa from only two of the 12 triploid males. Testes weights in triploids were similar to those of diploids but, while the diploid testes were packed with spermatozoa, those of the triploids consisted mainly of spermatocytes and spermatids with few spermatozoa present. Measurements of the heads of spermatozoa revealed that those from triploids were larger and had a wider size range than those from diploids.
Levels of testosterone and 11-ketotestosterone in triploid and diploid males were not significantly different. However, levels of testosterone and 17β-oestradiol in diploid females were considerably higher than those of triploid females.     

Size-Structures and Reproductive Characteristics in Diploid and Triploid Populations of Disporum sessile D. Don (Liliaceae)

Abstract Disporum sessile (Liliaceae), a perennial herb of temperate forests is composed of diploid (2n=16) and triploid (2n=24) populations. The size structure differed remarkably as triploid populations had few small plants and no seedlings. Triploid flowering plants were considerably larger than diploids. Triploids that flowered were 2.5 times larger than diploids that flowered and the size of vegetative ramets produced by triploids was twice as large. In triploids, fruiting rates were quite low only with inviable seeds and vegetative propagule size was greater than that of diploids. As regards growth parameters that help to increase plant size, triploids were superior to diploids. Differences in growth and reproductive parameters between diploids and triploids may contribute to forming different patch sizes.     

A population dynamic model for facultative agamosperms

Plants that can reproduce both sexually and agamically are called facultative apomicts. Some species, such as Taraxacum, contain both sexual diploids and triploid facultative apomicts. Triploids produce seeds without gamete fusion and recombination, and can also produce pollen and fertilize diploids. We present a population dynamic model that deals with gene flow and competition between diploids and triploids, with differing allocation towards reproductive investment in seeds and pollen. This paper examines whether diploids and triploids of plants with facultative agamospermy can coexist within a single population. We analyse the global behavior of such a dynamic system. Features of the system are significantly affected by the germination rates of diploids and triploids. Either diploids or triploids persist alone when the germination rate of diploids is sufficiently larger or smaller than that of triploids, respectively. Competitive exclusion occurs when both germination rates are sufficiently large. Coexistence is possible under certain specific conditions when: (I) the germination rates of both diploid sexuals and triploids are not sufficiently large, and (II) triploids produce sufficient pollen. When diploid sexuals and triploids coexist, triploids cannot exist alone, implying that the pollen of triploids is necessary to exploit diploid ovules.     

The haematology of triploid landlocked Atlantic salmon, Salmo solar L.

Triploid landlocked Atlantic salmon had a larger mean erythrocyte volume but lower erythrocyte count than diploids; the haematocrit was the same in diploids and triploids. Although the total blood haemoglobin content and the mean corpuscular haemoglobin concentration were lower in triploids than in diploids, the actual mean corpuscular haemoglobin content of triploid erythrocytes was higher than that of diploids. The increase in triploid mean erythrocyte volume was mainly due to an increase in cell length; there was only a minor increase in cell width and no increase in cell height. The nucleus of triploid erythrocytes occupied a greater percentage of the corpuscular volume than did the diploid nucleus. Mean cytoplasmic haemoglobin concentration was found to be the same for diploids and triploids when this was taken into account     

Morphological comparisons and hypotheses on the origin of polyploids in parthenogenetic Fasciola sp

It is known that Fasciola sp. from Japan and the Republic of Korea consist of diploids (2n = 2x = 20), triploids (2n = 3x = 30), and mixoploids with diploid and triploid cells. Triploids are distributed over Asia and Hawaii. Abnormal spermatogenesis and parthenogenetic reproduction are the main characteristics of Fasciola sp. Here we measured 21 different morphological parameters of diploid and triploid flukes of Fasciola sp. obtained from Japan and the Republic of Korea. Statistical analysis showed that diploid and triploid flukes were morphologically different. No bivalents or trivalents could be detected in diploid and triploid flukes, respectively. Based on our findings, we speculate that parthenogenetic diploids, triploids, and mixoploids (2x/3x) of Fasciola sp. are genetically related to each other.     

A hypothesis on the origin of genetic heterozygosity in diploids and triploids in Japanese Cayratia japonica species complex (Vitaceae)

We previously reported the occurrence of triploid strains in Japanese populations of Cayratia japonica (Thunb.) Gagnep. Interestingly, the triploid and most diploid strains had variably reduced pollen fertility. Two questions emerged from this earlier work: (1) How do triploids arise, and are they allotriploids or autotriploids? and (2) Why is there low pollen fertility in some diploid plants? We used a molecular genetic approach to determine the phylogenetic origins of triploids in C. japonica and the closely related species Cayratia tenuifolia (Wight & Arn.) Gagnep. In our analysis, we compared the sequences of the nuclear single-copy genes LEAFY and ASYMMETRIC LEAVES1. As a result, most triploids and diploids were heterozygous for the loci examined; the triploid genome shared an allele with the diploid genome, but other alleles differed between the ploidies. Therefore, Japanese populations of C. japonica and C. tenuifolia almost certainly arose from repeated hybridization events among genetically differentiated strains. Using our sequence data, we discuss possible scenarios accounting for the occurrence of triploids in the two species of Cayratia.     

Muscle fibre growth dynamics in diploid and triploid rainbow trout

The effect of triploidy on muscle fibre growth was determined by comparing hyperplasia and hypertrophy of white muscle fibres in all-female, diploid and triploid rainbow trout Oncorhynchus mykiss (100–400 mm total length). Conventional morphometry and protein and DNA concentrations were used to assess muscle fibre hyperplasia and hypertrophy in white muscle samples derived from an anterio-dorsal location. Muscle fibre distributions were significantly different between triploids and diploids in trout <300 mm. The proportion of fibres <20 μm was higher in diploids than in triploids and the proportion of fibres in the 20–40 μm category was higher in triploids than in diploids. This indicates that the hyperplastic fibres of triploids are larger than those of diploids. Larger hyperplastic fibres in triploids are probably due to the combined effect of increased nuclear size in triploids and the relatively high nucleus: cell ratio observed in small muscle fibres. These larger fibres may be less favourable to cellular metabolic exchange because of their smaller surface area to volume ratios, and perhaps account for reduced viability and growth observed in triploids during early life stages. On the other hand, the lack of difference in the distribution of fibres <20 μm between diploids and triploids at larger body size ranges (301–400 mm) imply that triploid trout may have higher rates of new fibre recruitment and growth capacity at these sizes. There was no difference between diploid and triploid trout in the mean size of muscle fibres; however, the number of fibres per unit area was reduced by 10% in triploids. No differences were observed in protein or DNA concentrations in muscle tissues between the two genetic groups. Since triploid nuclei have 1·5 times more DNA than diploid nuclei, this deviation from the expected muscle DNA concentration (1·3–1·4 times more DNA in triploids when the 10% reduction in fibre density is considered) suggests that the number of nuclei per muscle fibre is reduced. In both diploids and triploids, mean fibre size increased with body length while fibre density decreased. Similarly, protein concentration in the muscle tissue increased and DNA concentration declined with increasing body length. Protein/DNA ratio was strongly and positively correlated with fibre size. These results demonstrate that changes in DNA and protein concentrations can be used to assess hyperplasia and hypertrophy in muscle tissues. However, the morphometric procedure provides better insight into muscle fibre growth as it enables the direct visualization and analysis of muscle fibre distribution patterns.     

Sex Determination and Polyploid Gigantism in the Dwarf Surfclam (Mulinia Lateralis Say)

Mulinia lateralis, the dwarf surfclam, is a suitable model for bivalve genetics because it is hardy and has a short generation time. In this study, gynogenetic and triploid. M. lateralis were successfully induced. For gynogenesis, eggs were fertilized with sperm irradiated with ultraviolet light and subsequently treated with cytochalasin B to block the release of the second polar body (PB2). Triploidy was induced by blocking PB2 in normally fertilized eggs. The survival of gynogenetic diploids was very low, only 0.7% to 8 days post-fertilization (PF), compared with 15.2% in the triploid groups and 27.5% in the normal diploid control. Larvae in all groups metamorphosed at 8-10 days PF, and there was no significant post-larval mortality. At sexual maturation (2-3 months PF), all gynogenetic diploids were female, and there was no significant difference (P > 0.05) in sex ratio between diploids and triploids. These results suggested that the dwarf surfclam may have an XX-female, XY-male sex determination with Y-domination. Compared with diploids, triploids had a relative fecundity of 59% for females and 80% for males. Eggs produced by triploid females were 53% larger (P < 0.001) in volume than those from diploid females. In both length and weight measurements at three months PF, the gynogenetic diploids were not significantly (P > 0.33) different from normal diploid females, suggesting that inbreeding depression was minimal in meiosis II gynogens. Triploid clams were significantly larger (P < 0.001) than normal diploids. We hypothesize that the increased body-size in triploids was caused by a polyploid gigantism due to the increased cell volume and a lack of cell-number compensation.     

Why are triploid zebrafish all male?

Adult triploid zebrafish Danio rerio has previously been reported to be all male. This phenomenon has only been reported in one other gonochoristic fish species, the rosy bitterling Rhodeus ocellatus, despite the fact that triploidy is induced in numerous species. To investigate the mechanism responsible, we first produced triploid zebrafish and observed gonad development. Histological sections of juvenile triploid gonads showed that primary growth oocytes were able to develop in the juvenile ovary, but no cortical alveolus or more advanced oocytes were found. All adult triploids examined were male (n = 160). Male triploids were able to induce oviposition by diploid females during natural spawning trials, but fertilization rates were low (1.0 ± 3.1%) compared with diploid male siblings (67.4 ± 16.6%). The embryos produced by triploid sires were aneuploid with a mean ploidy of 2.4 ± 0.1n, demonstrating that triploid males produce aneuploid spermatozoa. After confirming that adult triploids are all male, we produced an additional batch of triploid zebrafish and exposed them (and a group of diploid siblings) to 100 ng/L estradiol (E2) from 5 to 28 dpf. The E2 treated triploids and nontreated triploids were all male. The nontreated diploids were also all male, but the E2 treated diploids were 89% female. This demonstrates that triploidy acts downstream of estrogen synthesis in the sex differentiation pathway to induce male development. Based on this and the observations of juvenile gonad development in triploids, we suggest that triploidy inhibits development of oocytes past the primary growth stage, and this causes female to male sex reversal.     

Effect of triploid fitness on the coexistence of diploids and tetraploids

The conditions for the coexistence of diploids, triploids and tetraploids in a single population were investigated with a deterministic model under the assumptions that diploids might produce 2 n gametes, and that triploids had a lower fitness than other cytotypes and generated equal proportions of haploid and diploid gametes. When diploids produced only haploid gametes, the dynamics of the cytotypes were similar to that of heterozygote disadvantage with two alleles at a single locus, with triploids being equivalent to the heterozygotes. Production of 2 n gametes by diploids increased the pool of diploid gametes and created a stable equilibrium involving a majority of diploids and a minority of polyploids. When the fitness of tetraploids was equal to or higher than that of diploids, increased triploid fitness decreased the threshold of 2 n gametes necessary to deterministically fix tetraploids in the population. Conversely, when tetraploids were less fit than diploids, the rate of 2 n gamete production leading to the exclusion of diploids first decreases and then increased with increasing triploid fitness. Triploids are repeatedly found in diploid-tetraploid hybridizations and are rarely totally sterile. They might play a determinant role in the future of multiple cytotype populations. The effect of triploids depends on the relative fitness of diploids and tetraploids and is also a function of their fitness.     

Influence of environmental parameters on growth pattern and population structure of Carassius auratus gibelio in Eastern Ukraine

Population structure and growth parameters of Prussian carp (Carassius auratus gibelio) were studied in 12 freshwater ecosystems of the Donbass region (Eastern Ukraine). These ecosystems differed significantly with respect to their surface area, water transparency and annual concentrations of phosphorus and nitrogen. Amongst the studied ecosystems, diploid males and females as well as a smaller percentage of triploid females were found. The population structure of C. a. gibelio varied significantly in terms of the percentage of triploids and sex ratio amongst diploids. A considerable proportion of triploid females (>10%) was found in four ecosystems with intermediate surface area (38–50 ha) and relatively high growth rate of specimens. The sex ratio amongst diploids was significantly female-biased in seven of 12 ecosystems, including those where triploids were present in considerable numbers. The growth parameters of Prussian carp were significantly correlated with ecosystem characteristics, but the growth patterns of diploids and triploids were not significantly different from each other. The restricted distribution of triploid specimens suggests that the triploid form might counter diverse environmental challenges, whereas the diploid form of Prussian carp seems to be efficiently adapted to a wide range of ecosystem conditions.     

Comparison of methods for hatchery-scale triploidization of European catfish (Silurus glanis L.)

Heat-, cold- and hydrostatic pressure shocks were applied in order to improve triploidy induction in European catfish ( Silurus glanis L . ). A 41°C heat shock (45 s, starting 9 min after gamete activation) provided 88% triploids and a high percentage of malformation (38.8 ± 4.1%). The superior 6°C cold shock (20 min, starting 9 min after gamete activation) gave 100% triploids and a 33.4 ± 3.8% triploid yield. The earliest hydrostatic treatments (600 kg cm²), lasting 4 min and starting 3 min after gamete activation, gave 97.8 ± 1.8% triploids and a 33.7 ± 16.9% triploid yield. The ploidy level was investigated using four approaches: karyotyping, quantification of Ag-stained nucleoli per cell, flow cytometry, and erythrocyte nuclear sizing by computer-assisted image analysis. Induction of triploidy under mass conditions in three experiments gave triploid percentages of 74%, 83% and 66%. Five months later, the percentage of triploids significantly decreased to 12.4%, 8.2% and 21.4%. The growth performance of yearlings was better in diploids than in triploids. Differences between diploids and triploids were 13.5% (NS), 27.6% (P   < 0.001) and 25.4% (P   < 0.05) in the three experiments. Analysis of variance showed the influence of ploidy (P   < 0.001) on growth rate, and multiple range analysis (LSD) assessed differences between total diploids (12.6 g) and total triploids (9.5 g) at the P   < 0.01 level.     

Immunohistochemical localization of IGF-I,IGF-II and MSTN proteins during development of triploid sea bass (Dicentrarchus labrax)

The cellular localization of IGF-I, IGF-II and MSTN proteins was investigated during ontogenesis of triploid sea bass (Dicentrarchus labrax) by an immunohistochemical approach. The results were compared with those observed in diploids. IGF-I immunostaining was mainly observed in skin, skeletal muscle, intestine and gills of both diploids and triploids. From day 30 of larval life, IGF-I immunoreactivity observed in skeletal muscle, intestine, gills and kidney was stronger in triploids than in diploids. At day 30, triploids exhibited a standard length significantly higher than the one of diploids. Although IGF-II and MSTN immunoreactivity was detectable in different tissues and organs, no differences between diploids and triploids were observed. The spatial localization of IGF-I, IGF-II and MSTN proteins detected in this study is in agreement with previous findings on the distribution of these proteins in diploid larvae and fry. The highest IGF-I immunoreactivity observed in triploids suggests a possible involvement of ploidy in their growth performance.Key words: IGF-I, IGF-II, MSTN, immunohistochemistry, triploid.     

Comparative growth performance of diploid and triploid European sea bass over the first four spawning seasons

During their 3–4 first years of life, triploid sea bass Dicentrarchus labrax grew in a similar fashion to diploids in fork length but more slowly than diploids ( P <0·05) in body weight, even when the diploids reached full sexual maturity. However, from 48–53 months of age triploids exhibited non-significantly higher instantaneous growth rates, and thus when fish were 4 years or older, differences in weight with diploids were no longer apparent, suggesting that triploidy could be of benefit in the culture of large (>1 kg) sea bass. The condition factor was reduced in both ploidies during the spawning season which took place in winter when the temperature was low. These observations suggest that any growth advantage in triploids, which were functionally sterile, may be offset by unfavourable environmental conditions. Thus, the potential gain of triploid fish, because they do not direct energy to gonadal growth, could not overcome the effects of low temperature on somatic growth, which coincided with the spawning season. This suggests that the low growth of this species during winter is more a consequence of low temperature than of the energetic cost associated with reproduction. On the other hand, the lower hepatosomatic index in triploid females in contrast to diploid females might be indicative of the lack of gonadal oestradiol-mediated hepatic synthesis of vitellogenin. Also, erythrocyte and haematocrit measurements showed an increased nuclear and cellular volume in triploids, but with similar cell numbers to those of diploids, respectively.     

The effect of triploidy and vaccination on neutrophils and B-cells in the peripheral blood and head kidney of 0+ and 1+ Atlantic salmon (Salmo salar L.) post-smolts

Sterile triploid fish are being used in aquaculture to prevent early unwanted sexual maturation and the genetic interaction between wild and cultured fish; however, triploid fish are typically considered to be more susceptible to disease than diploid counterparts. Proportions of leucocytes from the head kidney and peripheral blood were identified using monoclonal antibodies and flow cytometry in triploid and diploid, vaccinated and unvaccinated, out-of-season (0+) and 1+ Atlantic salmon (Salmo salar L.) three weeks post seawater transfer. Triploid 1+ fish were significantly (P<0.05) heavier than diploid fish at the time of sampling, whereas triploid 0+ had a significantly lower condition factor than diploids. Ploidy had a significant effect on the proportion of B-cells in the blood of both 0+ and 1+ fish, and the head kidney of 1+ fish, with triploids having lower proportions of B-cells to diploids in both smolt groups. In addition, a significant ploidy×vaccination interaction effect was observed in the response of neutrophils in the blood (vaccinated diploids had a higher mean proportion than diploid unvaccinated) and B-cells in the head kidney (in vaccinated fish, triploids had a lower mean proportion than diploids) in 0+ smolts. Vaccination was found to significantly increase the proportion of B-cells in the head kidney of 1+ smolts in both ploidy. Size (fish weight) was positively correlated with neutrophil proportions in 1+ fish. Our findings are discussed in relation to the physiological differences related to ploidy. The results suggest that ploidy as well as smelting regime influences the immune system of Atlantic salmon post-smolts.     

Shock-induced triploidy and its effect on growth and gonad development of the European catfish, Silurus glanis L.

Triloidy was induced in European catfish, Silurus glanis L., by cold-shocking eggs at 4°C for 30 and 40 min respectively, starting 5 min after fertilization. The hatching success of cold-shocked eggs was 25–30%. Cold shocks longer than 1 h caused total mortality. The triploid character of the cold-shocked European catfish was proved by karyological and red blood cell size analyses. The gonads of the triploid fish were significantly smaller than those of the diploids, while the growth rate values of the triploids were significantly higher than those of the diploids.