广西茅尾海的香港巨牡蛎(Crassostrea hongkongensis)在不同养殖区的性腺发育变化

为探究香港巨牡蛎(Crassostrea hongkongensis)的繁殖特性以及春季死亡原因,采用肉眼观察、活体镜检和组织切片方法研究了茅尾海沙井的香港巨牡蛎苗种吊养于大风江黄场的1龄牡蛎的性腺发育周年变化,并建立了快速判断香港巨牡蛎性腺发育程度的方法.对比分析了吊养在茅尾海沙井、大风江入海口内的黄场和入海口外的鑫丰场的2龄牡蛎性腺发育的启动和发展过程.结果显示,广西茅尾海的香港巨牡蛎每年只有1个繁殖周期,性腺发育过程可划分为未分化期(1-3月)、分化期(4月)、成熟排放期(5-10月)及休止期(11-12月)4个阶段;1龄和2龄牡蛎的性腺发育基本同步;大风江入海口外的牡蛎的性腺发育较入海口内的牡蛎提前约15d,而比茅尾海内海的牡蛎提前约30 d,入海口外的牡蛎的性腺4月即发育到成熟排放期,冬季饵料的丰富程度和温度、盐度等因素可能是造成这种差异的原因.研究还发现,高温少雨天气下,海水盐度高,抑制了牡蛎性细胞的排放,导致入海口外侧吊养牡蛎的大规模死亡,濒临死亡的牡蛎性腺处于退化和崩解状态.鉴于此,认为4月初即需密切监测吊养于入海口外的牡蛎性腺的发育状况和天气状况,及时收获性腺饱满的牡蛎,或将吊养牡蛎的蚝排放置在河流入海口内区域过冬.     

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.     

Application MALDI TOF on protein identification of vitellogenin in giant grouper (Epinephelus lanceolatus)

A new proteomics technology has been implemented to study the protein repertoires of developing oocytes of giant grouper (Epinephelus lanceolatus). Knowledge of the chemical composition and physiochemical properties of vitellogenin (Vtg) is necessary to interpret the functional and biological properties attributed during ovulation. Vtg, as a biomarker indicator in sex determination, has been analyzed to determine the sex and maturational status of fish in the absence of the gonad tissue. A male giant grouper was induced by 2 mg/kg of 17ß-estradiol (E2), and blood was sampled at days 0, 1, 3, 5, and 10. SDS-PAGE 1D electrophoresis was used to analyze Vtg protein, and Vtg identification was done with 4800 Plus MALDI TOF/TOF? mass spectrophotometer (Applied Biosystems/MDS SCIEX, USA). Meanwhile, MS/MS de novo sequencing identified the proteins by matching sequences of tryptic peptides to the known sequences of other species. Vtg was confirmed by MASCOT at 95 % significant level, and molecular mass was 187 kDa. Protein resolved on SDS-PAGE as a double band of approximately the same mass as determined with MALDI-TOF. The N-terminal sequences and identification of Vtg were also determined. The potential of using MS methods to understand the structure and function of Vtg is discussed.     

Gametogenetic cycle and reproductive effort assessed by two methods in 3 age classes of Pacific oysters, Crassostrea gigas, reared in Normandy

Two methods were used to estimate the reproductive output of female Pacific oysters reared in Normandy: histology with image analysis and ELISA (Enzyme-Linked ImmunoSorbent Assay) which allowed the quantification of egg protein. Condition indices, gonad area and gametogenetic stages of the oysters were determined in the entire population (males and females) between May and October 2005. All investigations were performed in 3 age classes: oysters in their first, second or third years (corresponding to spat, half-grown and market-sized oysters, respectively). Both quantitative histology and ELISA provided similar results in terms of reproductive effort (illustrated by the gonado-somatic index, GSI) except during the GSI drop, corresponding to spawning, which was less marked with the ELISA method. Growth depended on oyster age, the sex ratio was well balanced and the reproductive cycle was synchronized in all age classes. In the 3 age classes, most of the oysters were ripe and ready to spawn on August 8, and ten days after the post-spawning stage was observed in 40% of spat oysters and 70% of half-grown and market-sized bivalves. The major difference between age classes was observed in the reproductive investment, with spat having a lower reproductive output. For example, in males and females, the gonad area reached 78–79% in the median animal section at full maturity (August 8) in half-grown and marketable oysters while it attained only 59% in spat. At the same time, GSI in females was, respectively in spat and the 2 oldest age classes, 33% (quantitative histology)–36% (ELISA) and 55% (quantitative histology)–60% (ELISA). The mean assessed gonad weight and fecundities increased with the age of the oysters: 1.3 g and 12 million eggs, 7.8 g and 135 million eggs, and 11.5 g and 146 million eggs in spat, half-grown and market-sized oysters, respectively. Marked differences thus appear between 2 and 3-year-old oysters and spat. As early as their first reproductive cycle, the young oysters not only showed the reproductive features of the species in Normandy, but also a pronounced lower reproductive effort. This lower energy demand could explain their higher survival rate.     

Gametogenesis, reproductive investment, and spawning behavior of the Pacific giant oyster Crassostrea gigas: evidence of an environment-dependent strategy

The progress of gametogenesis was studied in oysters Crassostrea gigas having the same origin (Tremblade), but cultured during 1 year in two distinctive French marine areas, the Baie des Veys and Marennes-Oléron. We assessed seasonal changes in the reproduction cycle on the basis of stereological techniques to estimate reproductive investment and measurement of gonad evolution area by quantitative histology. From a qualitative point of view, both oyster groups presented typical reproductive stages, but showed differential timing, in particular during the sequence of spawning and duration of the re-absorption stage. Oysters in Baie des Veys had a single partial spawning in August and a re-absorption stage that extended until winter. Oysters in Merennes-Oléron had a partial spawning in July and massive release of gametes during August. Spawnings in both the areas were related to maximum temperature (19°C). The quantitative analysis showed, on an annual basis, a higher reproductive investment by oysters from Baie des Veys, 86% against 53% in the other group. Larger gonads, higher gamete production, and more intensive spawning were the characteristics of oysters in Baie des Veys. Recently, the reproduction pattern and investment has been related with summer mortalities; therefore, a quantitative understanding of reproductive processes becomes necessary for C. gigas. Environmental conditions at each site may explain differences in the progress and intensity of gametogenesis. While temperature regulated the time and speed of gametogenesis, results suggest that the intensity was influenced by the quantity of available food but may need further research. However, nutrient recycling from unreleased gametes in the gonads of oysters from Baie des Vey is a factor to be considered in the results of this study.     

Immunohistochemical observations of vitellin synthesis and accumulation processes in ovary of Ezo abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis>

Ovarian follicle cells are the site of yolk protein synthesis in the Ezo abalone Haliotis discus hannai. In this study, histological observations of ovarian follicle cells were conducted in H. discus hannai by immunocytochemical and in situ hybridization methods focusing on their function of yolk protein synthesis. An antibody raised against purified yolk protein (vitellin, Vn) recognized yolk granules in oocytes, and ovarian follicle cells adjacent to the oocytes under yolk accumulation were stained positively with both anti-Vn antibody and an antisense probe for vitellogenin (the precursor for vitellin, Vtg) mRNA. These results indicate that the abalone Vtg gene is transcribed and translated in the ovarian follicle cells. In oocytes in the early phase of yolk accumulation, positive reactions with the antibody appeared first in the stalk part, and the follicle cells adjacent to the stalk were also stained positively. These observations imply local transport of yolk protein from the follicle cells to the oocyte through an extracellular space around the oocyte stalk. Ovarian follicle cells changed their morphological characteristics and reactivity to the antibody in close relation to the stages of the adjacent oocyte, which suggests the presence of functional interactions between follicle cells and oocytes in the course of yolk protein synthesis and accumulation.     

Quantification of eggs and sperm in the Black-lip pearl oyster Pinctada margaritifera using an enzyme-linked immunosorbent assay (ELISA)

We have developed immunological probes to quantify eggs and sperm of the Black-lip pearl (BLP) oyster Pinctada margaritifera. The western blot assay revealed that the polyclonal antibodies developed in this study specifically recognized only egg and sperm proteins. These polyclonal antibodies also showed high sensitivities to the antigens, detecting 0.31–10 μg/ml of the egg or sperm proteins in an indirect enzyme-linked immunosorbent assay (ELISA). Accordingly, we used an indirect ELISA to quantify the eggs and sperm in BLP oysters collected in December 2009 from Weno Island in Micronesia and in May 2010 from Tahiti. The gonad somatic index (GSI), a ratio of gonad weight to somatic tissue weight, of the females collected from Weno Island ranged from 3.6 to 18.4 %, while the GSI of the females collected from Tahiti ranged from 5.6 to 12.6 %. Similarly, the GSI of the male BLP oysters from Weno Island ranged from 0.8 to 8.5 %, while the GSI of the males from Tahiti ranged from 4.8 to 7.5 %. These results lead to the conclusion that the immunological probes developed in this study can be successfully applied to quantify BLP oyster gonadal tissues and that these probes can be used in studies of BLP oyster reproductive ecology based on their sensitivity, rapidity, and affordability.     

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.     

Reproductive conditioning of Chilean scallop (Argopecten purpuratus) and the Pacific oyster (Crassostrea gigas): effects of enriched diets

The quality of diet fed to bivalve broodstock during reproductive conditioning directly affects growth and survival of their larval and postlarval progeny. The objective of the present study was to improve the quality of larval production in the Chilean scallop (Argopecten purpuratus) and the Pacific oyster (Crassostrea gigas) by testing of the effects of different experimentally enriched diets on the conditioning and early development of progeny under laboratory conditions. The diets used included protein-rich microalgae and mixtures of microalgae and lipid emulsions. Quality of D-larvae, pediveligers, and 1-mm postlarvae from the variously fed broodstock was measured to evaluate each diet as indicators of growth and survival of the early life stages. Biochemical analyses were carried out on eggs and larvae to determine the effects of the different diets on these parameters. The main results for the scallops and oysters were that a conditioning diet containing algae rich in protein improved larval growth and survival. This effect did not, however, extend past metamorphosis. Also, the best conditioning diet increased the lipid content of the eggs, as well as the protein content of the D-larvae, although the mechanism for this remains unclear.     

Age, growth and reproductive characteristics of round scad Decapterus maruadsi in the waters off west Kyushu, the east China Sea

ABSTRACT:   The purpose of this study was to determine the growth and reproductive characteristics of round scad Decapterus maruadsi in the East China Sea. The characteristics regarding growth and reproduction of round scad based on otolith reading and gonad histology, respectively, were estimated. The von Bertalanffy's growth model for round scad was estimated as follows:
FL _t  = 342[1 − exp{−0.55( t  + 0.58)}], (1 <  t  < 7)
where FL _t is fork length (mm) at age in years t . The maturation stage of the ovary was observed by histological techniques. June was the main spawning period; the maturation and migratory nucleus stage of the oocyte was observed when the gonad somatic index value was approximately greater than 4.     

Changes in fatty acid and sterol composition during oogenesis in the pearl oyster Pinctada margaritifera

The fatty acid and sterol composition of the oyster Pinctada margaritifera during oogenesis and in eggs was analysed. No major differences were observed during oogenesis, but the egg composition was significantly different from that of gonads. The amount of saturated fatty acids was the highest in eggs and the C16:0 predominant (P<5%); by contrast, the amount of 22:6(n‐3) was significantly lower (P<5%) than in gonads. No major differences were observed for the polar lipid (PL) composition during oogenesis. The main free sterols in gonads and eggs were cholesterol and brassicasterol. Among free sterols, the proportion of cholesterol diminished continuously from the beginning to the end of gonad maturation, and this decrease persisted in eggs after spawning. Cholesterol represented 40% to 55% of the sterol ester encountered in gonad and eggs. This study allowed us to investigate the fatty acid and sterol composition during oogenesis of the pearl oyster P. margaritifera, leading to a clearer understanding of the nutritional requirements of pearl oyster during the reproduction process.     

Quantitative measurement of reproductive output in the American oyster, Crassostrea virginica (Gmelin), using an enzyme-linked immunosorbent assay (ELISA)

Abstract. A quantitative gonadal index was developed for oysters, Crassostrea virginica (Gmelin. 1791), using polyclonal antibodies from eggs and sperm. Percoll used in the purification of oyster eggs and sperm greatly improved the purity of antigens compared to filtering the egg or sperm through a fine mesh only. The antigen-antibody reaction was tested with indirect sandwich ELISA using alkaline phosphatase-conjugated goat anti-rabbit igG as a secondary antibody. Rabbit anti-oyster egg IgG and anti-oyster sperm IgG initially exhibited a weak cross-reactivity over somatic tissue. Absoring with acetone-dried oyster tissue powder removed this cross-reactivity. Both antisera exhibited strong specific immunological reactions to oyster eggs or sperm respectively. The quantity of eggs or sperm was measured using ELISA and a quantitative gonadosomatic index (dry wt of egg or sperm/dry wt oyster) (GSI) calculated. GSI from ELISA correlated with gonadal stage measured histologically. Monthly mean GSI of female oysters was highest during late spring to early summer (0·157–0·201) and lowest during early winter to early spring (0·002-0·000). Maximum GSI observed during the study was 0·422 for female oysters and 0·446 for male oysters. Female oysters produce 3·7–65·4 million eggs, with an average of 21·1 million during each spawning. A positive correlation was observed between the number of eggs produced and oyster size; the number of eggs in the gonad increased as oyster size (i.e. total dry wt) increased (r= 0·67); however, the relationship was non-linear. Large oysters contained proportionally fewer eggs. Prevalence of Perkinsus marinus parasitism was high, 90–100%, during the study, as was weighted incidence, 1·33 to 2·67, No statistically significant correlation was observed between infection intensity and the per cent weight of oyster eggs or egg number.     

Characterization,fluctuation and tissue differences in nutrient content in the Pacific oyster (Crassostrea gigas) in Qingdao,northern China

The Pacific oyster (Crassostrea gigas) is one of the most important aquaculture species worldwide. Its meat quality is vital for consumer satisfaction, and nutrient content, especially glycogen, is closely associated with oyster fatness and meat colour. Fluctuations in nutrient content of short‐term starvation have not been previously reported, and seasonal variation in glycogen content in different tissues has rarely been reported. In the present study, we investigated these important aspects of oyster production and found that short‐term starvation (50 hr) did not significantly alter glycogen, protein or lipid content. The seasonal variation assay showed that glycogen and lipid accumulation was high in autumn and winter and that seawater temperature and protein content were inversely related to glycogen content. Glycogen content of the whole flesh was higher from January to April and was positively related to the condition index before the onset of gametogenesis. Glycogen content was higher in the gonad, labial palp and mantle compared to the gill or adductor muscle. Relative expression of genes encoding proteins involved in glycogen metabolism (glycogen synthase, glycogen phosphorylase, glycogen debranching enzyme and glycogen branching enzyme) was closely associated with the glycogen content in the corresponding tissues. Glycogen content in the gonad was regulated by glycogen metabolic and glycolysis pathway genes (6‐phosphofructo kinase, phosphoglycerate kinase, pyruvate kinase, hexokinase and glucose transporters), and stored glycogen was the main energy source for gametogenesis. These findings contribute to oyster aquaculture management and glycogen improvement and expand our understanding of glycogen metabolism in oysters.     

DNA methylation frequency and epigenetic variability of the Pacific oyster <Emphasis Type="Italic">Crassostrea gigas</Emphasis> in relation to the gametogenesis

The origin and maintenance of sexual gametogenesis might provide insights into eukaryote evolution. Epigenetic variation of eukaryote well-being would provide clues that could relate ecological and evolutionary processes. To investigate the relationship between gametogenic variability and epigenetic polymorphism in the Pacific oyster Crassostrea gigas, DNA methylation differences were represented by analyzing genome-wide methylation patterns using the fluorescent-labeled methylation sensitive-amplified polymorphism (F-MSAP) technique. The C. gigas genome was widely methylated and marked variations in the pattern of methylation were shown during female and male gonad development. Statistically, the total methylation level increased from early developing stage to resorbing stage, from 28.86 to 34.53% in ovaries, while it increased from 31.20 to 35.46% in this period in testes. The unweighted pair group method with arithmetic mean and principal coordinate analysis separated the C. gigas during female and male gonad development remarkably well, indicating remarkable differences across gametogenesis between ovaries and testes. Variation was higher in testes than in ovaries by the frequency based and multivariate analyses above. Both multivariate analyses and a significantly different population epigenetic structure suggested that a striking difference might exist across gametogenesis, reflecting a turning point of the methylation mechanism during reproductive progress. This study provides some information on the role of DNA methylation in the potential mechanism during female and male gonad development.     

Differentiation of gonad maturation in sibling precocious males of the sea trout (Salmo trutta m. trutta L.) in their first year of life

Age at maturity and dynamics of gonadal cycle vary among individuals in a standard population; the extent of that variability in a group of siblings is not known for any species. Therefore, the aim of the study was to find out how the differentiation of gonad development varies in a group of sibling fish. The study involved 176 seven-month-old sea trout parr (mean fork length of 11 cm) collected at a single time point during their first spawning season (late October) from the brook of Pniewa in Western Pomerania (Poland). Gonad maturation was determined from histological analysis which was interpreted according to specially developed criteria.

The stage and forms of gonad maturation were diverse in the sibling group studied. Gonads of 52.8% males were not maturing (Class I), with only spermatogonia A present. The remaining males (47.2%) had gonads differing widely in the development, from beginning of maturation to completion of the reproductive cycle (Classes from II to VIII). In addition to the classes described, some males were observed to feature attempted maturation. It is difficult to provide an unequivocal estimate of the proportion of precocious males (mature at a young age, prior to the first descent to the sea) in the first year of life because of difficulties in categorizing males in Class II. It was predicted that 32.4% of the population would complete the cycle during the first spawning season. The body length and weight of the maturing males was significantly smaller (P < 0.05) than that of the non-maturing males.

Three forms of gonad maturation were identified. Gonads of a large group of precocious males (70%) developed incompletely (only some germ cell developed during the reproductive cycle), some of those gonads assumed the form of attempted maturation and partial maturation. Testes of the remaining 30% of the males underwent complete maturation. Gonads that undergo attempted maturation do not spermiate.

It is only from microscope-aided analysis of the testes that the class of gonad development can be accurately evaluated and the extent of precocious maturation assessed, particularly in fish maturing for the first time. It is impossible to correctly identify maturing males of the underyearling sea trout on the basis of morphological examination of gonad appearance, abdominal massage to detect spermiation or gonadosomatic index value.     

长江口雌性成体刀鲚不同组织的脂肪酸组成分析

刀鲚(Coilia nasus)是一种江海生殖洄游性鱼类,长江口是刀鲚重要的洄游通道。本研究首次关注洄游至长江口卵巢发育比较成熟的繁殖群体的营养状况,并比较分析了其不同组织总脂和脂肪酸组成,为探明长江口水域刀鲚繁殖群体的营养状况及繁殖性能提供参考信息。结果显示,肌肉、肝脏和卵巢中的水分含量依次降低,总脂含量依次增加。饱和脂肪酸(SFA)总量在肝脏中最高,肌肉中最低,3种组织间差异显著(P<0.05);单不饱和脂肪酸(MUFA)中C18:1n9c的含量最高,其在肌肉、肝脏和卵巢中的含量依次为(40.88±0.19)%、(35.06±1.84)%和(42.85±2.14)%,肝脏中含量显著低于其他2个组织(P<0.05);多不饱和脂肪酸(PUFA)、n3-PUFA及二十二碳六烯酸(DHA)含量在肌肉、肝脏和卵巢中依次升高,卵巢中的含量显著高于肝脏和肌肉(P<0.05),而后二者间的差异不显著(P>0.05)。肌肉、肝脏和卵巢中不饱和脂肪酸(UFA)/SFA的比值依次为2.35、1.67和4.49,n3-PUFA/n6-PUFA的比值依次为4.94、3.87和5.13...     

Multiple vitellogenins and product yolk proteins in striped bass,Morone saxatilis: molecular characterization and processing during oocyte growth and maturation

The multiple vitellogenin (Vtg) system of striped bass, a perciform species spawning nearly neutrally buoyant eggs in freshwater, was investigated. Vitellogenin cDNA cloning, Western blotting of yolk proteins (YPs) using Vtg and YP type-specific antisera, and tandem mass spectrometry (MS/MS) of the YPs revealed the complex mechanisms of yolk formation and maturation in this species. It was discovered that striped bass possesses a tripartite Vtg system (VtgAa, VtgAb, and VtgC) in which all three forms of Vtg make a substantial contribution to the yolk. The production of Vtg-derived YPs is generally similar to that described for other perciforms. However, novel amino-terminal labeling of oocyte YPs prior to MS/MS identified multiple alternative sites for cleavage of these proteins from their parent Vtg, revealing a YP mixture far more complex than reported previously. This approach also revealed that the major YP product of each form of striped bass Vtg, lipovitellin heavy chain (LvH), undergoes limited degradation to smaller polypeptides during oocyte maturation, unlike the case in marine fishes spawning buoyant eggs in which LvHAa undergoes extensive proteolysis to osmotically active free amino acids. These differences likely reflect the lesser need for hydration of pelagic eggs spawned in freshwater. The detailed characterization of Vtgs and their proteolytic fate(s) during oocyte growth and maturation establishes striped bass as a freshwater model for investigating teleost multiple Vtg systems.     

Reproductive cycle and biochemical composition of the Zhe oyster Crassostrea plicatula Gmelin in an eastern coastal bay of China

The reproductive cycle and biochemical composition of the oyster Crassostrea plicatula in Rushan Bay, an eastern coastal bay of China, were investigated between March 2004 and February 2005 in relation to environmental factors (temperature, salinity and chlorophyll a). Histological analysis, combined with oocyte examination and measurements of protein, glycogen and lipid levels and RNA/DNA ratio from gonad, adductor muscle and mantle tissue of each sex were performed. The gametogenic cycle of C. plicatula comprised two phases: a resting phase (November-February) and reproductively active phase, including ripeness and spawning, during the rest of year. Gametogenesis in spring and summer was associated with an increase in protein and lipid contents in the ovaries, and took place at the expense of reserves (mainly glycogen) accumulated previously during winter. The RNA/DNA ratio is a good indicator of sexual maturity in the ovary; the increasing RNA/DNA ratio in the ovary appears to show the rising synthetic activity of vitellin within the ovary. Spawning occurred at the moment when water temperature and chlorophyll a levels were highest, and salinity lowest throughout the year. High variation in glycogen contents during storage and gametogenic development suggests that carbohydrates are the main respiratory substrate in this species. The results demonstrated that C. plicatula may be considered a conservative species in gametogenic pattern.     

Maturity assignment based on histology‐validated macroscopic criteria: Tackling the stock decline of the Mediterranean swordfish (Xiphias gladius)

1. Effective stock assessment relies on the evaluation of the reproductive potential of a population, but this is often achieved using fast, inexpensive, and inaccurate macroscopic indicators of gonadal maturity. Indeed, this approach suffers from high error rates as a result of misclassification and operator errors. Although histology is considered the gold standard to assign reproductive stages, it represents a time‐consuming and expensive method. Therefore, cost‐effective approaches are currently needed to assess the species‐specific reproductive potential and define the correct size at first maturity (L₅₀).
2. In the present study a histology‐calibrated method based on the gonadal index (GI) was developed to assign maturity stages to the overfished Mediterranean swordfish (Xiphias gladius), supported by histological classification of gonadal development, an estimation of L₅₀, and an analysis of the reproductive output of this species.
3. The performance of this new method had an overall agreement of 90% with histological data, and the L₅₀ calculated by the GI‐based method compared with histology analysis was not statistically different (p > 0.05). Moreover, in order to apply this method without bias, a thorough validation of the relationship between length and gonad weight was carried out and the suitability of the GI to compare individuals of different size and maturity stages was confirmed.
4. The present findings provide a comprehensive picture of the patterns characterizing Mediterranean swordfish reproduction and a promising method to assign maturity stages based on a macroscopic indicator. This method has the potential to be extensively applied during routine fish sampling activities in the field because of its simple requirements.