Life history of a simultaneously hermaphroditic fish,Diplectrum formosum

Sand perch Diplectrum formosum were collected in the Atlantic Ocean waters off the south‐eastern U.S. for life history analyses between April 2001 and July 2004 and ranged in size from 63 to 236 mm standard length (L_S) and 0–8 years of age. Diplectrum formosum are simultaneous hermaphrodites that reach 50% sexual maturity at 12·2 months and 122·8 mm L_S for testicular tissue and 13·6 months and 129·3 mm L_S for ovarian tissue. The gonad contains ovarian and testicular tissue separated by a thin basement membrane, with no means of internal self‐fertilization. Spawning females were found between March and January with a peak of spawning activity in May and a realized spawning periodicity of 2 days, equivalent to a maximum of 168 spawning events per year. Fish with testicular tissue in spawning condition were obtained throughout the year in every month sampled with a maximum frequency between June and September. There were also trends indicating that testicular tissue was more likely to be in spawning condition in the presence of hydrated oocytes within the accessory structure. This may indicate the use of the accessory structure as a storage site for hydrated oocytes until a mate can be located.     

Annual changes in fine structure of inner epithelial lining of the ovary of a marine sculpin,Alcichthys alcicornis (Teleostei: Scorpaeniformes), with internal gametic association

Alcichthys alciocornis has a viscous ovarian fluid in the ovarian cavity, which plays an important role in its unique mode of reproduction called internal gametic association (i.e., internal insemination and sperm-egg association but a delay in the physiological fertilization until spawning). Seasonal changes in fine structure of the inner epithelial lining and capillary endothelium of the ovary revealed that ovarian fluid originated as a result of the secretory activity of the tissues. The ovarian cavity of A. alcicornis is lined with an ovigerous lamella epithelium and an ovarian wall epithelium. During the spawning period, both epithelia actively secreted proteinaceous substances which seemed to constitute the ovarian fluid. The substances appear to be synthesized in the rough-surfaced endoplasmic reticulum from the material which was transported from the blood capillary, taken into the epithelial cells by endocytosis, accumulated in secretory vesicles via Golgi apparatus in the cells, and finally released into the ovarian cavity by exocytosis. Microapocrine secretion was also observed to occur in both epithelia. Secretory activity of both epithelia by exocytosis and microapocrine secretion showed distinct seasonal changes. Active exocytosis and microapocrine secretion were observed during the spawning period (April–May). These activities slightly declined during the degeneration period (May–June) and were lost during the early recovery period (July). During the mid to late recovery period (October–March), there was some exocytosis but no microapocrine secretion. © 1995 Wiley-Liss, Inc.     

Reproduction of the wedge sole (<Emphasis Type="Italic">Dicologoglossa cuneata</Emphasis> Moreau) in captivity: spawning parameters and influence of the natural temperature

In this work, the wedge sole (Dicologoglossa cuneata) spawning under a natural thermocycle has been studied. The spawning parameters were similar to those of other flatfish in captivity, the relative fecundity being very high (1.7 ± 0.1 × 10⁶ eggs kg female⁻¹ per spawning season). The percentage of floating eggs decreased with time and was directly related to egg diameter, daily relative fecundity and fertilization rate. Nevertheless, the hatching rate variations were not correlated to any other spawning parameter. The temperature range during the spawning season was very wide (11–21°C) though the largest spawns occurred between 12 and 16°C. The natural temperature cycle influenced wedge sole egg production, the winter temperature minimum indicating the onset of spawning, but the variations in the reproductive season did not appear to affect the daily egg production. Future research should focus on the photoperiod and its interaction with temperature as key factor for spawning.     

Effects of different artificial motile activating media on sperms motility of Waigieu seaperch Psammoperca waigiensis throughout a reproductive season

We previously determined changes in sperm quality of Psammoperca waigiensis during its spawning season and the optimal cation concentrations and osmolality for sperm preservation of this species at the peak of the reproductive season. In this study, we went one important step further by assessing the effects of the most adequate medium, considering the dilution ratio, osmolality, and cations (Na⁺, K⁺, Mg²⁺, and Ca²⁺) on the motility of P. wasigiensis sperm collected during the early, peak, and late spawning season. We determined the maximum velocity (VAP), and percentage of sperm motility (MOT), and the duration of sperm motility (DSM). Under optimal dilution, temperature, pH and osmolarity, MOT, VAP, and DSM did not statistically differ during early, peak, and late spawning season. However, under suboptimal external conditions, MOT, VAP, and DSM showed inconsistent trends during different spawning periods. We recommend using one of three different artificial motile activating media: (1) 0.55 M Na⁺, (2) 0.6 M K⁺ or (3) 1200 mOsm/kg for early; or (1) 0.6 M Na⁺, (2) 0.6 M K⁺ or (3) 1100 mOsm/kg for the peak; and (1) 0.65 M Na⁺, (2) 0.55 M K⁺ or (3) 1200 mOsm/kg for late spawning season; all at the dilution of 1:150 (v:v of semen: artificial motile activating medium).     

Fertilization environment of the non-copulating marine sculpin, Hemilepidotus gilberti

To clarify the extracellular environment for external fertilization in the non-copulating marine sculpin Hemilepidotus gilberti, sperm motility was measured in NaCl, KCl, mannitol solutions, seawater, and ovarian fluid. Spermatozoa of H. gilberti actively moved in seminal plasma the moment they were removed from the genital papilla. Spermatozoa showed higher motility in NaCl solution at osmolalities between 300–400 mOsmol kg^-1. In KCl and in mannitol solutions, spermatozoa actively moved at osmolalities between 500 and 800 mOsmol kg^-1, and at osmolality 300 mOsmol kg ^-1, respectively. The ovarian fluid was a transparent and viscous gelatinous material, rich in sodium with an osmolality of 340 mOsmol kg^-1. Sperm motility in the ovarian fluid lasted more than 90 min, which was six times longer than in seawater. This sperm motility under conditions isotonic to body fluid is similar to that of copulating marine sculpins rather than to other non-copulating marine fishes. In addition, eggs of H. gilberti could be fertilized in the ovarian fluid. This suggests that external fertilization takes place under physiological conditions similar to the internal conditions of the ovary provided by the ovarian fluid, which isolates the eggs from sea water for several hours after spawning. This manner of fertilization is thought to be one of the evolutionary pre-adaptations allowing copulation among marine sculpins.     

盐度渐变与骤变对脊尾白虾渗透、代谢及免疫相关酶活力的影响

为探讨盐度变化对脊尾白虾(Exopalaemon carinicauda)渗透、代谢及免疫相关酶活力的影响,实验设置了盐度渐变和骤变两个实验。渐变实验,设置5、10、15、20、25、30、33(CK)、40和45共9个盐度梯度;骤变实验,盐度从33突变至0、5、15、25和45,检测血清ATP酶(包括Na+/K+-ATP酶和总ATP酶)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)及超氧化物歧化酶(SOD)活力。结果表明,渐变情况下,盐度为5时,ATP酶活力出现最高值,然后随着盐度的升高表现出先降低后升高的趋势。总ATP酶活力在盐度为15—30之间较稳定,并在此范围内达到最低值。AKP和ACP活力几乎不受盐度渐变的影响。SOD活力随盐度的升高,先上升后下降,并在盐度为33时达到最大值。骤变情况下,ATP酶活力随时间波动较大,AKP和SOD随时间波动较小,而ACP几乎不受影响。结果说明,盐度骤变对脊尾白虾酶活力的影响较盐度渐变明显,ATPase和SOD活力更易随盐度而变化,代谢酶(AKP、ACP)受盐度变化的影响较小,说明渗透调节和免疫相关酶活力对盐度变化反应敏感,养殖过程中应尽量保持盐度稳定。     

Seminal plasma features of Prochilodus lineatus and Brycon orbignyanus throughout two consecutives spawning seasons

This study aims to describe seminal plasma characteristics, detect changes during and between two consecutive spawning seasons (SS), and compare plasma features between two important South American fish species. Prochilodus lineatus and Brycon orbignyanus sperm was collected over two (SS1; SS2). Each season was divided into first and second sampling periods (P1; P2). Thus, the four experimental periods were referred to as SS1P1, SS1P2, SS2P1, and SS2P2. Seminal plasma was analyzed for osmolality, pH, and Na⁺, K⁺, and Ca²⁺ concentration. Additionally, sperm concentration, motility rate, and velocities (curvilinear = VCL; straight line = VSL) were determined and correlated with plasma features. In P. lineatus, plasma osmolality was lower in SS1P2, pH was higher in SS2P2, Na⁺ was higher and K⁺ and Ca²⁺ were lower in SS2P1 compared with other experimental periods. Positive correlations were observed between motility and plasma osmolality, motility and Na⁺, and VCL and Na⁺. In B. orbignyanus, plasma osmolality was higher in SS2P1 and SS2P2 and K⁺ concentration was higher in SS1P1 compared with other experimental periods; no correlation was observed. Seminal plasma parameters change during SS; therefore, the composition of a sperm extender and artificial fertilization methods should be adapted to maximize fertilization rates.     

Variation of diet and food consumption of the scaldfish Arnoglossus laterna(Walbaum, 1792)

Although Arnoglossus laterna (Walbaum, 1792) is a common benthic fish in Portugal, several aspects of its feeding ecology remain incomplete. In this study, diet was examined and the food consumption estimated on inshore waters in the central coast of Portugal. The diet of the scaldfish included mainly mysids, amphipods and polychaetes, while the decapod Philocheras bispinosus Hailstone was the most frequent prey. Variation in the diet according to fish size (two size‐classes), sex and spawning/non‐spawning seasons was examined. Significant differences in diet composition were found between seasons, whereby the most frequently consumed prey in the non‐spawning season were crustaceans, while polychaetes and nemerteneans comprised most of its diet in the spawning season. The average value of the vacuity index was very similar between seasons (24.7% non‐spawning season and 25.0% spawning season). Diet overlap between size‐classes and sexes was high, but low between the two seasons. Food consumption of A. laterna was estimated based on diet characterization and evacuation rate. The estimate of the daily food consumption was 1.15 mg dry weight for the non‐spawning season and 1.67 mg dry weight for the spawning season. Significant differences in consumption rates were found between size‐classes and sexes. Larger individuals showed a higher consumption rate than smaller individuals. Females consumed more food than males in the non‐spawning season, while in the spawning season males consumed more than females.     

Role ofS gene product of bacteriophage lambda in host cell lysis

Studies with the induced lysogens of λS ⁺R⁺, λS^-R⁺, λS⁺R^- and λS-R^- phages have shown that while theS gene product is essential for the action of intracellularR gene product to release the periplasmic alkaline phosphatase in the presence of EDTA, the latter gene product can bring about this effect while acting onEscherichia coli cells from outside, in the absence of functionalS gene product; chloroform, could help the intracellularR gene product in effecting bacterial lysis in the absence ofS gene product. These result support the premise that theS gene product facilitates theR gene product in crossing the cytoplasmic membrane into the periplasmic space such that the latter can act on the peptidoglycan layer of the host cell thus causing both the release of alkaline phosphatase and cell lysis. An erratum to this article is available at .     

Response of sex ratio to timing of breeding in the small cyprinid Gnathopogon caerulescens

The influence of hatching date on the sex ratio of wild Gnathopogon caerulescens was examined. Cohorts reared from eggs collected in the early and middle parts of the spawning season showed almost balanced sex ratios, with female bias in some cohorts. Cohorts born later in the season mostly displayed male bias, and the mean proportion of males later in the season was significantly higher than in early‐ and mid‐season cohorts. These results indicate that the sex ratio of G. caerulescens changes with the time of breeding, increasing along with the ambient water temperature of the lake.     

Role of Zinc in Rhizobacteria-Mediated Suppression of Root-Infecting Fungi and Root-Knot Nematode

Understanding the environmental factors that influence the rhizosphere and inner root colonization of the disease‐suppressive strains of fluorescent pseudomonads is an essential step towards improving the level and reliability of their biocontrol activity. Soil amendment with Zn at 0.8 or 1.6 mg/kg of soil alone or in combination with Pseudomonas aeruginosa IE‐6S⁺significantly reduced nematode penetration in tomato roots. Zn applied alone did not reduce root infection caused by Macrophomina phaseolina or Fusarium solani but did reduce when used in combination with IE‐6S⁺. Soil amendment with Zn at 0.8 or 1.6 mg/kg of soil alone or in conjunction with IE‐6S⁺ markedly suppressed Rhizoctonia solani infection. Plant height, fresh weight of shoot and protein contents of the leaves substantially improved when used with Zn, however, plants growing in the soil treated with 1.6 mg/kg of Zn in the absence of IE‐6S⁺ not only reduced plant growth but also showed necrotic symptoms on the leaves. Zn application in the soil decreased populations of IE‐6S⁺ both in the rhizosphere and root. A positive correlation between bacterial rhizosphere and inner root colonization was also observed. With an increase in nematode densities in the soil, nematode penetration and subsequent galling due to Meloidogyne javanica increased. Regardless of the nematode densities, Zn applied alone or in combination with IE‐6S⁺ caused marked suppression of M. javanica. At all the population densities of M. javanica, Zn enhanced the efficacy of IE‐6S⁺ to reduce nematode invasion and subsequent gall development. IE‐6S⁺ caused significant suppression of soil‐borne root‐infecting fungi both in Zn‐sufficient and Zn‐deficient soil although this suppressive effect accentuated in Zn‐sufficient soils. In the absence of IE‐6S⁺ and/or Zn, increased nematode densities in the soil significantly reduced plant height, fresh weight of shoot and protein contents of the shoots. With an increase in nematode densities, populations of IE‐6S⁺ in the rhizosphere and root increased regardless of the Zn application. However, Zn‐deficient soils supported larger populations of IE‐6S⁺ compared with those of Zn‐sufficient soils.     

Immune responses and gene expression in hepatopancreas from Macrobrachium rosenbergii challenged by a novel pathogen spiroplasma MR-1008

Freshwater prawn Macrobrachium rosenbergii inoculated with 100 μl novel pathogen spiroplasma strain MR-1008 in logarithmic phase (10⁸ spiroplasmas ml^?1) were examined for alkaline phosphatase (AKP) activity, acid phosphatase (ACP) activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, as well as expressions of 7 immune related genes in hepatopancreas after 1–28 d. Hematoxylin-eosin (HE) staining showed obvious pathological features in hepatopancreas connective and epithelial tissue. Enzyme activity analyze showed that hepatopancreas AKP and ACP activity increased markedly (P < 0.05) when inoculated with spiroplasma MR-1008 after 5 d and 10 d, respectively. SOD enzyme activity changed less obviously and slightly increased at 1 day post-inoculation, but CAT activity decreased significantly after 5 d inoculation. The expression levels of lipopolysaccharide and β-1,3-glucan-binding protein (LGBP), peroxinectin (PE), α2-macroglobulin (α2M), AKP, ACP, CAT, and copper/zinc SOD (Cu, Zn-SOD) genes in the hepatopancreas were examined by Real-Time PCR (qRT-PCR) and the results demonstrated that these immune related genes were induced by challenge with spiroplasma MR-1008. The results suggested that the prawn immune responses could be activated or inhibited by spiroplasma MR-1008, and that the hepatopancreas also plays key roles in innate immunity for defense against the pathogen.     

Various methods to determine the gonadal development and spawning season of the purplish Washington clam, Saxidomus purpuratus (Sowerby)

The monthly variation in reproductive cycle, condition factor (fatness), gonad index (GI), ovarian egg diameter and biochemical composition [RNA, DNA content and their ratio RNA/DNA (R/D)] were compared to determine the spawning season of the purplish Washington clam, Saxidomus purpuratus, collected from the west sea of Taean, Korea. All these factors were correlated to the spawning season from May to October. Monthly variation and concentration of the R/D ratio especially represented indicative information about the processes of gonad development and spawning patterns. The increases in R/D ratio in the ovaries corresponded with increases in ovarian egg diameters and the GI during the spawning season. The peak of R/D and RNA content in females was a good indicator of sexual maturation and the DNA content in males. During winter between November and January, the value of fatness, GI, egg size, RNA and DNA content were lower than in other seasons, indicating depletion of energy reserves and lower metabolic demands because of colder temperatures.     

Genetic basis of tristyly in tetraploid Oxalis alpina (Oxalidaceae)

The inheritance of style‐morphs was investigated in tetraploid populations of tristylous Oxalis alpina (Oxalidaceae) to determine if alleles controlling style‐morphs are expressed at duplicated loci. In tetraploid populations, a dominant S allele leads to expression of the short‐styled phenotype at the short/non‐short locus and is epistatic to the M allele at the mid/long locus. The M allele results in expression of the mid‐styled phenotype but only if the S allele is absent. Long‐styled morphs are homozygous recessive at the short and mid loci. Test crosses of many tetraploid short‐styled individuals resulted in segregations of short‐, mid‐ and long‐styled individuals which, because of linkage between the short and mid loci, can only occur with polyploidy and expression of alleles at duplicated loci. Segregation patterns from three crosses suggest the possibility of disomic inheritance via preferential pairing of chromosomes in tetraploid populations of O. alpina. Segregation patterns in the progeny of mid‐styled individuals indicated that only a few individuals had more than one copy of the M allele, despite the potential for accumulation of M alleles via self‐fertilization of partially self‐compatible mid‐styled morphs in some populations. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 308–318.     

Physiological Performance of Asymptomatic and Yellow Leaf Syndrome-affected Sugarcanes in Venezuela

Serological analyses revealed the presence of the sugarcane yellow leaf virus (ScYLV) in asymptomatic (S^–) and symptomatic (S⁺) yellow leaf syndrome‐affected sugarcane plants of the cultivars PR.692176, C.323–68, V.64–10, V.71–47, V.75–6, SP.72–2086, SP.72–1210, SP.74–2005, C.323–68, B.80–549 and B.82–363. Tests for the presence of the sugarcane yellows phytoplasma, carried out by Dr P. Jones (IACR‐Rothamsted), gave negative results in all cultivars. Physiological analyses were performed in the top visible dewlap (TVD) leaf of S^– and S⁺ plants of the cultivar PR.692176. All plants were at the second ratoon and flowering. When compared with S^– plants, the S⁺ plants showed: (a) a marked reduction in the area of the leaf and internodes; (b) a high accumulation of total reducing sugars (TRS), glucans and α‐amino‐N in the leaf blade and of TRS in the corresponding leaf sheath; (c) a decrease in the chlorophyll, phosphorus and nitrogen content in the leaf; (d) the disappearance of the leaf diurnal fluctuations in TRS accumulation and export as well as the daily oscillations of TRS and glucans between dawn and dusk; and (e) major ultrastructural alterations in the companion cells of the phloem, including the accumulation of ScYLV particles in the cytoplasm. In S^– plants, none of the growth and physiological alterations described above were observed, in spite of the high density of ScYLV particles in the cytoplasm. The location of S^– and S⁺ plants close to each other without a discernible pattern of distribution in plots subjected to optimal irrigation and fertilization rule out the possibility that environmental conditions underlay the appearance of symptoms. In plots under severe drought for 3 months, however, all S^– plants become S⁺. Symptom expression did not affect the acid phosphatase activity in the rhizosphere of S⁺ plants.     

Monthly fluctuations during the breeding season of sperm density,volume, motility,and composition of seminal and coelomic fluid in broodfish of Persian sturgeon,Acipenser persicus Borodin, 1897

Obtaining knowledge on the gamete quality of certain species is essential for appropriate management and conservation of wild stocks of the species concerned. In the present study, eighteen breeders (nine males and nine females) of Acipenser Persicus Borodin, 1897 were selected for recording breeding season changes in gamete quality in terms of the parameters: spermatozoa motility indices, sperm volume and density, ionic composition and osmolality of seminal and coelomic fluids. Stripping was performed at the beginning of March and during April and May. Results indicated that sperm volume (ml per male) and density (×10⁹ spz ml^?1) decreased towards the end of the spawning season. There was no significant change in osmolality (mOsmol kg^?1) or in the concentrations (as mm ) of sodium, potassium, magnesium or calcium ions in either the seminal or the coelomic fluids during the breeding season. A significant difference in chloride ion concentration in the coelomic fluid was noted at different times during the spawning season. The percentage of motile spermatozoa at 45 s post‐activation was not significantly different for samples taken at different dates, but the maximum duration of spermatozoa movement, at 15 s post‐activation, was observed in March. This value decreased significantly towards the end of the reproductive season. In conclusion, changes observed in A. persicus sperm parameters during the breeding season suggest an association between such changes and the sperm aging processes.     

CO2‐caused change in plant species composition rivals the shift in vegetation between mid‐grass and tallgrass prairies

Atmospheric CO₂ enrichment usually changes the relative contributions of plant species to biomass production of grasslands, but the types of species favored and mechanisms by which change is mediated differ among ecosystems. We measured changes in the contributions of C₃ perennial forbs and C₄ grasses to aboveground biomass production of tallgrass prairie assemblages grown along a field CO₂ gradient (250–500 μmol mol^?1) in central Texas USA. Vegetation was grown on three soil types and irrigated each season with water equivalent to the growing season mean of precipitation for the area. We predicted that CO₂ enrichment would increase the forb contribution to community production, and favor tall‐grasses over mid‐grasses by increasing soil water content and reducing the frequency with which soil water fell below a limitation threshold. CO₂ enrichment favored forbs over grasses on only one of three soil types, a Mollisol. The grass fraction of production increased dramatically across the CO₂ gradient on all soils. Contribution of the tall‐grass Sorghastrum nutans to production increased at elevated CO₂ on the two most coarse‐textured of the soils studied, a clay Mollisol and sandy Alfisol. The CO₂‐caused increase in Sorghastrum was accompanied by an offsetting decline in production of the mid‐grass Bouteloua curtipendula. Increased CO₂ favored the tall‐grass over mid‐grass by increasing soil water content and apparently intensifying competition for light or other resources (Mollisol) or reducing the frequency with which soil water dipped below threshold levels (Alfisol). An increase in CO₂ of 250 μmol mol^?1 above the pre‐industrial level thus led to a shift in the relative production of established species that is similar in magnitude to differences observed between mid‐grass and tallgrass prairies along a precipitation gradient in the central USA. By reducing water limitation to plants, atmospheric CO₂ enrichment may alter the composition and even structure of grassland vegetation.     

两种鼠蚤在新羽化和吸血后不同时间三种酶的组织化学研究

采用组织化学技术,研究了不等单蚤Monopsyllus anisus (Rothschild)和缓慢细蚤Leptopsylla segnis (Schonherr)新羽化和吸血后24 h、48 h和72 h碱性磷酸酶、酸性磷酸酶和三磷酸腺苷酶的分布和活性。显微摄影及定量图像分析结果显示：新羽化蚤碱性和酸性磷酸酶主要存在于中肠、神经细胞核、精子束头部、精巢附腺、射精管、输卵管和受精囊附腺中;三磷酸腺苷酶各组织中均有分布。吸血消化后,两种蚤中肠3种酶活性均有增强;除碱性磷酸酶在消化72 h 后酶活性有所下降外,其余不同消化时间酶活性增强程度上不存在显著差异。两种蚤卵母细胞发育成熟过程中3种酶活性亦见增强。     

Reproductive guilds (maturation,spawning frequency and fecundity) in the black pomfret,Parastromateus niger (Carangidae) in the Kuwaiti waters of the Arabian Gulf

Oogenesis, oocyte maturation pattern, spawning rhythm, spawning frequency, batch fecundity and oocyte diameter–frequency distribution of the black pomfret, Parastromateus niger (Bloch, 1795) in Kuwaiti waters were investigated from October 2003 to September 2005, using histological and morphological methods. The process of development is divided into four major phases: (i) primary growth phase; (ii) secondary growth phase; (iii) maturation phase; and (iv) spawning phase, followed by the regressed phase. Development of the yolky oocyte is an asynchronous process resulting, by the time of oocyte maturation, in a clear differentiation between a ready batch of oocytes (ready for spawning) and a reserve pool. Consequently, P. niger is capable of spawning multiple times throughout the reproductive season. Spawning frequency estimates, based on final oocyte maturation (FOM) method indicated that the species spawns once every 2.8 days during an 8‐month spawning season lasting from February to September, with a potential annual number of 22.4 spawns. Batch fecundity (BF) (2132–2001 648, mean 406 010 eggs), was significantly positively related to both standard length (SL) (P < 0.05) and ovary‐free body weight (OFBW) (P < 0.05), both parameters being good predictors of BF (r² = 30.8% for SL, from 22 cm onwards, and r² = 29.6% for OFBW, from 129.5 g onwards). No significant differences in monthly BF were found throughout the spawning season. Relative batch fecundity was 336 eggs/g OFBW; thus, estimate for potential annual relative batch fecundity was 7526 eggs g^?1 OFBW. The oocyte diameter–frequency distribution analysis revealed a multimodal distribution (at 100–200, 300–400 and 500–700 μm), confirming the evidence of multiple spawning.