性腺发育不同阶段野生瓯江凤鲚肌肉营养成分的分析与评价

研究旨在针对不同性腺发育阶段(Ⅰ期、Ⅱ期、Ⅲ期、Ⅳ期、Ⅴ期)野生瓯江凤鲚(Coiliamystus)性腺发育情况和肌肉营养成分进行分析与评价。研究表明: 在雌、雄野生瓯江凤鲚性腺从Ⅰ期发育至Ⅴ期的过程中, 成熟系数呈现上升趋势, 凤鲚卵巢的GSI值是精巢GSI值的5倍。在卵巢发育Ⅰ—Ⅴ期, 粗脂肪含量显著下降, 粗蛋白和水分显著上升(P<0.05); 在精巢发育Ⅰ—Ⅴ期, 粗脂肪含量显著上升, 粗蛋白和水分显著下降(P<0.05), 灰分含量先升后降, 说明野生瓯江凤鲚在卵巢发育过程中脂肪为主要供能物质。雌凤鲚肌肉在卵巢发育Ⅰ—Ⅴ期显著上升(P<0.05), 雄凤鲚肌肉氨基酸含量在精巢发育Ⅰ—Ⅴ期显著下降(P<0.05)。但各性腺发育阶段氨基酸组成相对稳定, 雌、雄鱼肌肉总必需氨基酸/总氨基酸分别为(37.88±0.32)%—(41.66±0.44)%和(40.30±0.69)%—(40.94±0.29)%。依据氨基酸评分(AAS)和化学评分(CS)标准, 不同性腺发育阶段的野生瓯江雌、雄凤鲚肌肉中第一限制性氨基酸均为色氨酸, 第二限制性氨基酸均为甲硫氨酸(Met)和胱氨酸(Cys)。在卵巢发育Ⅰ—Ⅴ期, 性腺发育阶段含量最丰富的C16﹕0和C18﹕1呈下降趋势, 可能是作为主要的供能脂肪酸。多不饱和脂肪酸的主要脂肪酸DHA在卵巢发育中呈现先上升后下降趋势。在精巢发育Ⅰ—Ⅴ期, 多不饱和脂肪酸呈现下降趋势。因此, 在繁殖过程中, 野生瓯江凤鲚雌雄鱼的发育特征和肌肉营养组成变化规律及差异与生殖洄游产卵的繁殖习性密切相关。     

光倒刺鲃卵巢发育的初步研究

经激素处理和生态调控的光倒刺鲃,能够完成卵巢发育的全过程。按照形态学特征将卵巢的发育分为6个时期,依组织学特点将卵巢的变化分为5个时期。大多数雌体在3～4龄性腺发育成熟,雌鱼最小成熟为3龄;繁殖期4～9月,为多次产卵类型,繁殖时不能将卵完全产出,完成第一次性成熟并产卵的光倒刺鲃,卵巢逐步由第3期发育至第4期。     

青海湖裸鲤性腺发育的组织学研究

采用石蜡组织切片法,对青海湖裸鲤的性腺发育进行组织学研究,系统地描述了各期卵巢和精巢的形态结构、特征及变化。结果表明:(1)青海湖裸鲤性腺的发育可分为六个时期,卵母细胞从第3时相发育到第4时相基本同步。第Ⅳ期末卵巢,第4时相卵母细胞的卵径大小比较整齐,卵径平均值为2.3mm,第4时相卵母细胞占切面上卵数的85%以上,占切片面积的96%以上,第2、3时相卵母细胞已很少;(2)产后卵巢的组织结构逐步由第Ⅵ期回复到第Ⅱ期,再由第Ⅱ期到第Ⅲ期向第Ⅳ期过渡;(3)性成熟个体有68.12%的雌鱼和83.9%的雄鱼以第Ⅳ期性腺越冬,另有21.01%的雌鱼和10.7%的雄鱼以第Ⅲ期性腺越冬,还有10.87%的雌鱼和5.4%的雄鱼以第Ⅱ期性腺越冬。根据青海湖裸鲤各季节性腺发育情况,作者认为青海湖裸鲤已达到性成熟的个体并不是每年都参与繁殖活动,存在生殖间断性。这反映了青海湖裸鲤的繁殖习性对高原寒冷多变气候的适应性。还对卵黄核和核仁在卵黄形成中的作用以及青海湖裸鲤的产卵类型和生殖间断性进行了讨论。     

南方鲇的繁殖生物学研究：繁殖时间,产卵条件和产卵行为

南方鲇的雌鱼３－４龄性达成熟,雄鱼２—３龄性达成熟;在此期间,同龄组中已成熟个体的体长、体重、肥满度和成熟系数的平均值均大于未成熟个体。繁殖期为３月中旬至５月中旬,其间,嘉陵江平均水温为１５—２４℃。雌鱼的性腺在６－８月由Ⅵ期退化至Ⅱ期,９至１１月经Ⅲ期发育至Ⅵ期,１２月至来年２月以Ⅳ期性腺状况越冬。雌、雄鱼成熟系数周年曲线均为单峰型,该种鱼为单批产卵类型。产卵场为流水卵石浅滩,水深０．４－１．５ｍ,流速每秒０．７ｍ左右。产卵前雌雄亲鱼有相互激烈追逐、咬斗的发情行为．     

大口黑鲈amh基因全长cDNA克隆、表达及多克隆抗体制备

为研究大口黑鲈(Micropterus salmoides)抗缪勒氏管激素(amh)基因的表达及其在性腺发育中的潜在作用,研究利用RACE技术克隆得到了大口黑鲈amh基因,并制备Amh多克隆抗体,通过qRT-PCR、Western Blot分析Amh在大口黑鲈不同组织和不同发育阶段性腺中的表达模式,最后利用HE染色法和免疫组化观察不同发育阶段性腺的形态组织学变化及其与Amh表达的潜在关系。结果显示:大口黑鲈amh基因cDNA序列全长2050 bp,由24 bp5′非编码区、394 bp3′非编码区和1632 bp的开放阅读框组成,共编码543个氨基酸。amh基因mRNA在大口黑鲈11个组织中均有表达,其中雄鱼精巢中表达量最高,肌肉次之,雌鱼卵巢中表达量最高,肌肉次之。amh基因在雌雄鱼不同发育阶段的性腺中表达存在显著差异,精巢中表达量均显著高于卵巢(P<0.05)。同时, Western Blot结果显示Amh蛋白在精巢中表达丰度较高。amh基因在精巢中的表达量呈先上升后下降的趋势,且在孵化后65d鱼精巢中其表达量达到最高(P<0.05),免疫组化结果显示Amh表达于早期精...     

赤眼鳟的繁殖生物学

对赤眼鳟(Squaliobarbus curriculus)的性腺发育、性征、繁殖力、性比、繁殖习性、胚胎发育和胚后发育进行了研究。赤眼鳟初次性成熟年龄为2冬龄。繁殖季节为4月下旬至8月上旬,以5~7月为盛产期。网箱饲养已达性成熟年龄的赤眼鳟性腺成熟系数大于池养及江河野生的。其相对繁殖力为252.8粒/g,高于池养者31.05%,高于野生鱼28.0%。雄鱼体形比雌鱼长,成熟雄鱼在繁殖季节胸鳍具有颗粒状珠星,非繁殖季节珠星消失。雌雄比接近1∶1。赤眼鳟属敞水性产卵类型,为不分批产卵鱼类。广西江河的赤眼鳟天然产卵场有11处,主要分布于郁江,左江和右江各有1处。在水温28~28.5℃的情况下,自受精到孵化所需时间为16 h 10 min,积温453℃.h。鱼苗经17 d发育,鳞片完全,体形、习性与成鱼相似。     

黄尾密鲴性腺发育的研究

生长在湖南地区水库环境的黄尾密鲴,雌、雄鱼性成熟一般为2冬龄。在具备繁殖条件的水库,其黄尾密鲴能自行产卵、受精、繁衍后代,不具备条件的,不能自然产卵,其卵母细胞只能发育到初级卵母细胞阶段(Ⅳ时相),而精细胞则能完成由精原细胞发育到精子的全部发育过程。冬季,达性成熟的黄尾密鲴,雌性卵巢为Ⅲ期,雄性精巢为Ⅳ期。每年的4月份,黄尾密鲴开始达人性腺成熟期,5月中旬至6月初为它的生殖盛期。初级卵母细胞由Ⅲ时相发育到Ⅳ时相基本是同步性的,卵巢成熟系数在一年中只出现1次高峰,经组织学研究与生产实践证明,黄尾密鲴系一年1次产卵类型。     

池养美洲鲥卵巢周年发育和血清激素变化研究

采用形态测量、组织化学以及酶联免疫测定等技术方法,研究了淡水池养美洲鲥卵巢发育和促性腺激素(LH和FSH)和性类固醇激素(T和E2)的年周期变化规律。结果表明,美洲鲥产卵类型属非同步分批产卵,按组织学特点其卵母细胞发育可分为5个时相,依据形态特征其卵巢发育可划分为相应的5个时期。在性腺发育过程中,雌鱼性腺指数(GSI)和肝脏指数(HSI)没有出现明显的变化(p0.05);LH在第Ⅴ时期的表达量显著性低于第Ⅱ、Ⅲ和Ⅳ时期的表达量(p0.05),其他各激素在不同时期均没有明显差异(p0.05)。不同激素间的两两相关性表明血清中LH水平和FSH水平存在显著正相关关系(r=0.613,p0.05),FSH水平和T水平显著负相关(r=-0.605,p0.05),LH水平和T水平显著负相关(r=-0.699,p0.05)。Person相关系数表明卵巢性腺指数和肝脏指数同几种激素之间没有显著的相关性(p0.05)。通过与野生美洲鲥以及其他一些硬骨鱼类卵巢发育规律比较表明:在目前养殖条件下美洲鲥雌鱼营养积累不足,其性腺发育调控受到影响。     

长吻鮠的卵巢发育和周年变化及繁殖习性研究

本文报道了长吻卵巢发育与周年变化特点和繁殖习性。依外形和组织学特征,性成熟前至性成熟鱼卵巢的发育可分为卵原细胞期、单层滤泡期、卵黄泡期,卵黄充满期、成熟期和退化期６个时期,Ⅰ期性腺只存在于１龄内个体,Ⅱ期性腺持续２年左右,性成熟前发育至Ⅲ期,经Ⅳ、Ⅴ期发育至成熟并首次参与繁殖,最小性成熟个体３龄。性成熟后个体卵巢的发育只有后５个时期。繁殖期４—６月,产后卵巢很快退回到Ⅱ期,Ⅲ期卵巢越冬。一次产卵类型。卵壳膜源于滤泡细胞。卵粒的退化吸收分５步完成。     

两种方式对红螯螯虾同步产卵影响的研究

对红螯螯虾采取雌雄分养和烫伤雌虾单侧眼柄两种方法进行处理,研究其对红螯螯虾卵巢发育和产卵率的影响,进而判断对同步产卵的影响。结果显示,雌雄分开放养对雌虾性腺发育有促进作用,其性腺指数显著高于幼虾刚离体的雌虾和试验结束时未产卵的雌虾,而肝胰腺指数低于试验初的雌虾,但14 d的单放时间偏长;单养14 d组和烫伤单侧眼柄组的产卵率分别为35%和50%,小于对照组(75%);雌虾交配前不需要蜕壳;幼虾离体的雌虾不再次交配也可以产卵,但不一定能孵出苗。研究表明,雌雄分养和烫伤雌虾单侧眼柄对于刚理出的产空雌虾的同步产卵促进作用不明显,生产上对于幼虾离体后的雌虾应立即雌雄配组。     

Annual reproductive cycle of a bitterling, Tanakia tanago, reared in an outdoor tank

From June 2000 to September 2001, we investigated the presence of eggs spawned in Margaritifera laevis and the seasonal changes in the gonads of Tanakia tanago. Eggs were observed from mid-March to mid-September. In females with a shrunken ovipositor, as the GSI gradually increased, most ovaries were in the prespawning phase (Oct-Mar). As the GSI increased further, most ovaries were in the early spawning phase (Mar-Jun). As the GSI gradually deceased, ovaries in the late spawning phase appeared (Jun-Sep). When the GSI was very low, most ovaries were in the postspawning phase (Sep-Oct). In males, when the GSI was low, most testes were in the early prespawning phase from Oct-Dec. As the GSI gradually increased, most testes were in the late prespawning phase (Dec-Jan). As the GSI increased further, testes were in the early spawning phase (Jan-Jun). As the GSI gradually decreased, amost testes were in the late spawning phase (Jun-Sep). When the GSI was very low, most testes were in the postspawning phase (Sep-Oct). These results indicate that T. tanago has a distinct annual reproductive cycle and is a spring-autumn spawner. Based on the relationship between reproductive activity and environmental factors, the spawning season of T. tanago appears to be initiated by increasing temperature and / or longer days in spring and to be terminated by shorter days in autumn.     

Reproductive biology of the East African lungfish (Protopterus aethiopicus) in Mwanza Gulf, Lake Victoria

The population of lungfish (Protopterus aethiopicus (Heckel)) was sampled by bottom trawling for 12 months from June 1984 to May 1985. A total of 576 lungfish were investigated. The sex ratio in the population was 1 male : 1 -98 females. Average size at maturity was found to be 960cm total length (TL) but the smallest matured male measured 72-9 cm and the female 840 cm TL. It was not possible to establish spawning times and reproductive activity trends either from the variation in relative condition factor ‘kn’ or from gonadosomatic index (GSI). Whereas male lungfish seem to show no further increase in GSI after gonadal maturity stage III, females exhibit GSI values that vary directly with oocyte maturation and ovarian weights. Fat deposited along the gonads was found to be a better basis for assessing maturation and spawning in lungfish. The potential spawning period was found to be from September to May. Absence of lungfish with spent gonads over the 12-month period and the presence of different egg sizes in ovaries, suggest that the fish is a multiple spawner. Fecundity was found to range from 705 eggs (in 84-cm TL fish) to 14,922 eggs (in 130-cm TL Fish).     

Annual cycles of growth and reproduction in hatchery–reared Florida largemouth bass, Micropterus salmoides floridanus, raised on forage or pelleted diets

Annual cycles of growth and reproduction of hatchery–reared Florida largemouth bass, Micropterus salmoides floridanus , were investigated. Animals were raised on either forage (goldfish, Carassius auratus ) or a pelleted salmon feed. Male and female year–class 1 largemouth bass were sampled throughout one complete yearly cycle (January–December). A biphasic growth cycle was observed in both forage–fed and pellet–fed fish. No increase in body length or weight was observed until approximately midway through the spawning period (May), after which fish grew at a consistent rate for the remainder of the study. The reproductive cycle of forage–fed fish was characterized by a rapid increase in gonadosomatic index (GS1) between January and April, followed by a prolonged spawning period (April–July) during which GSI progressively declined. Fully regressed gonads were observed in September and October, and a resumption of gonadal recrudescence was observed between October and December. Visceral adipose deposits (expressed as mesenteric fat index; MFI) were resorbed during gonadal growth and the initial stages of the spawning period, and restored during the post–spawning phase. Fish raised on pelleted feed had growth and reproductive cycles that parallelled those of forage–fed fish, but several significant differences were observed between the two diet groups. During the growth phase of the cycle, pellet–fed largemouth bass grew significantly faster than forage–fed largemouth bass, and had significantly larger MFIs than forage–fed largemouth bass at all times of the year. Pellet–fed fish also had significantly larger GSIs than forage–fed fish. These data indicate that diet composition may be an important determinant of growth and reproductive function in this species.     

雅鲁藏布江黑斑原鮡繁殖生物学研究

对2004-2006年采集于雅鲁藏布江拉萨河的190尾黑斑原鮡进行了繁殖生物学研究。雄性最小性成熟(精巢Ⅳ期)个体体长141.7mm,体重45.2g,性体指数1.09%,雌性最小性成熟(卵巢Ⅳ期)个体体长146.8mm,体重66.7g性体指数11.52%,相应年龄均为5龄。初次性成熟年龄(L50):♂,170.1mm相应年龄为7龄;♀,150.2mm,相应年龄5龄。通过组织切片法和GSI的周年变化分析,繁殖时间集中在5-6月,每年繁殖一次,繁殖之后的6-8月卵巢从Ⅵ期回复到Ⅲ期,9月卵巢发育到Ⅳ期越冬。卵径频率分布显示,卵巢发育类型为分批同步型,卵巢中至少存在2批卵径,每年成熟一批卵并同时产出,产卵类型为完全同步产卵。卵黏性,成熟卵卵径在2.04-3.37mm之间,平均(2.83±0.16)mm。对19尾产卵前夕(体长为151.0-210.0mm)的标本进行统计,其绝对繁殖力范围在525-2058粒之间,平均为(1244±346)粒,相对繁殖力为(14.7±5.8)粒/g。绝对繁殖力与体长呈直线正相关,表达式为F=13.624L-1187。       

Role of the tidal cycle in the gonadal development and spawning of the tropical wrasse Halichoeres trimaculatus

Many wrasses on coral reefs exhibit daily spawning that peaks around daytime high tides. In this study, we examined tidal-related ovarian development in the threespot wrasse, Halichoeres trimaculatus, a species common on coral reefs in the Indo-Pacific Ocean. When the fish were collected in the morning at different tidal phases, the gonadosomatic index (GSI) and ovarian histology changed; concomitant with increases in GSI towards high tide, a clutch of the most advanced oocytes developed from vitellogenic to maturation stages. Ovulated eggs and post-ovulated follicles (POF) existed in most ovaries around high tide, but only POF remained around ebb tide, suggesting that spawning occurred during or after high tide. We noticed that tidal-related spawning was considerable in the morning and that most ovaries collected on the afternoon high tide exhibited post-spawning features. This suggests that certain labrid species possess plasticity with regard to their spawning time and utilize potent environmental cues to ensure their reproductive success. When pieces of ovary were incubated with precursor steroids, high conversion of testosterone to 17beta-estradiol occurred during high and ebb tides, while that of 17alpha-hydroxyprogesterone to 17alpha,20beta-dihydroxy-4-pregnen-3-one and 17alpha,20beta,21-trihydroxy-4-pregnen-3-one was observed during low and flood tides. Incubation of pieces of ovary with human chorionic gonadotropin resulted in similar fluctuations in the steroid hormones with tidal phase. Production of these steroid hormones correlated with oocyte development in the ovaries and was probably regulated by gonadotropin. These results demonstrate that the daily cycle is fundamental for oocyte development, and that the tidal cycle is superimposed on this process.     

Sexual maturation in female triploid plaice, Pleuronectes platessa, and plaice × flounder, Platichthys flesus, hybrids

The gonads of immature female triploid plaice ( Pleuronectes plafcssa ) and plaice × flounder ( Platichthys flesus ) hybrids produced from cold-shocked eggs were examined. They were less than half the size of those in diploids of similar age and contained only small numbers of developing oocytes. These were similar both in size and cytological appearance to those in the controls. The bulk of the ovarian tissue was composed of nests of small undifferentiated cells resembling the oogonia in the ovaries of diploid fish. Mature, 5 year old triploid hybrids which had not produced eggs over the previous two spawning seasons were killed in January when the diploid controls were nearing spawning condition. The ovaries of these fish were smaller than those of the controls and contained degenerating oocytes at maturation stage V. The advantage of sterile female triploid fish for fish-cultivation is discussed.     

Estimating the spawning activity of fish species using nuclear and mitochondrial environmental DNA concentrations and their ratios

1. During spawning activity, fish release large amounts of sperm and eggs into the water, which has been assumed to cause an increase in environmental DNA (eDNA) levels and nuclear DNA/mitochondrial DNA ratios. To test whether these assumptions are valid and whether nuclear and mitochondrial eDNA analysis can be used to monitor the spawning activity of freshwater fish, we conducted field eDNA surveys and traditional surveys using common carp (Cyprinus carpio), largemouth bass (Micropterus salmoides) and bluegill sunfish (Lepomis macrochirus) as model species.
2. Fish spawning periods were estimated based on age, as estimated using the body lengths of juveniles collected in the Miharu reservoir in Fukushima, Japan. The results showed that the main spawning periods of largemouth bass and bluegill sunfish were from April to July and from July to August, respectively.
3. Field eDNA surveys were conducted in the Hebisawagawa front reservoir, which is connected to the Miharu reservoir. From March to August 2019 and 2020, weekly eDNA sampling was conducted at three sites, and daily sampling was conducted at six sites from 23 June to 3 July 2020. The eDNA concentrations of the nuclear internal transcribed spacer 1 (ITS1) and mitochondrial cytochrome B (CytB), as well as the ITS1/CytB ratio, were measured for each of the three fish in each water sample. Water temperature had a statistically significant effect on eDNA concentration, probably reflecting the relationship between water temperature and spawning.
4. We created generalised additive mixed models to estimate spawning activity periods based on weekly eDNA data. The estimated periods of spawning activity for common carp, largemouth bass and bluegill sunfish were March to May, May to July, and May to August, respectively. The estimated spawning periods coincided with known fish ecology or the results of traditional methods. This method also has been applied to daily eDNA samples, showing the feasibility of high-resolution estimation of spawning activity.
5. For common carp and bluegill sunfish, we were able to estimate the spawning period using this method. Although the method is affected by biomass and the diffusion and degradation of eDNA, it has the potential to accurately estimating spawning activities. These then can be estimated without conducting laborious traditional surveys, facilitating the monitoring of reproduction by rare, invasive or important fishery species. Further research on the diffusion distance and degradation time of the eDNA concentration peak caused by fish spawning activity may improve the accuracy of monitoring.

     

Plasma sex steroids and gonadosomatic index variations during ovarian development of female wild yellowfin seabream (Acanthopagrus latus)

The yellowfin seabream (Acanthopagrus latus) has a good potential for mariculture in Iran, so studies on various aspects of its culture has been carried out. In this study, the characteristic of reproduction in this species was investigated. The involvement of steroid hormones in the development of gonads was assayed by sampling blood and gonad tissue in 85 females of A. latus in fish caught from the Musa Creek, north-west Persian Gulf from October 2010 to May 2011. Gonadosomatic index (GSI) and hepatosomatic index (HSI) were recorded. Developmental stages were associated with changes in sex steroids, GSI and HSI. GSI and HSI began to increase in November (immature stage) and reached the highest value in March (ripe stage), decreasing sharply in April (spent stage). Plasma values of 17β-estradiol (E2) levels reached their highest levels in spawning females in March and then decreased in April. Progesterone levels were low through ovarian development, reached a maximum simultaneously with testosterone in February (mature stage) and then decreased in March (ripe stage). The seasonal changes observed in plasma steroid levels and GSI reflected the pattern of oocyte development and the spawning behavior of yellowfin seabream and were typical of the patterns described in most multiple spawners studied to date.     

饲料中添加牛乳铁蛋白肽对大口黑鲈幼鱼生长性能、消化酶活力、肠道组织结构及抗病力的影响

为探讨饲料中添加不同水平牛乳铁蛋白肽(Bovine lactoferricin, LfcinB)对大口黑鲈(Micropterus salmoides)幼鱼生长性能、消化酶活力、肠道组织结构及抗病力的影响,试验选取了初均重为(19.88±0.03) g的450尾大口黑鲈,随机分成5组,每组3个重复,分别在基础饲料中添加0(阴性对照组)、1000、1500和2000 mg/kg牛乳铁蛋白肽,并设置基础饲料+30 mg/kg氟苯尼考为阳性对照组,共5种试验饲料。试验周期为8周。结果表明:(1)随着牛乳铁蛋白肽添加量的增加,大口黑鲈的末均重(FBW)、增重率(WGR)和特定生长率(SGR)均呈现先升高后降低的趋势, 1000 mg/kg牛乳铁蛋白肽组大口黑鲈的生长性能最好且与对照组(阴性对照、阳性对照)对应数据差异显著(P<0.05)。(2)与两个对照组比, 1000 mg/kg牛乳铁蛋白肽组大口黑鲈的肠道胰蛋白酶、α-淀粉酶和脂肪酶活力均显著上升(P<0.05)。(3)与两个对照组比,饲料中添加1000 mg/kg牛乳铁蛋白肽可显著提高大口黑鲈前肠、中肠及后肠的绒毛高度和宽度(...     

Gonadal development and non-functional protogyny in a coral-reef damselfish, Dascyllus albisella Gill

All gonads of the Hawaiian dascyllus Dascyllus albisella , irrespective of the final sex of individuals, developed an ovarian lumen and primary-growth-stage oocytes after an initially undifferentiated state. From this ovarian state or from more differentiated ovaries, some gonads redifferentiated into testes. None of 117 individuals examined had a gonad containing degenerating vitellogenic oocytes and proliferating spermatogenic tissue. Eleven individuals had gonads containing degenerating cortical-alveolus-stage oocytes and developing spermatogenic tissue. The size of these individuals overlapped with the female size range in which the majority of the females were still in the middle of the maturation process. They were absent from the larger size range where the majority of females had vitellogenic oocytes. This indicated that the transition toward maleness is likely to have occurred after the onset of cortical-alveolus stage, but before final oocyte maturation and spawning as females. Therefore the protogynous pattern of gonadal development was non-functional. There was no dimorphism in the sperm duct configuration, and all the testes were secondary testes reported for diandric, protogynous species with undelimited gonads. Very early development of an ovarian lumen appeared to have resulted in a secondary-male configuration in all testes, although redifferentiation into males appeared to have occurred before sexual maturity and spawning as females.