Growth and reproduction of the Japanese mantis shrimp, <Emphasis Type="Italic">Oratosquilla oratoria</Emphasis> (De Haan 1844) in the coastal area of Tongyeong,Korea

Growth and reproduction of the Japanese mantis shrimp, Oratosquilla oratoria, were investigated in the Tongyeong, Korea from July 2014 to August 2015. A total of 2,621 samples (1,380 females and 1,241 males) were collected during the study period. Females were observed more frequently than males. The mean body length (BL) was 128.5 ± 0.38 mm in females and 126.9 ± 0.42 mm in males. The mean body weight (BW) was 31.2 ± 0.28 g in females and 31.1 ± 0.32 g in males. There was a significant difference in the length-frequency distribution between females and males. The relationship between BL and BW was lnBW = 2.85 × lnBL - 10.43 for females and lnBW = 2.87 × lnBL -10.52 for males. The gonadosomatic index (GSI) varied on a monthly basis. The GSI reached a maximum in May and a minimum in November. The highest values of the GSI coincided with the spawning period of O. oratoria. Larger individuals of O. oratoria have their spawning season earlier than smaller ones. The size at sexual maturity of females was estimated as 96.5 mm. The Von Bertalanffy growth function parameters were BL_∞ = 184.5 mm, K = 0.72 year^?1, C = 0.36 and WP = 0.45 for females and BL_∞ = 183.75 mm, K = 0.82 year^?1, C = 0.38 and WP = 0.22 for males. The growth of males was slightly faster than females. The present study will help with the fisheries management of O. oratoria based on ecological parameters.     

Gonadal development in Japanese medaka (Oryzias latipes) exposed to 17 β-estradiol

The purpose of this research was to evaluate the Japanese medaka (Oryzias latipes) as an ecotoxicological model for the rapid evaluation of environmental estrogens. A novel short-term (48-h) exposure to 17 β-estradiol is proposed in development of a positive control for disruption of gonadal development. Recently hatched medaka fry (30 fry per dose group) with undifferentiated gonads were exposed to 4.0, 29.4, and 115.6 μg/liter of 17 β-estradiol (acetone carrier) for 48 h in a water bath at 25 °C. The fry were then grown-out in spring water for 2 weeks, killed, and processed for histological evaluation. High lethality was encountered during the grow-out period in the 115.6 μg/liter dose group. Fry in the spring water and acetone (carrier) control groups developed into females or males. Fry exposed to 17 β-estradiol developed primarily into females or had testis-ova.     

Reproductive cycle,size at first maturation and fecundity in the golden ling,Genypterus blacodes,in Chile

Reproductive aspects of the golden ling, Genypterus blacodes, from the southern Chilean fishery are presented. Gonad samples were obtained from commercially caught fish between September 1999 and October 2000. Stage of sexual maturity, gonadosomatic index (GSI), and size of females at 50% maturation determined by logistic methods and by plotting the mean GSI by total length, are presented for both years. Batch fecundity was calculated from subsampling mature ovaries containing oocytes in advanced stages of maturity. Changes in GSI and monthly diameters of oocytes showed that ling had a spawning season which extended between August and November. The sizes at first maturity in 1999 and 2000 were 86.6 and 84.8 cm total length (TL), respectively, using the logistic model, and 84 cm for both years using the mean GSI‐size plotting method. The mean batch fecundity was 333 330 oocytes per individual for fish sized 85–120 cm TL. Fecundity increased with increasing weight and total length.     

Reproductive traits of sandbird octopus, <Emphasis Type="Italic">Amphioctopus aegina</Emphasis> (Gray, 1849) from Mandapam coastal waters (Palk Bay), Southeast Coast of India

The sandbird octopus Amphioctopus aegina (Gray, 1849) is one of the important octopod species in trawl catches in Mandapam waters (Palk Bay). The reproductive biology of this species from these waters was studied from October 2001 to September 2002. In the majority of months(Jan–June), the sex ratio was biased towards males. The ratios of males to females increased consistently with respect to weight Total weight at first maturity were 78.78g for females and 40.8 g for males. Four maturity stags were recognized for females and two for males. Maturation and spawning occur all year round, with a peak during October and another peak during January–February. In males, no definite seasonal changes were observed in gonadosomatic index (GSI) values. In females there were two peaks in GSI values during October and January–February. For individuals of a DML range of 67–85 mm fecundity varied between 2,962–8,820 oocytes. The average relative fecundity was estimated at 68 to 83 and the average number oocytes per gram of ovary were 488 to 539.     

菲、芘、苯并(a)芘单一暴露及分别与α-萘黄酮(ANF)联合暴露对海水青鳉(Oryzias melastigma)胚胎发育毒性效应的比较研究

为了解和探讨3~5环PAHs对海水鱼类胚胎发育的毒性及作用方式,比较研究了菲(phenanthrene,Phe)、芘(pyrene,Py)、苯并(a)芘(benzo(a)pyrene,BaP)单一暴露和三者各自与α-萘黄酮(α-naphthoflavone,ANF)联合暴露对海水青鳉(marine medaka, Oryzias melastigma)胚胎发育的毒性效应。胚胎体内EROD活性、发育畸形、孵化率和心律等毒性指标被测定,结果显示:Phe,Py和BaP对海水青鳉胚胎体内EROD活性的诱导能力大小为BaP>Py>Phe,各化合物对EROD诱导与发育畸形之间的关系较为复杂,除Phe所引起的EROD诱导与畸形指数之间呈显著相关(r=0.95,p=0.015)外,Py和BaP均无相关性;在100 μg/dm3 ANF影响下,CYP1A活性诱导被抑制,但胚胎发育的畸形指数被显著提高,ANF分别与Phe,Py和BaP的联合暴露对胚胎发育呈潜在的协同作用。本文研究初步表明,3~5环PAHs化合物对海水青鳉胚胎发育的毒性作用方式可能不同;CYP1A活性抑制在PAHs混合物对海水青鳉胚胎发育的毒性作用过程中未起到缓解毒性的作用,CYP1A抑制剂与PAH型CYP1A诱导剂的混合物对鱼类胚胎发育具有潜在的协同毒性作用,现有的PAHs混合物毒性风险评价方法可能低估了实际环境中PAHs的风险;海水青鳉早期生活阶段的心脏发育对PAHs混合物暴露较为敏感,可推荐其作为生物标志物指示PAHs或溢油污染。     

Effect of parental exposure to trenbolone and the brominated flame retardant BDE-47 on fertility in rainbow trout (Oncorhynchus mykiss)

We exposed sexually maturing male rainbow trout (Oncorhynchus mykiss) to BDE-47 (a polybrominated diphenyl ether) and female rainbow trout to trenbolone (an anabolic steroid). Male trout were orally exposed for 17 days to 55 microg/kg/day BDE-47 and female trout continuously exposed for 60-77 days to a measured trenbolone water concentration of 35 ng/L. After the exposure, eggs and semen were collected and in vitro fertilization trials performed using a sperm:egg ratio of 300,000:1. In the BDE-47 study, eggs from control females were fertilized with semen from exposed males, while in the trenbolone study, eggs from exposed females were fertilized with semen from control males. All treatments were evaluated at two-three early developmental time-points representing first cleavage (0.5 day), embryonic keel (9 days), and eyed stages (19 days), respectively. The results indicated that BDE-47 exposure did not alter fertility as embryonic survival was similar between control and exposed groups. Trenbolone exposure also did not alter embryo survival. However, in the embryos fertilized with eggs from trenbolone exposed females, a noticeable delay in developmental progress was observed. On day 19 when eye development is normally complete, the majority of the embryos either lacked eyes or displayed under-developed eyes, in contrast to control embryos. This finding suggests steroidal androgen exposure in sexually maturing female rainbow trout can impact developmental timing of F1 offspring.     

Exposure to environmentally relevant concentrations of different nonylphenol formulations in Japanese medaka.

The time course of exposure to p-nonylphenol (NP) from two different sources was compared to equivalent exposures of 17-beta-estradiol (E2) and a solvent control (ethanol; EtOH). Japanese medaka were exposed for 4 days to a nominal concentration of 20 micrograms/l of either NP-I (Schenectady International, Inc.), NP-II (Aldrich), or E2, and were then placed in untreated water for 5 days. Tissue samples were taken at two time points during the 4-day exposure and two time points during the 5 days following exposure. Liver homogenates were analyzed using a western blot to detect vitellogenin (VTG) and quantified by measuring the optical density for each lane. Preliminary results indicate that E2 significantly increased VTG staining above the level observed in EtOH-treated controls for both males and females. A two-way analysis of variance (ANOVA) indicates that NP from both sources, as well as E2, significantly increased VTG staining in males (ANOVA, n = 48, P < 0.001; Tukey pairwise tests, all P < 0.008). A significant increase in VTG was observed in E2-treated males and females the first day following transfer into toxicant-free water (two-way ANOVAs, both n = 48, P < 0.003; Tukey pairwise tests, all P < 0.019). If confirmed, this extended response observed for low-level exposures may represent a significant factor for sampling scenarios following pulsitile exposure.     

Developmental evaluation of a potential non-steroidal estrogen: triclosan.

Triclosan is an antibacterial agent commonly used in industry and often detected in waste-water effluent. The potential of triclosan to act as an endocrine disruptor was examined because its chemical structure closely resembles known non-steroidal estrogens (e.g. DES, bisphenol A). Japanese medaka fry (Oryzias latipes) were exposed for 14 days beginning 2 days post-hatch to triclosan (100, 10, 1 micrograms/l), 17-beta estradiol (E2; 1 microgram/l), or a solvent control (ethanol). Two months post-exposure, the phenotypic sex of each adult was assessed visually using sexually dimorphic fin shape and size. The proportion of females in each group was similar for triclosan-exposed animals and solvent-treated controls (ethanol 53%, 1 ppb 58%, 10 ppb 45%, 100 ppb 36%) although E2 treatment did produce 92% female adults. Sexually dimorphic fin traits were quantified to look for potential effects of triclosan and E2 on the development of secondary sexual characters. These results do not support the hypothesis that triclosan is potently estrogenic. However, changes in fin length and non-significant trends in sex ratio suggest triclosan is potentially weakly androgenic.     

Influence of exposure history on vitellogenin induction in medaka: a case of ‘estrogen memory’?

In this study, we exposed female medaka to aqueous solutions of o,p′-DDT until tissues residues reached approximately 100 g/g. Male offspring of exposed females and unexposed males that were raised to 6 months of age differed from untreated controls in their response to subsequent 4-day static exposure to 17-β-estradiol (E2; 12 μg/l as determined by steroid EIA). Homogenized liver samples from males from DDT treatments, females from DDT treatments, control males and control females produced two vitellogenin bands on a Western blot with no qualitative differences in band size and location between the groups. To quantify the relative level of vitellogenin, 5 μg of protein from each sample was analyzed by slot blot. A statistical comparison revealed a significant difference in optical density of the slots between control males and DDT exposed males (analysis of variance, n=18, P=0.021; Tukey test, n=9, P=0.047) indicating that previous transgenerational exposure to DDT enhanced the vitellogenic response to E2 exposure in males. These results indicate that exposure to estrogenic chemicals may produce an ‘estrogen memory’ which potentiates the response to a subsequent exposure event.     

Phenotypic and genetic differentiation in young-of-the-year common sole (Solea solea) at differentially contaminated nursery grounds

Growth-related characters, condition factor, and genetic differentiation were investigated for a single cohort of young-of-the-year (YOY) sole within and among nurseries with differing levels of heavy metals (Cd, Cu and Zn) contamination in the two Charentais Straits, Bay of Biscay, France. Analyses were performed when individuals recruited (May), then after a full summer spent in each nursery (October). Levels of phenotypic and genetic diversity were compared, together with genetic differentiation at a candidate metallothionein (MT) locus and three putatively neutral microsatellite loci. No phenotypic or genetic differentiation was detected among nurseries in May, but significant variation at each phenotypic trait and at the multilocus level in October (P < 0.001). Single locus analysis demonstrated that only the MT locus was significantly differentiated among nurseries, whether corrected for null alleles or not (θ=0.0401 and θ(corr.FreeNA)=0.0326, respectively; P < 0.001). Results indicate that phenotypic differences among YOY sole nurseries present a molecular correlate acting at identical spatio-temporal scales among nurseries, potentially reflecting differential selective pressure among nurseries in response to contamination.     

Determining the sensitive developmental stages of intersex induction in medaka (Oryzias latipes) exposed to 17 beta-estradiol or testosterone.

Certain environmentally persistent compounds can adversely affect reproduction by acting as steroid hormone agonists or antagonists. The goal of the present study was to determine the developmental stage most susceptible to exogenous hormone (estradiol and testosterone) exposure using a small teleost model. In the first (pilot study) of two experiments, medaka (Oryzias latipes), at varying developmental stages, were bath-exposed to 5 micrograms/l 17 beta-estradiol for 24 h. At 5 months of age, fecundity, fertility and embryo and larval viability (reproductive success) were investigated in control and exposed groups. Fish at 1, 1.5, 2 and 5.5 months of age were also sampled, processed and examined histologically for gonadal alteration. No significant differences in mortality, gonadal morphology, body weight, sex-ratio or time to maturity were seen between control and exposed fish. At 5 months, however, when exposure groups were compared to controls, significant differences were seen in reproductive success and viability of offspring. A second experiment exposed embryo stage 10, and 1-, 7- and 21-day-old larvae for 6 days to 15 micrograms/l 17 beta-estradiol or 100 micrograms/l testosterone. No significant differences were seen at 5 months in mortality, body weight, or time to sexual maturity. However, sex-ratios were significantly biased toward female in the stage 10, 1- and 7-day post-hatch estradiol exposure groups. No significant changes in sex-ratio were associated with testosterone exposure at any developmental stage. Further, intersex gonads were observed in fish from all groups exposed to 15 micrograms/l estradiol. Only those fish exposed as newly hatched fry or at 1 week post-hatch displayed intersex gonads following 100 micrograms/l testosterone exposure. Data from these experiments show that newly hatched fry are that life stage most sensitive to hormone exposure and the most appropriate to use in determining effects of known endocrine-disrupting compounds.     

Gonadal cycle of the dwarf oyster Ostreola stentina from the south of the Gulf of Hammamet on the eastern coast of Tunisia

The reproductive cycle of the dwarf oyster Ostreola stentina (Payraudeau 1826) at Monastir in the south of the Gulf of Hammamet on the east coast of Tunisia was studied using condition indices and histological techniques. The species was shown to be a protandric hermaphrodite. Whereas males dominated the small shell length categories (12.45–34.99 mm), females (21.00–47.16 mm) and hermaphroditic individuals (30.00–57.95 mm) become more predominant with increasing size, suggesting that individuals may undergo protandric development. The overall male to female ratio was 1.0:1.7. Gametogenic development begins in February–March for females and in September for males, and spawning occurs during April and July for females and December and July for males. There was a clear relationship between the gonadal condition index and gametogenic stage for both sexes. Values increased from zero in their early stage of development and peaked during ripe and spawning stages, before decreasing in the spent stage. However, the meat condition index did not show any statistical differences between gametogenic stages.     

Suppressive effects of benzo[a]pyrene upon fish immune function: evolutionarily conserved cellular mechanisms of immunotoxicity

Knowledge gained through the use of alternative animal models has significantly enhanced our understanding of life at all levels of biological organization. The discipline of toxicology is under considerable pressure to develop such models due to increasing public concern regarding the experimental use of mammals. Studies in this laboratory have focused on the utility of a small laboratory fish model, the Japanese medaka (Oryzias latipes), to investigate immunotoxicological effects of benzo[a]pyrene (BaP). BaP is a ubiquitous environmental contaminant and known mammalian immunotoxicant. This laboratory has demonstrated that in vivo exposure of medaka to BaP (2-200 microg/g BW) significantly depresses both innate and humoral immunity. Further studies have indicated that BaP activates its own biotransformation pathway within medaka immune cells following both in vivo and in vitro exposure. In addition, reduction of BaP metabolism with alpha-naphthoflavone results in the reversal of BaP-induced suppression of antibody production in vitro. Inhibition of CYPlA-mediated metabolism within medaka immune cells also alleviates the immunotoxicity induced by benzo[a]pyrene-7,8-dihydrodiol, but not benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE). This suggests that BPDE may be an ultimate immunotoxicant. Results from this study in medaka are in agreement with previously conducted rodent studies that indicated a role for immunotoxic BaP metabolites in BaP-induced suppression of humoral immunity.     

疣荔枝螺(Thais clavigera)性畸变现象的解剖学和组织学研究

以解剖学和组织学的方法研究了有机锡污染生物指示种——疣荔枝螺的正常雄、雌和性畸变个体的生殖系统。结果表明，性畸变个体除具有正常的雌性器官外，还有输精管或阴茎。当性畸变程度严重时，性畸变不育个体的卵囊腺组织出现破损，并具有不规则块状的不育卵囊和未排出的正常卵囊。虽然性畸变个体的雄性器官具有表现出雄性功能的潜能，但由于缺少精巢或完整的前列腺，不可能发展成为具有雄性功能的个体，这与性逆转有着本质上的区别。有机锡污染引起的性畸变已对疣荔枝螺种群生存构成了潜在的威胁，深化此类负面生态效应的研究是人类面临的重要课题。     

Induction of vitellogenin in vivo and in vitro in the model teleost medaka (Oryzias latipes): comparison of gene expression and protein levels

Quantification of the egg yolk precursor vitellogenin (VTG) in fish has become a standard technique to detect estrogenic effects of known chemicals and environmental samples. In the present study, we have analysed VTG induction by estradiol, ethynylestradiol and genistein exposure in the model teleost medaka (Oryzias latipes) and demonstrate that the medaka is a suitable model system to analyse estrogenic effects. By comparing VTG gene expression and protein levels we show that in principal both techniques can be used to study VTG induction in vivo (juvenile and adult males) and in vitro (primary cultures of male liver cells). If a short term in vivo or in vitro exposure is performed, detection of mRNA might be sufficient. For long term studies with the need to detect weak estrogenic chemicals and a precise quantification, immuno-chemical detection may be favoured.     

Expression profiles of sex-related genes in gonads of genetic male Takifugu rubripes after 17β-estradiol immersion

Estradiol treatment during early life stages of tiger puffer Takifugu rubripes induces feminization in genetic males.However,the ovaries in genetic males may revert to testes once estradiol treatment is halted.Therefore studies should investigate molecular mechanisms underlying ovary-to-testis recovery in genetic males after treatment.In the present study,tiger puffer were exposed to 10,and 100μg/L 17β-estradiol(E 2)from 15 to 100 days post-hatching(dph),then gonad phenotypes and expression profi les of six sex-related genes(cyp19a,foxl2,dmrt1,amh,sox9a,and sox9b)were characterized after the exposure.Results showed that both 10 and 100μg/L E2 induced ovarian development in genetic males at 100 dph.However,all ovaries induced by 10μg/L E2 first developed into intersexual gonads and subsequently reverted to testes after the exposure.As for treatment of 100μg/L E2,while the rest of the ovaries maintained morphological stability,percentages of intersexual gonads reached 38%-57%,and none were reverted to testes.Increased mRNA levels of cyp19a,foxl2 and sox9b and decreased mRNA levels of dmrt1,amh,and sox9a were observed during the ovarian development in genetic males.While contrary gene expression profiles were detected during ovary-to-testis transformation.The mRNA levels of all the six genes were increased during the development of intersexual gonads.These results indicated that up-regulation of dmrt1,amh and sox9a is associated with initial ovary-to-intersexual transformation,and suppression of foxl2,cyp19a and sox9b is essential for complete ovary-to-testis recovery in genetic males.This research will help to trace the molecular processes underlying gonadal transformation in teleosts.     

Reproductive biology of the leatherjacket,Meuschenia scaber (Monacanthidae) (Forster 1801) in the Hauraki Gulf,New Zealand

The leatherjacket Meuschenia scaber is widely distributed in Australasian waters, and is a valued bycatch of inshore bottom trawl fisheries although little is known of its life history. Here, we describe the reproductive biology of the species based on 651 leatherjackets sampled in the Hauraki Gulf, New Zealand, between July 2014 and March 2016. The maximum total length (L_T) recorded for females and males were 320 and 315?mm, respectively, with both sexes present in all size classes. Monthly analysis of gonad condition revealed a clear spawning season from late austral winter to early summer (August–December), and histological analysis of the ovaries revealed that M. scaber is an indeterminate serial spawning gonochorist. The estimated sizes at sexual maturity (L₅₀) for females (189.9?mm L_T) and males (188.4?mm L_T) did not differ significantly. Relatively small testes, sexual dimorphism and underwater observation of nesting suggest that M. scaber is a paired spawner.     

东海产长木叶鲽的年龄与生长

研究了东海产长木叶鲽的年龄与生长等生物学特性。样品取自１９９６ 年４ 月～１９９８ 年１１月,日本以西底拖网渔业在长崎鱼市场上岸的渔获物共２ ５８４ 尾。采用耳石进行年龄鉴定。体长与耳石径的关系,雌雄间有显著性差异。耳石上轮纹每年１～３ 月份形成一次。周龄的推定体长,５＋ 龄以上,雌性偏大     

DNA strand breaks (comet assay) and embryo development effects in grass shrimp (Palaemonetes pugio) embryos after exposure to genotoxicants.

Grass shrimp embryos develop in egg sacs (stages 1-10) attached to the female for 14-20 days after which they 'hatch' from the egg sacs into a swimming zoea stage (stage 11). Until they emerge from the egg sacs, embryos depend on lipids and lipovitellin stored within the egg. The percent of embryos which hatch after exposure to toxicants relative to controls was the basis of an embryo development assay. Exposure of embryos to chromium(III) chloride, sodium chromate, mercuric chloride, and 2-methyl-1,2-naphthoquinone (MNQ) resulted in a reduced hatching rate. In addition to effects on embryo development, DNA strand damage tests were carried out on contaminant-exposed embryos, using the single-cell electrophoresis method often referred to as comet assay. Development of stage 4 embryos was more affected by MNQ exposure than stage 7 embryos. The hatching rates of stages 4 and 7 embryos exposed to MNQ (172 micrograms/l) were 0 and 90%, respectively. DNA strand damage, measured as DNA tail moments, were 3.4 and 4.4, respectively. Thus, exposure of an early embryo stage to MNQ prevented full embryo development while development of later embryo stages was not affected. It may be that the DNA repair systems are more efficient in later embryo stages than in early stages and thus DNA damaged in the early stages affects development.     

Describing gonad development and gametogenesis in southern Africa’s endemic box jellyfish Carybdea branchi (Cubozoa,Carybdeidae)

Information on the reproductive development of species of box jellyfish (class Cubozoa) is poor globally, despite their significance as potent stingers and worrying projections about range expansions. While most species are confined to tropical and subtropical waters, the South African box jellyfish Carybdea branchi is commonly found in the cold-water (especially southern) Benguela ecosystem. Its biology is unknown. Here, we examine reproductive development and describe gametogenesis in this dioecious species. Four and five maturity classes were determined for males and females, respectively, as well as five oocyte developmental stages. Oocyte sizes differed significantly between developmental stages. Mature and immature medusae differed significantly in size, but males and females did not. Individuals matured at approximately 30 mm average bell dimensions. Carybdea branchi displayed gonadal development characteristic of a semelparous organism.