Isolation and characterization of 49 polymorphic microsatellite loci in the ayu,Plecoglossus altivelis

A total of 49 novel polymorphic microsatellite markers in the ayu fish, Plecoglossus altivelis, were successfully isolated and characterized. The number of alleles per locus, observed heterozygosity, and expected heterozygosity ranged from two to 25 (11.3 on average), from 0.125 to 1.000 (0.648 on average), and from 0.123 to 0.957 (0.751 on average), respectively. These microsatellite markers will be useful for both studying population structure and identifying quantitative trait loci that control evolutionarily and economically important traits of ayu.     

Utilization of α-starch in ayu, Plecoglossus altivelis, relating to growth and body composition

To examine the possibility of dietary α‐starch in reducing feed costs in a practical diet, α‐starch was supplemented at 10, 20, 30 and 40% in a composed diet having the same protein level. The four diets were fed to ayu, Plecoglossus altivelis (initial weight 9.1 g) for 43 days. Growth and feed efficiency increased with the supplement, with values highest in the 30–40%α‐starch diet. The level of dietary α‐starch did not affect the proximate muscle composition; although the hepatosomatic index was not affected, liver glycogen increased with increasing dietary α‐starch. The dietary α‐starch did not influence evacuation time from the gut, and was well digested through passage in the gut, mainly between the stomach and the anterior part of the intestine. Ayu have an ability to adapt their metabolism to high dietary α‐starch, and can digest 40% or more in a composed diet. Although the muscle lipid content did not change, the fatty acid composition was influenced by dietary starch. With the elevation of dietary starch, a decrease of C18:2n‐6 and an increase of C22:6n‐3 occurred. These results indicate that at least 40%α‐starch can be used in practical diets for ayu.     

Competitive influence of isaza,Chaenogobius isaza,on ayu,Plecoglossus altivelis,in Lake Biwa

1. Competitive influence of the isaza population on the ayu population was studied in Lake Biwa.
2. The results of comparison of ecology of the two species suggest that interaction of ayu and isaza possibly occurs in cold season when there is no other important competitor.
3. The significance of interspecific competition between them is evaluated as a factor controlling the rate of recruitment of juvenile through analysis of the Pre-war catch statistics.
4. Presence of the compensation between the juvenile stage and the adult one relieve the ayu population of influence of interspecific competition which they will suffer in the stage of juvenile.
5. Competitive processes which will be responsible for increase of mortality and reduction of individual growth are discussed.

     

Pattern of synaptic connections in the pineal organ of the ayu,Plecoglossus altivelis (Teleostei)

Summary Synaptic connections were studied by means of electron microscopy in the sensory pineal organ of the ayu, Plecoglossus altivelis, a highly photosensitive teleost species. Three types of specific contacts were observed in the pineal end-vesicle: 1) symmetrically organized gap junctions between the basal processes of adjacent photoreceptor cells; 2) sensory synapses endowed with synaptic ribbons, formed by basal processes of photoreceptor cells and dendrites of pineal neurons; 3) conventional synapses between pineal neurons, containing both clear and dense-core vesicles at the presynaptic site. Based on these findings, the following interpretations are given: (i) The gap junctions may be involved in an enhancement of electric communication and signal encoding between pineal photoreceptor cells. (ii) The sensory synapses transmit photic signals from the photoreceptor cells to pineal nerve cells. (iii) The conventional synapses are assumed to be involved in a lateral interaction and/or summation of information in the sensory pineal organ. A concept of synaptic relationships among the sensory and neuronal elements in the pineal organ of the ayu is presented.Fellow of the Alexander von Humboldt Foundation, Federal Republic of Germany     

香鱼hepcidin基因的克隆、序列分析及组织表达特征

Hepcidin是一类富含半胱氨酸的抗菌肽,在鱼类非特异性免疫中起重要作用,具有调节铁代谢的功能。该研究克隆了香鱼hepcidin基因cDNA序列,全长763个核苷酸,包含一个完整的开放阅读框,推测编码一个由85个氨基酸组成的相对分子质量为9.7k的多肽,N端24个氨基酸是信号肽序列。香鱼hepcidin的成熟肽序列由25个氨基酸组成,含有8个半胱氨酸,可形成4个分子内二硫键结构。系统进化树分析表明,hepcidin的物种进化关系与目前接受的物种分类关系基本一致,香鱼hepcidin位于鱼类hepcidin簇中,与大西洋鲑hepcidin的亲缘关系最近,达60%。香鱼hepcidin在肝脏中表达量最大,在脾、肾、心脏和肌肉中也有表达。实时荧光定量PCR(qRT-PCR)结果表明,鳗利斯顿氏菌(Listonella anguillarum)感染以后香鱼肝组织中hepcidin基因mRNA表达量显著增加,在12h增加了8.26倍。hepcidin基因可能在香鱼抗外界病原物感染过程中起重要作用。     

宁海地区香鱼弧菌病病原菌鉴定

摘要：【目的】香鱼弧菌病对中国沿海地区的香鱼养殖业造成了巨大的危害,然而,病原不明导致了防治上的许多问题。本文鉴定了引起宁海地区香鱼爆发性弧菌病的病原。【方法】采用TCBS平板分离优势菌;采用回归感染试验确认病原菌,采用改进的寇氏法计算LD50;采用形态学观察、生理生化特征测定、细菌特异性引物PCR扩增检测及细菌16S rRNA和金属蛋白酶（MP）基因序列分析鉴定细菌;采用药敏实验测定它对部分抗生素的敏感性。【结果】分离并鉴定优势菌株ayu-H080701为宁海地区香鱼弧菌病的病原菌,它对香鱼的半致死量为1.2×104 CFU。形态学观察和生理生化特征测定表明,ayu-H080701与鳗利斯顿氏菌最为接近。PCR扩增检测表明,细菌16S rRNA 基因通用引物和鳗利斯顿氏菌MP基因特异引物均能扩增到预期大小的特异性条带。ayu-H080701与鳗利斯顿氏菌16S rRNA基因核苷酸序列同源性最高,为99.4%～99.5%,与同属的海弧菌和美人鱼发光杆菌分别为94.3%和91.9%;ayu-H080701与鳗利斯顿氏菌MP氨基酸序列同源性高达97.6%～98.8 %,与其它弧菌科成员则低于75.6 %,系统进化树分析也揭示ayu-H080701与鳗利斯顿氏菌进化相关性最高。【结论】引起宁海地区香鱼弧菌病的菌株ayu-H080701被鉴定为鳗利斯顿氏菌。     

Existence of an extra-retinal and extra-pineal photoreceptive organ that regulates photoperiodism in gonadal development of an Osmerid teleost, ayu (Plecoglossus altivelis)

Ayu (Plecoglossus altivelis) is an Osmerid teleost whose gonadal development exhibits clear photoperiodism: it is stimulated and prevented under short and long photoperiod, respectively. However, the photoreceptor organ involved in this process remains to be identified. In the present study, we examined whether gonads of ophthalmectomized (Ex) and pinealectomized (Px) ayu respond to short photoperiod to test whether photoreceptors other than lateral eyes and pineal complex are involved in the photoperiodic response of gonadal development. Gonadosomatic index (GSI) and plasma levels of sex steroids (testosterone and 11-ketotestosterone for males and testosterone and estradiol 17-beta for females) were significantly increased in the Ex+Px ayu kept under short photoperiod in both males and females as compared with the initial control. On the other hand, there were no significant increases in GSI and sex steroids in the Ex+Px ayu kept under long photoperiod. Histological observation of gonads in the Ex+Px ayu revealed that oocytes undergoing final maturation in females and proliferation of germ cells in males were observed under short photoperiod but not under long photoperiod. These results indicate that extra-retinal and extra-pineal photoreceptive organ regulates photoperiodic gonadal development in this species.     

The use of warmed water treatment to induce protective immunity against the bacterial cold-water disease pathogen Flavobacterium psychrophilum in ayu (Plecoglossus altivelis)

We investigated the induction of protective immunity against bacterial cold-water disease (BCWD) caused by Flavobacterium psychrophilum by warmed water treatment in ayu (Plecoglossus altivelis). Fish were immersed in a live bacterial suspension (10⁷ CFU mL⁻¹) for 30 min and placed in 700 L concrete tanks. The 28 °C warmed water treatment lasted 3 days and began 1, 6, and 24 h after immersion in the live bacterial suspension. A naïve control fish group was immersed in a sterilized modified Cytophaga (MCY) broth instead of the bacterial suspension. Fourteen days after the immersion, agglutination antibody titers against F. psychrophilum were measured by using micro-titer methods. Fish were then exposed to a bacterial bath to infect them with live F. psychrophilum, and cumulative mortality was monitored. Fish treated with warmed water at 1, 6, and 24 h after immersion in the live bacterial suspension had cumulative mortalities of 36%, 30%, and 18%, respectively, all of which were significantly lower than the cumulative mortality of the naïve control fish (90%). Treated fish also showed high antibody titers against F. psychrophilum in agglutination tests. These results demonstrate that warmed water treatment could not only cure BCWD but also immunize the fish against the causative agent F. psychrophilum.     

The establishment of a library screening method based on yeast two-hybrid system and its use to determine the potential interactions of liver proteins in ayu, Plecoglossus altivelis

Knowledge of specific protein–protein interaction (PPI) is an important component in understanding biological processes and regulatory mechanisms. A library to library screening method (LLS) was established based on yeast two-hybrid (YTH) system in this research, and applied to study the PPIs in ayu liver. In total, 23 out of 55 interaction pairs were found positive through phenotypic identification, with a positive rate of 41.8%. Of the 11 unique PPIs, 9 interactions including FGB/FGG, CaM/Spna2, C9/Apo-AI-1, α₂M/Ft, RPL10/RPL5, C8α/C9, FGG/Apo-AI-1, LECT2/Tf, and Apo-AI-2/C9 were previously reported. The other two PPIs including FGG/CLR and Wap65/C3 are novel, and in vitro co-immunoprecipitation (co-IP) experiments further confirmed these interactions. FGG/CLR interaction might play a role in regulating the inflammatory response. The interaction between Wap65 and C3 hints that Wap65 might function through the complement activation pathways when microbial infection occurs.     

池塘养殖三疣梭子蟹雄体生长和性腺发育规律

池塘养殖是三疣梭子蟹(Portunus trituberculatus)的主要养殖方式,但关于池塘养殖三疣梭子蟹雄体的生长和性腺发育规律尚不清楚。本研究通过定点连续采样,首先系统研究池塘养殖过程中三疣梭子蟹雄体的生长和成熟雄体生殖系统的外观及组成的变化,其次研究池塘养殖三疣梭子蟹雄体性腺发育过程中性腺各部分组织学特征及其指数的变化,以及不同月份雄体性腺发育的情况和比例。结果表明:(1)三疣梭子蟹雄体的体重和壳宽增长率及特定增长率均在8月份最高,且体重与壳宽呈显著正相关关系。(2)三疣梭子蟹成熟雄体的生殖系统由体外(阴茎、交接器)和体内(精巢、输精管)两部分构成。(3)根据精巢和输精管的外观、组织学特征及其指数的变化,可将三疣梭子蟹雄体性腺发育分为3期。精子发生期,即精巢内主要细胞类型是精原细胞和初级精母细胞,精巢指数为0.07%～0.16%,此时输精管呈透明状,肉眼很难发现;精荚形成期,精巢内主要细胞类型为次级精母细胞和精子细胞,精巢指数为0.10%～0.51%,输精管内有大量精荚和分泌物,输精管指数为0.01%～0.41%;成熟期,精巢内主要细胞类型是精子细胞和成熟精子,精巢指数为0.10%～0.41%,输精管进一步膨大,输精管指数为0.20%～0.65%。(4)在雄体性腺发育期间,性腺指数和输精管指数呈显著上升趋势,而精巢指数呈现先上升后下降的趋势,且统计分析发现性腺指数与肝胰腺指数无明显的相关性。(5)池塘养殖雄体性腺发育不同步,7月份主要处于精子发生期,8月底,有76%的雄体性腺发育达到精荚形成期,至9月底,达到成熟期雄体的比例为47%,此后雄体性腺发育趋于成熟,10月中下旬至12月份池塘养殖雄体均主要处于成熟期。     

Feeding and growth of larval herring,Clupea harengus,in relation to density of copepod nauplii

Synopsis Feeding and growth rates of 1–3 wk old herring larvae from four different stocks were compared in laboratory experiments (8°C). For most of the larval groups, feeding rate was saturated at nauplii (Acartia tonsa, nauplii stages 3–5) densities over 301^–1 (5 g d.w. 1^–1). Specific growth rate increased asymptotically with nauplii density, and reached about 6% d^–1 at densities over 120 nauplii 1^–1. The growth rates attained in the laboratory were similar to field measured growth rates of similarly aged herring larvae at comparable food densities. Since food particles were homogenously distributed in the laboratory tanks, patches of dense plankton concentrations are, thus, apparently not necessary for larval growth and survival in the sea. Growth efficiency differed between larval groups, with large sized larvae being the most efficient in transforming ingested matter into growth. The difference probably relates to different sizes rather than to the different geographical origins of the larvae.     

Mosquito Control Evaluations of an Insect Growth Regulator, Pyriproxyfen against Culex pipiens pattens (Diptera, Culicidae) Larvae in Marsh Area, Korea

ABSTRACT Mosquito control evaluations of the granular formulation of 0.5% pyriproxyfen were undertaken in marshes and a pond in Busan during the period of July through September, 2001. In marsh condition where Culex pipiens pattens were predominantly breeding, the treatment of pyriproxyfen produced over an average of 95% mosquito larval reduction at a concentration of 0.05 mg/1 for the first 4 weeks in spite of heavy precipitation (total 274.4 mm) during the period. Reduction rates of 72.6% and 32.3% appeared in the 5th and 7th weeks probably due to heavy input of sewage and flood. However, the reduction rate increased again at the 8th week, showing a 94.5% reduction, probably because of the slow releasing of pyriproxyfen granular formulation. In the second treatment of pyriproxyfen, complete mortality was obtained at the 1st and 3rd weeks. Also, a satisfactory level of Cx. pipiens larval reduction of 96.6% was observed at the 4th week. In a pond assessment, the treatment of pyriproxyfen at a concentration of 0.05 mg/1 produced a complete reduction of Cx. pipiens pattens from the 1st week through the 4th week after treatment. In the second treatment, 100.0% reduction was obtained at the 1st and 2nd weeks in the same sites. The treatment resulted in 84.4% and 97.9% reduction of mosquitoes at the 3rd and 4th weeks, respectively.     

Ecological studies on the population of isaza,Chaenogobius Isaza Tanaka,in Lake Biwa,with special reference to the effects of population density upon its growth

1. In order to get the knowledge on the age composition of “isaza” population in Lake Biwa and the effect of population density on growth, monthly distribution of mean body length and mean body weight has been analyzed on the basis of monthly haul by “isazabiki” trawl during 1949 to 1953 and also 1960 to 1965.
2. There is no apparent sex difference in the growth in the first and second years of life.
3. “Isaza” population is composed of two age groups, age 0 and 1 groups (1+fish), the latter occupying by far the greater part in commercial catch.
4. During the growth season fishes of both ages feed mainly on zooplankton, though in winter frequently take chironomid larvae, gammaridae and others in volume.
5. The growth season falls in the period from April to October in both age groups.
6. A considerable yearly variation occurring in growth is in close connection with the fluctuation of population density of all ages.
7. The influence affected by the density of age 1 group is larger than that by age 0 group.

     

Influence of parental density on the distribution pattern of eggs in the common cabbage butterfly,Pieris rapae crucivora

To investigate the relation between the distribution pattern of eggs and the parental density in the common cabbage butterfly, Pieris rapae crucivora, the countings of egg number per plant were made on both cabbage plants cultivated in the farm and planted in the net house in which the female butterflies were released at various densities. The frequency distribution of eggs fits well to the negative binomial excepting the cases where they agree withPoisson series, and the degree of aggregation expressed as the reciprocal of the parameter, 1/k, tends to decrease as the egg or parental density increases. At the same parental density, however, the distribution of eggs can be described by the negative binomial with a common parameter, k_c, regardless of the difference in the density of laid eggs. In the case where a single female butterfly lays eggs, the spatial pattern of egg distribution is always lean, while its frequencies conform toPoisson or the negative binomial series. This lean changes toward patchy with increasing the parental density. From these results, it is concluded that the degree of aggregation in the distribution of eggs decreases with the increase of the parental density.     

The fitting of the logistic equation to the rate of increase of population density

Though there are many problems on the usefulness of the logistic curve, it may be necessary to examine before discussing these problems whether or not the actual data fit to the theoretical values. It has been clarified in this paper that the relation between the population density and its rate of increase per individual described by the differential equation (1) is represented by a straight line on a finite difference diagram on which N_i+1−N_i/N_i values are plotted against N_i+1. Utilizing this linear relation we may examine the fittness of the logistic curve to the actual data and when it is fitted we may estimate the parameters of the logistic equation by (5) and (6). The result of the application of this method to the experimental populations of azuki bean weevil indicates that the relation between parent and progeny densities fits well to the logistic type as has been proved byFujita andUtida (1953) who utilized the linear reltion between 1/R+2σ and parent density where R is the apparent rate of reproduction and σ is a constant dependent primarily upon the length of adult life (0≦σ≦1).     

池塘养殖中华绒螯蟹二龄雄体生长、性腺发育和第二性征的变化研究

为进一步了解中华绒螯蟹(Eriocheir sinensis)2龄雄体在池塘养殖条件下的生长发育规律,本研究采用养殖实验、活体解剖和显微观察方法,首先研究了池塘养殖过程中二龄雄体的生长、性腺指数(GSI)、肝胰腺指数(HSI)和第二性征(主要包括大螯绒毛长度及其覆盖比例)的变化情况,并探讨了这些指标间的相关性,其次,对成熟雄体的生殖系统外观和组成进行了拍照和描述.结果显示:(1)池塘养殖条件下中华绒螯蟹2龄雄体的体重、甲壳长、甲壳宽和体厚增长主要发生在3～8月,其中4～5月间的增重率和特定增长率最高,9～12月间的体重和形态学参数增长均不显著;肥满度总体上呈先下降后上升的趋势.(2)2龄雄体池塘养殖早期性腺发育较慢,3～5月肉眼很难发现,6月份生精区已经清晰可见,7～11月份性腺指数显著增加,7月份部分个体储精囊中已有精荚,8月份副性腺体积开始迅速增加,整体上,肝胰腺指数(HSI)和性腺指数(GSI)呈显著负相关,而性腺指数(GSI)与生殖蜕壳前的雄体体重及月份均呈显著正相关.(3)2龄雄体的大螯绒毛长度及其内侧绒毛覆盖比例在4～8月呈显著增长趋势,其余月份增加不显著,绒毛长度与覆盖比例、绒毛长度与体重、绒毛覆盖比例与体重、绒毛长度与性腺指数、绒毛覆盖比例与性腺指数都呈显著正相关.(4)2龄成熟雄体的生殖系统由体内和体外两大部分构成,体内部分主要包括精巢、输精管(分为前、中、后三个部分)、副性腺和射精管,输精管后端储存着大量精荚,体外部分主要由阴茎和交接器组成.综上,池塘养殖中华绒螯蟹2龄雄体的生长、性腺发育与其第二性征关系密切,雄体生殖系统组成复杂,今后需要深入研究雄体的性腺发育规律和分期.