Construction and characterization of a fosmid library for pathogenic bacterium Vibrio anguillarum

Vibrio anguillarum is a common bacterial pathogen in fish. However, little is known about its pathogenic mechanism, in part, because the entire genome has not been completely sequenced. We constructed a fosmid library for V. anguillarum containing 960 clones with an average insert size of 37.7 kb and 8.6-fold genome coverage. We characterized the library by end-sequencing 50 randomly selected clones. This generated 93 sequences with a total length of 57 485 by covering 1.4% of the whole genome. Of these sequences, 58 (62.4%) were homologous to known genes, 30 (32.3%) were genes with hypothetical functions, and the remaining 5 (5.3%) were unknown genes. We demonstrated the utility of this library by PCR screening of 10 genes. This resulted in an average of 6.2 fosmid clones per screening. This fosmid library offers a new tool for gene screening and cloning of V. anguillarum, and for comparative genomic studies among Vibrio species.

Feeding resumption, morphological changes and mortality during starvation in Japanese flounder larvae

Japanese flounder Paralichthys olivaceus larvae established first feeding 3 days after hatching (DAH) at c. 17degreesC. Non-fed fish reached irreversible starvation at age 5 DAH. Non-fed fish showed similar feeding rate and feeding intensity as the fed fish when they were provided with prey before 5 DAH, after which the starved larvae did not feed even when prey became available. None of the six morphological measurements examined (total length, body height, eye height, head height, gut height and myotome height) showed significant differences between the non-fed and fed larvae until 5 DAH. Normal development continued only in the fed group, and the non-fed larvae showed reverse growth or body collapse after 5 DAH. Owing to the shrinkage and collapse at the top of head due to starvation, head height could be a sensitive indicator of starvation in Japanese flounder larvae. In the fed treatments, high mortality occurred from first feeding (3 DAH) to irreversible starvation (5 DAH), accounting for about two-thirds to three-quarters of the overall mortality (46-52%) throughout the experiments. This mortality was not prey density or larval density dependent. Mortality during the same period in the non-fed larvae accounted for about a third of the overall mortality (100%). (C) 2002 The Fisheries Society of the British Isles. Published by Elsevier Science Ltd. All rights reserved.

Effects of photoperiod on growth, mortality and digestive enzymes in miiuy croaker larvae and juveniles

The growth, mortality and digestive enzymes (trypsin, amylase and lipase) in miiuy croaker Miichthys miiuy larvae and juveniles (2-53 dph) were investigated at four photoperiods: 24L:OD), 18L:6D, 12L:12D and OL:24D. Larvae could not feed at OL:24D and did not survive up to 7 dph. In the 24L:OD, 18L:6D, 12L:12D groups, photoperiod had not significant effects on the growth of the rniiuy croaker younger than 20 dph. However, their total length and specific growth rate (SGR) were significantly larger at 18L:6D and 24L:OD than 12L:12D after 20 dph. Photoperiod also affected the mortality of the first feeding larvae (5 dph). being apparently higher in 5 dph larvae at OL:24D (60%) than at other photopenods (20-27%), but no significant differences in mortality were found among other photoperiods. High mortality of the miiuy croaker in 12L:12D, 18L:6D and 24 L:OD groups mainly occurred from 5 (20-27%) to 11 dph (11-16%) and tended to decrease gradually from 15 dph onwards. Digestive enzymes activities in the rniiuy croaker larvae and juveniles had a similar change trend with age at all photoperiods. They underwent drastic changes with age. The specific activity of lipase was significantly higher at 18L:6D and 24L:0D than 12L:12D, but there were no significant differences in specific activities of either trypsin or amylase among photoperiods. With regard to the total length, SGR, survival and digestive enzyme activities, our findings suggested that the optimal light regime for the culture of miiuy croaker during the early life stage was 18L:6D. (C) 2008 Elsevier B.V. All rights reserved.

Necessity of dietary lecithin and eicosapentaenoic acid for growth, survival, stress resistance and lipoprotein formation in gilthead sea bream Sparus aurata

The substitution of dietary docosahexaenoic acid (DHA) with eicosapentaenoic acid (EPA) reduces larval growth in gilthead sea bream. However, the value of EPA when dietary DHA is able to meet the requirements of the larvae has not been sufficiently studied. Dietary phosphoacylgliceride levels also affect fish growth and it has been suggested that they enhance lipid transport in developing larvae. The present experiment was carried out to further study the effect of dietary lecithin and eicosapentaenoic acid on growth, survival, stress resistance,. larval fatty acid composition and lipid transport, when DHA is present in the microdiets of gilthead:sea bream. Eighteen thousand gilt-head sea bream larvae of 4.99+/-0.53 mm total length were fed three microdiets tested by triplicate: a control diet [2% soybean lecithin (SBL) and 2.89% EPA], a low EPA diet,(2% SBL and 1.63% EPA) and a no SBL diet (0% SBL and 2.71% EPA). Handling, temperature and salinity tests determined larval resistance to stress. The results show that when dietary DHA levels are high, but dietary arachidonic acid (ARA) levels are about 0.2%, EPA is necessary to improve larval growth, and survival. Larval EPA content, but not DHA or ARA, was affected by dietary EPA levels. Increased dietary EPA improved larval stress resistance to handling and temperature tests, which could be related to its possible role as a regulator of cortisol production whereas it did not affect stress resistance after salinity shock. Larvae fed the no SBL diet showed a lower lipid content characterized by a low proportion of saturated and monounsaturated fatty acids, together with a significant reduction in the appearance of lipoprotein particles in the lamina propria and in the size of such particles, denoting a critical reduction in dietary lipid transport and utilization, and lower larval growth and survival rates.

Genetic improvement on Chinese shrimp (Fenneropenaeus chinensis): growth and viability performance in F1 hybrids of different populations

Fenneropenaeus chinensis distributed in the Yellow Sea and Bohai Sea of China and the west coast of the Korean Peninsula. Different geographical populations represent potentially different genetic resources. To learn further the characteristics of different geographical population, crosses among two wild and three farmed populations were produced. The two wild populations were from the Yellow Sea and Bohai Sea (WYP), and the west coast of the Korean Peninsula and coast (WKN). The three farmed populations included the offspring of first generation of wild shrimp from coast in Korea (FKN), the Huang Hai (the Yellow Sea in Chinese) No.1 (HH1), and JK98. The phenotypes growth and survival rates of these populations were compared to confirm the feasibility for crossbreeding. The body length (BL), carapace length (CL), carapace width (CW), height of the second and third abdominal segment (HST), width of the second and third abdominal segment (WST), length of the first abdominal segment (LF), length of the last abdominal segment (LL), live body weight (BW), and survival rate were measured. Different combinations were statistically performed with ANOVA and Duncan's Multiple Range Test. The results show that the survival rate of JK98(a (TM) Euro)xWKN(a (TM),) was the highest, followed by WYP(a (TM) Euro)xWKN(a (TM),), FKN(a (TM) Euro)xWYP(a (TM),), FKN(a (TM) Euro)xHH1(a (TM),) and WYP(a (TM) Euro)xFKN(a (TM),); the body weight of FKN(a (TM) Euro)sxHH1(a (TM),) was the highest, followed by FKN(a (TM) Euro)xWYP(a (TM),), WYP(a (TM) Euro)xWKN(a (TM),), WYP(a (TM) Euro)xFKN(a (TM),) and JK98(a (TM) Euro)xWKN(a (TM),); the total length had the same ranking as the body weight. All growth traits in hybrids JK98(a (TM) Euro)xWKN(a (TM),) were the lowest among all combinations. F1 hybrids had significant difference (P < 0.05) in BL, CL, HST, LL, and BW; and insignificant difference (P > 0.05) in other growth traits and survival rate. The results of Duncan's Multiple Range Test are that BL and CL of JK98(a (TM) Euro)xWKN(a (TM),) were significantly different from the other combinations; HST different from the combination of FKN(a (TM) Euro)xWYP(a (TM),), FKN(a (TM) Euro)xHH1(a (TM),) and WYP(a (TM) Euro)xWKN(a (TM),); and BW different from FKN(a (TM) Euro)xWYP(a (TM),) and FKN(a (TM) Euro)xHH1(a (TM),). As a whole, the results indicate that the FKN(a (TM) Euro)xHH1(a (TM),) was the best combination in all growth traits. Therefore, hybridization can introduce the variation to base populations. The systematic selection program based on additive genetic performance may be more effective than crossbreeding.

Separation and determination of flavone and xanthone glycosides in Tibetan folk medicinal species Swertia mussotii and S-franchetiana by capillary electrophoresis

The contents of five pharmacologically active flavone and xanthone glycosides, namely, swertianolin, swertisin, isoorientin, mangiferin, and 7-O-[alpha-L-rhamnopyranosyl-(1 -> 2)-beta-D-xylopyranosyl]-1,8-dihydroxy-3-methoxyxanthone, extracted from Tibetan folk medicinal species Swertia mussotii and S. franchetiana were determined by capillary electrophoresis with diode-array detection. The separation of five components has been optimized with a capillary column with a total length of 48.5 cm and effective length of 40 cm (50 mu m i.d). The influence of the running buffer, the sodium dodecyl sulfonate (SDS) concentration, organic modifier, etc. on the resolution was evaluated. The background electrolyte contained 30 mM borate buffer, 28 mM SDS, 1.0% (v/v) acetonitrile, and was adjusted to pH 9.0 with 0.1 M NaOH. A good baseline resolution was obtained for the separation of five components within 5 min with the working voltage of 24 kV and a column temperature of 25 degrees C. The established method was rapid and reproducible for the separation and determination of five flavone and xanthone glycosides from the extracts of S. mussotii and franchetiana plant samples.

CE determination of 2-(9-carbazole)ethyl chloroformate-labeled oligopeptides

A pre-column derivatization method for sensitive determination of oligopeptides, using the tagging reagent 2-(9-carbazole)ethyl chloroformate (CEOC-Cl) followed by capillary electrophoresis (CE) with diode-array detection, has been developed. Maximum yield close to 100% were observed when a three to fourfold molar excess of reagent was used at pH 9.0-10.0. Excess reagent was extracted with n-hexane-ethyl acetate 9:1-10:1 (v/v); this enabled direct analysis using CE with no significant disturbance from the major fluorescent reagent degradation by-products. The effects on the results of buffer pH and of SDS and organic modifier concentrations were examined. Good baseline resolution in the separation of five CEOC-peptides was achieved with a 48.5-cm total length (effective length 40 cm) 50-mu m inner diameter capillary column.

基于GIS的高速公路养护管理信息系统的设计与开发

China is experiencing a rapid development of highway ever since 1990s. By the end of 2004, the total length of the highway summed up to 33 thousand kilometers, ranking 2n in the world. After the open of highway, the accumulation of time and traffic causes the decrease of its capability. To ensure its good quality, security and operation functions, we should take some reasonable measures to maintain it periodically. At present, a big problem is that the traditional maintain measures can no longer meet the increasing requirements. Due to the characters of highway, the relationship of various maintenance data and geographic positions is even closer than any others. If we wan to improve the quality and efficiency of the maintenance work, particularly when there is need for decision-making, a great number of data that is related to geographic positions are absolutely necessary. Evidently, Geographical Information System (GIS) has incomparably advantages in dealing with these spatial information. As a result, a management system for highway maintenance work based on GIS became inevitable for the development of the maintenance of highway. The purpose of this paper is to establish a management system for highway maintenance work base on Geographical Information System (GIS), Global Positioning System (GPS) and spatial database, to manage all kinds of problems encountered in the work, and to provide support on information and methods. My study mainly includes: (1) Analysis on the current status of the maintenance and management work; overview on the history of domestic and international highway maintenance management systems; identifying the necessity and importance for establishing a management system for highway maintenance work based on GIS. (2) Based on the requirement analysis, I proposed a general design for this management system, and discussed the objective, design principles, framework, systematical structure and function design. (3) Outdoor data collection is not only a prime way to understand the current situation of the road, but also an important method for data update after the system is put into use. This paper also intends to establish a set of plan to collect data efficiently and precisely which is based on GIS and GPS technologies. (4) The maintenance management database is a supporting platform for various maintenance decision-makings. Such decisions need the support of a great amount of data, which would cause other problems, such as the diversity of the data source, difference of data formats. This paper also discussed how to deal with these problems and establish such a database. (5) To propose an approach to assess the condition of pavement, based on GIS and related maintenance models. Among all the maintenance models, the two for assessing and forecasting pavement condition are the most important and mature. This paper also analyzed these two models and introduced them in terms of the integration of models. (6) This paper took the Guangshen Highway for example, explaining how to realize a GIS for management of highway maintenance work.

Discussions on slip length measurements by microPIV/PTV in microchannels

Fluid transportation in microfluidic system could be benefit from the slip on solid-liquid interface. Slip length on many kinds of hydrophilic/hydrophobic surfaces have been measured recently. The two common-used experimental methods for boundary slip measurement include: (1) surface force measurement, such as surface force apparatus (SFA), atom force microscope (AFM), and (2) velocity measurement, like microPIV/PTV (Particle image velocimetry / Particle tracking velocimetry), total internal reflection velocimetry (TIRV). However, the measured results are rather scattered, larger measured slip lengths were reported by microPIV/PTV experiments. In this paper, we will investigate the deviations of the measured slip length on smooth hydrophilic surface. After measuring detailed velocity profiles very close to hydrophilic glass wall, we give a discussion on the effects influencing the slip measurements.

Influence of Coulomb potential for photoionization of H atoms in an elliptically polarized laser field: Velocity gauge versus length gauge

We theoretically study the influence of Coulomb potential for photoionization of hydrogen atoms in an intense laser field with elliptical polarization. The total ionization rates, photoelectron energy spectra, and photoelectron angular distributions are calculated with the Coulomb-Volkov wave functions in the velocity gauge and compared with those calculated in the length gauge as well as those calculated with the Volkov wave functions. By comparing the results obtained by the Coulomb-Volkov and Volkov wave functions, we find that for linear polarization the influence of Coulomb potential is obvious for low-energy photoelectrons, and as the photoelectron energy and/or the laser intensity increase, its influence becomes smaller. This trend, however, is not so clear for the case of elliptical polarization. We also find that the twofold symmetry in the photoelectron angular distributions for elliptical polarization is caused by the cooperation of Coulomb potential and interference of multiple transition channels. About the gauge issue, we show that the difference in the photoelectron angular distributions obtained by the velocity and length gauges becomes rather obvious for elliptical polarization, while the difference is generally smaller for linear polarization.

Amplified fragment length polymorphism analysis to identify the genetic structure of the Gymnocypris przewalskii (Kessler, 1876) population from the Qinghai Basin, China

Amplified fragment length polymorphism (AFLP) was used to analyse the genetic structure of 45 individuals of Gymnocypris przewalskii (Kessler, 1876), an endangered and state-protected rare fish species, from three areas [the Heima (HM), Buha (BH) and Shaliu rivers (SL), all draining into Qinghai Lake]. A total of 563 polymorphic loci were detected. The HM, BH and SL populations have 435, 433 and 391 loci, respectively (Zhu and Wu, 1975), which account for 77.26%, 76.91% and 69.45% of the total number of polymorphic loci of each population, respectively. The Nei indices of genetic diversities (H) of the three populations were calculated to be 0.2869 (HM), 0.2884 (BH) and 0.2663 (SL), respectively. Their Shannon informative indices are 0.4244, 0.4251 and 0.3915, respectively. Research results show that the mean genetic distance between HM and BH is the smallest (0.0511), between BH and SL is the second shortest (0.0608), and between HM and SL is the largest (0.0713), with the mean genetic distance among the three populations being over 0.05. Data mentioned above indicate that the three populations have a certain genetic differentiation. The total genetic diversity (H-t = 0.3045) and the mean value of genetic diversity within the population (H-s = 0.2786) indicate that the variations have mainly come from within the population.

A lattice dynamical treatment for the total potential energy of single-walled carbon nanotubes and its applications: relaxed equilibrium structure, elastic properties, and vibrational modes of ultra-narrow tubes

In this paper, we propose a lattice dynamic treatment for the total potential energy of single-walled carbon nanotubes (SWCNTs) which is, apart from a parameter for the nonlinear effects, extracted from the vibrational energy of the planar graphene sheet. The energetics, elasticity and lattice dynamics are treated in terms of the same set of force constants, independently of the tube structures. Based upon this proposal, we have investigated systematically the relaxed lattice configuration for narrow SWCNTs, the strain energy, the Young's modulus and Poisson ratio, and the lattice vibrational properties with respect to the relaxed equilibrium tubule structure. Our calculated results for various physical quantities are nicely in consistency with existing experimental measurements. In particular, we verified that the relaxation effect makes the bond length longer and the frequencies of various optical vibrational modes softer. Our calculation provides evidence that the Young's modulus of an armchair tube exceeds that of the planar graphene sheet, and that the large diameter limits of the Young's modulus and Poisson ratio are in agreement with the experimental values of graphite; the calculated radial breathing modes for ultra-narrow tubes with diameters ranging between 2 and 5 angstrom coincide with the experimental results and the existing ab initio calculations with satisfaction. For narrow tubes with a diameter of 20 angstrom, the calculated frequencies of optical modes in the tubule's tangential plane, as well as those of radial breathing modes, are also in good agreement with the experimental measurements. In addition, our calculation shows that various physical quantities of relaxed SWCNTs can actually be expanded in terms of the chiral angle defined for the corresponding ideal SWCNTs.

Growth and characterization of 0.8-mu m gate length AlGaN/GaN HEMTs on sapphire substrates

AlGaN/GaN high electron mobility transistor (HEMT) structures were grown on 2 inch sapphire substrates by MOCVD, and 0.8-mu m gate length devices were fabricated and measured. It is shown by resistance mapping that the HEMT structures have an average sheet resistance of approximately 380 Omega/sq with a uniformity of more than 96%. The 1-mm gate width devices using the materials yielded a pulsed drain current of 784 mA/mm at V-gs=0.5 V and V-ds=7 V with an extrinsic transconductance of 200 mS/mm. A 20-GHz unity current gain cutoff frequency (f(T)) and a 28-GHz maximum oscillation frequency (f(max)) were obtained. The device with a 0.6-mm gate width yielded a total output power of 2.0 W/mm (power density of 3.33 W/mm) with 41% power added efficiency (PAE) at 4 GHz.

SOI optical switch matrix integrated with spot size converter (SSC) and total internal reflection (TIR) mirrors

SOI (Silicon on Insulator) based photonic devices has attracted more and more attention in the recent years. Integration of SOI optical switch matrix with isolating grooves, total internal reflection (TIR) mirrors and spot size converter (SSC) was studied. A folding re-arrangeable non-blocking 4x4 optical switch matrix and a blocking 16x16 matrix with TIR mirrors and SSC were fabricated on SOI wafer. The performaces, including extinction ratio and the crosstalk, are better than before. The insertion loss and the polarization dependent loss (PDL) at 1.55 mu m increase slightly with longer device length, more bend and intersecting waveguides. The insertion losses decrease 2 similar to 3 dB when anti-reflection films are added in the ends of the devices. The rise and fall times of the devices are 2.1 mu s and 2.3 mu s, respectively.

QTL Mapping for Frond Length and Width in Laminaria japonica Aresch (Laminarales, Phaeophyta) Using AFLP and SSR Markers

In Laminaria japonica Aresch breeding practice, two quantitative traits, frond length (FL) and frond width (FW), are the most important phenotypic selection index. In order to increase the breeding efficiency by integrating phenotypic selection and marker-assisted selection, the first set of QTL controlling the two traits were determined in F-2 family using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Two prominent L. japonicas inbred lines, one with "broad and thin blade" characteristics and another with "long and narrow blade" characteristics, were applied in the hybridization to yield the F-2 mapping population with 92 individuals. A total of 287 AFLP markers and 11 SSR markers were used to construct a L. japonica genetic map. The yielded map was consisted of 28 linkage groups (LG) named LG1 to LG28, spanning 1,811.1 cM with an average interval of 6.7 cM and covering the 82.8% of the estimated genome 2,186.7 cM. While three genome-wide significant QTL were detected on LG1 (two QTL) and LG4 for "FL," explaining in total 42.36% of the phenotypic variance, two QTL were identified on LG3 and LG5 for the trait "FW," accounting for the total of 36.39% of the phenotypic variance. The gene action of these QTL was additive and partially dominant. The yielded linkage map and the detected QTL can provide a tool for further genetic analysis of two traits and be potential for maker-assisted selection in L. japonica breeding.