Construction and characterization of a bacterial artificial chromosome library of marine macroalga Porphyra yezoensis (Rhodophyta)

A large-DNA-fragment library is necessary for research into the Porphyra genome. In this study, a bacterial artificial chromosome (BAC) library of Porphyra yezoensis was constructed and characterized. The library contains 54,144 BAC clones with an average insert size of about 65 kb and fewer than 0.7% of clones without large inserts. Therefore, its capacity is more than 6.6 P. yezoensis genome equivalents, and the probability of recovering any nuclear DNA sequence from the library is higher than 99%. The library shows good fidelity and stability. A putative trehalose-6-phosphate synthase (TPS) gene was successfully screened out from the library. The above results show that the library is useful for gene cloning and genomic research in P. yezoensis.

Diet-induced changes in lipid and fatty acid composition of Artemia salina

The influence of diet on lipid and fatty acid composition of the brine shrimp Artemia salina nauplii was investigated. Various diets with different lipid composition and fatty acid profiles were fed to nauplii for 2 weeks. The lipid composition of microalgal diets, Isochrysis galbana, Phaeodactylum tricornutum and Nannochloropsis oculata and baker's yeast was analyzed. Newly hatched nauplii were examined before the feeding experiment. It was shown that Artemia was able to incorporate and selectively concentrate some dietary lipids. Depot lipids were more sensitive to changes in the dietary lipid composition than the main structural lipids, polar lipids and sterols. Variations in the content of the lipid classes correlated with stage of development of the animal. The fatty acid composition of the animal varied with that of diet. The concentrations of saturated fatty acids were apparently supported in the nauplii by biosynthesis de novo. The acid 16:1(n-7) originated from the food. The concentration range of n-6 polyunsaturated fatty acids (PUFAs) remained constant through the accumulation from the diet. The proportion of n-3 PUFAs varied with their level in the diet. The dynamics of alteration of 20:5(n-3) content in Artemia fed on Isochrysis, which is poor in this acid, suggested a limited capacity for elongation and desaturation of 18:3(n-3) to 20:5(n-3). None of the diets provided dietary input of 22:6(n-3). (C) 1998 Elsevier Science Inc. All rights reserved.

Size and time of day affect the response of postlarvae and early juvenile grooved tiger prawns Penaeus semisulcatus De Haan (Decapoda:Penaeidae) to natural and artificial seagrass in the laboratory

Juvenile tiger prawns (Penaeus semisulcatus De Haan and P. esculentus Haswell) show a strong association with vegetated habitats and are rarely caught on non-vegetated areas. This pattern of distribution may be caused by postlarvae selecting vegetation when they settle, or to differences in post-settlement mortality in different habitats. In this study, we examined whether the postlarvae and early juvenile stages of P. semisulcatus would distinguish between seagrass (Zostera capricorni Aschers) without epiphytes, artificial seagrass and bare substratum in the laboratory. The responses of prawns reared from the egg to different stages of postlarval and juvenile development were tested to determine whether, and when, each size class showed a response to a particular habitat. Five size classes of postlarvae (average carapace lengths [CL] of 1.2, 1.4, 1.6, 1.7 and 2.0 mm) were offered a choice between Z. capricorni and bare sand. Small size classes of postlarvae either did not respond to Z. capricorni (1.2 and 1.6 mm CL), or were more abundant on bare substratum than Z. capricorni. In contrast, the largest size classes of postlarvae (1.7 and 2.0 mm CL) were more abundant on Z. capricorni during the day but not at night. The behaviour of postlarvae changed markedly at a size of 1.7 mm CL (22 days from the first nauplius): smaller postlarvae frequently swam in the water column; 1.7 and 2.0 mm CL postlarvae spent much more. time resting on the substrate and perched on seagrass leaves. This size at which postlarvae first respond to seagrass during the day, and show mainly benthic behaviour, is similar to the size at which they are found on shallow seagrass beds in northern Australia. Large postlarvae (2.7 mm CL) and juveniles (4.1 mm CL) both were more abundant on artificial seagrass than bare sand during the day but not at night, indicating that they respond to structured habitats. When large postlarvae (2.4 mm CL) and juveniles (3.5 mm CL) were offered a choice between Z. capricorni without epiphytes and artificial seagrass, they were more abundant on the Z. capricorni, which suggests that chemical cues from seagrass may explain some of the responses of P. semisulcatus to seagrass. (C) 1997 Elsevier Science B.V.

Construction and characterization of two bacterial artificial chromosome libraries of Zhikong scallop, Chlamys farreri Jones et Preston, and identification of BAC clones containing the genes involved in its innate immune system

Large-insert bacterial artificial chromosome (BAC) libraries are necessary for advanced genetics and genomics research. To facilitate gene cloning and characterization, genome analysis, and physical mapping of scallop, two BAC libraries were constructed from nuclear DNA of Zhikong scallop, Chlamys farreri Jones et Preston. The libraries were constructed in the BamHI and MboI sites of the vector pECBAC1, respectively. The BamHI library consists of 73,728 clones, and approximately 99% of the clones contain scallop nuclear DNA inserts with an average size of 110 kb, covering 8.0x haploid genome equivalents. Similarly, the MboI library consists of 7680 clones, with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1x. The combined libraries collectively contain a total of 81,408 BAC clones arrayed in 212 384-well microtiter plates, representing 9.1x haploid genome equivalents and having a probability of greater than 99% of discovering at least one positive clone with a single-copy sequence. High-density clone filters prepared from a subset of the two libraries were screened with nine pairs of Overgos designed from the cDNA or DNA sequences of six genes involved in the innate immune system of mollusks. Positive clones were identified for every gene, with an average of 5.3 BAC clones per gene probe. These results suggest that the two scallop BAC libraries provide useful tools for gene cloning, genome physical mapping, and large-scale sequencing in the species.

Construction of bacterial artificial chromosome libraries for Zhikong Scallop Chlamys farreri

Two Large-insert genomic bacterial artificial chromosome (BAC) libraries of Zhikong scallop Chlamys farreri were constructed to promote our genetic and genomic research. High-quality megabase-sized DNA was isolated from the adductor muscle of the scallop and partially digested by BamH I and Mbo I, respectively. The BamH I library consisted of 53 760 clones while the Mbo I library consisted of 7 680clones. Approximately 96 % of the clones in BamH I library contained nuclear DNA inserts in average size of 100 kb, providing a coverage of 5.3 haploid genome equivalents. Similarly, the Mbo I library with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1 haploid genome equivalents.

Settling abalone veliger larvae in a free-swimming microalgal culture

In the current abalone hatchery in China, insufficient diatoms on vertically placed corrugated pvc plates at later stage often could not support the growth of postlarvae up to the stage that they can feed on live macroalgae. As a result, stripping the spats (35 mm) off by anaesthetization and switching the diet from live diatoms to artificial powdered diet in combination has to be performed in most of the abalone farms. This manipulation normally leads to more than 50% mortality. Here we report the direct use of the unicellular green alga Platymonas helgolandica Kylin var. tsingtaoensis as a potential alga to be used to settle the veliger larvae of the Pacific abalone Haliotis discus hannai and to feed the postlarvae. Settlement rate of 2-day-old veliger larvae in mono culture of P helgolandica could be as high as 92% ( +/- 4.2%) on day 10 in small scale trials, higher than that in the selected benthic diatom strain (53.6% +/- 12.7%) when settled in the water in which bacteria propagation was controlled by treatment of 2 ppm of benzylpenicillinum calcium and streptomycin sulfate. Postlarvae fed solely on P. helgolandica or the selected benthic diatom Navicula-2005-A grew at rates of 40.1 ( +/- 1.9) and 45.8 (+/- 13.4) mu m day(-1), respectively, when raised at 22 degrees C until day 50 postfertilization. P. helgolandica was shown to have distinct diurnal settling rhythm characterized with a peak of settled cells in the middle of the night for cell division and a peak of free-swimming cells in the middle of the day. High density of attached P. helgolandica cells on the inner surface of the culture facility in the night fits the nocturnal feeding behavior of the abalone spats. Judged by the promising larvae settling rate, growth and survival rates of the postlarvae fed with this alga, the free-swimming micro-green alga P. helgolandica constitutes a potential species for settling the veliger larvae and for supporting the growth of postlarvae as well. (c) 2006 Published by Elsevier B.V.

C, N and P regeneration by a detritivorous fish, Liza haematocheila T. & S.: effects of temperature, diet and body size

Detritus, as a nutrients reservoir, affects the trophic structure and dynamics of communities and supports a greater diversity of species and longer food chains. Detritivorous fish is an important organism to regenerate the nutrients from sediments. Despite the numerous studies on the nutrients cycle in fish, only a few attempts have been made to quantify the regenerating ability. In the present study, we chose the common detritivorous fish redeye mullet as the research object. Redeye mullet is also a common poly-culture fish in China. Diet, including a commercial diet mostly used in aquaculture and a home-made diet with contents close to detritus, was used and considered as a fixed factor. Temperature was also considered as a fixed factor as much research has shown that temperature has significant effects on fish metabolism. Moreover, body size was regarded as a covariate under analysis of covariance. Three key nutrients, namely carbon, nitrogen and phosphorus, were used to measure the nutrient-regenerating ability of redeye mullet under laboratory conditions. The results showed that the nutrient regeneration in percent of the consumption decreased with increasing temperature. Carbon and nitrogen regeneration of redeye mullet fed on commercial diet was lower than those of the home-made diet group, while the opposite was found for phosphorus. In each group, the amount of regenerated nutrients increased linearly with body size. Fed on the home-made diet, 5-g fish at 25 degrees C can regenerate 210.822 mg C, 37.533 mg N and 0.727 mg P per day.

Effects of diet and body size on phosphorus utilization of Liza haematocheila T. & S.

A 21-d laboratory experiment was conducted to study, the phosphorus (P) utilization of two different diets by redlip mullet Liza haematocheila T. & S. Sand-filtered water in salinity 30 and temperature 25 degrees C was used. Twenty-nine fish individuals were divided into three groups: 11 to group 1 (G1) fed on diet 1, 11 to group 2 (G2) fed on diet 2, and 7 to contrast group. Diet 1 was a commercial feed, more valuable in nutrition than diet 2 that similar to natural detritus. The results show the intake phosphorus (IP) of G1 was significantly higher than that of G2, and both increased linearly with body size at a certain amount of diet. The retention phosphorus (RP) in fish of G1 was lower than G2. The relationship between retention phosphorus and body size was positive and stronger in G2. Significant difference in faecal phosphorus (FP) was found between G1 and G2. Body size significantly impacted the excretion phosphorus (EP) in G1 but G2. The loss of intake phosphor-us in G1 was 10.83-20.27 mg per g fish weight gain, higher than that in G2 for 6.63-9.56. Of the phosphor-us, about 10% was allocated into growth, 50% in faeces, and the rest lost in excretion. The main part of phosphorus was lost in faeces but excretion. The phosphorus budget of the fish could be described as 100IP = 7.40RP + 47.39FP + 36.63EP (Diet 1) or 100IP = 11.93RP + 56.64FP + 21.76EP (Diet 2).

Effects of diet, stocking density, and environmental factors on growth, survival, and metamorphosis of Manila clam Ruditapes philippinarum larvae

A series of experiments was conducted to evaluate the effects of diet, stocking density, and environmental factors on growth, survival, and metamorphosis of Manila clam Ruditapes philippinarum larvae. These experiments examined the following factors: diet (Isochrysts spp., Chlorella spp., and a mixture of Isochrysis spp. and Chlorella spp. [ 1: 1 w/w]), stocking density (5, 10, 15, and 20 larvae ml(-1)), light intensity (un-shaded, partially shaded, and fully shaded), water filtration (unfiltered and sand-filtered), water exchange (50% and 100% once every other day, 25%, 50%, and 100% once daily; 50% and 100% twice daily), and the use of substrate (with and without sand as the substrate). Results indicated that Chlorella spp. could replace 50% of Isochrysis spp. as a food source for the Manila clam larvae without affecting growth, survival, and metamorphosis. Larval growth decreased significantly with increasing stocking density. A density of 5-10 larvae ml(-1) appeared to be optimal for normal growth of Manila clam larvae. Neither diet nor stocking density used in the study had a significant effect on larval survival. Under partially shaded (light intensity = 1000-5000 lx) and fully shaded (light intensity <500 lx) conditions, larval growth was significantly faster than under direct sunlight (un-shaded). A water exchange rate of 50% twice daily provided optimum larval growth. Larvae grew significantly faster in the unfiltered water than in the sand-filtered water. Using sand as the substrate in the culture system significantly depressed the metamorphosis rate. The type and particle size of sand used as the substrate did not significantly affect growth and metamorphosis rates of the larvae. (C) 2005 Published by Elsevier B.V.

Diet shift of upland buzzards (Buteo hemilasius): evidence from stable carbon isotope ratios

We measured the stable carbon isotope ratios for muscle of the upland buzzards (Buteo hemilasius), plateau pika (Ochotoma curzoniae), root vole (Microtus oeconomus), plateau zokor (Myospalax fontanierii) and passerine bird species at the Haibei Alpine Meadow Ecosystem Research Station (HAMERS), and provided diet information of upland buzzards with the measurement of stable carbon isotopes in tissues of these consumers. The results showed that δ~(13)C values of small mammals and passerine bird species ranged from -25.57‰ to -25.78‰ (n = 12), and from -24.81‰ to -22.51% (n = 43), respectively, δ~(13)C values of the upland buzzards ranged from -22.60‰ to -23.10‰ when food was not available. The difference in δ~(13)C values (2.88‰±0.31‰) between upland buzzards and small mammals was much larger than the differences reported previously, 1‰-2‰, and showed significant difference, while 1.31‰±0.34‰ between upland buzzard and passerine bird species did not differ from the previously reported trophic fractionation difference of 1‰-2‰. Estimation of trophic position indicated that upland buzzards stand at trophic position 4.23, far from that of small mammals, i.e., upland buzzards scarcely captured small mammals as food at the duration of food shortage. According to isotope mass balance model, small mammals contributed 7.89% to 35.04% of carbon to the food source of the upland buzzards, while passerine bird species contributed 64.96% to 92.11%. Upland buzzards turned to passerine bird species as food during times of shortage of small mammals. δ~(13)C value, a useful indicator of diet, indicates that the upland buzzards feed mainly on passerine bird species rather than small mammals due to "you are what you eat" when small mammal preys are becoming scarce.

Summer diet of two sympatric species of raptors Upland Buzzard (Buteo hemilasius) and Eurasian Eagle Owl (Bubo bubo) in alpine meadow: problem of coexistence

Summer diets of two sympatric raptors Upland Buzzards (Buteo hemilasius Temminck et Schlegel) and Eurasian Eagle Owls (Bubo bubo L. subsp. Hemachalana Hume) were studied in an alpine meadow (3250 m a.s.l.) on Qinghai-Tibet Plateau, China. Root voles Microtus oeconomus Pallas, plateau pikas Ochotona curzoniae Hodgson, Gansu pikas O. cansus Lyon and plateau zokors Myospalax baileyi Thomas were the main diet components of Upland Buzzards as identified through the pellets analysis with the frequency of 57, 20, 19 and 4%, respectively. The four rodent species also were the main diet components of Eurasian Eagle Owls basing on the pellets and prey leftovers analysis with the frequency of 53, 26, 13 and 5%, respectively. The food niche breadth indexes of Upland Buzzards and Eurasian Eagle Owls were 1.60 and 1.77 respectively (higher value of the index means the food niche of the raptor is broader), and the diet overlap index of the two raptors was larger (C-ue = 0.90) (the index range from 0 - no overlap - to I - complete overlap). It means that the diets of Upland Buzzards and Eurasian Eagle Owls were similar (Two Related Samples Test, Z = -0.752, P = 0.452). The classical resource partitioning theory can not explain the coexistence of Upland Buzzards and Eurasian Eagle Owls in alpine meadows of Qinghai-Tibet Plateau. However, differences in body size, predation mode and activity rhythm between Upland Buzzards and Eurasian Eagle Owls may explain the coexistence of these two sympatric raptors.

Optimal Fuzzy PID Controller with Incomplete Derivation and Its Simulation Research on Application of Intelligent Artificial Legs

提出了一种新的最优模糊PID控制器，它由两部分组成，即在线模糊推理机构和带有不完全微分的常规PID控制器，在模糊推理机构中，引入了三个可调节因子xp,xi和xd，其作用是进一步修改和优化模糊推理的结果，以使控制器对一个给定对象具有最优的控制效果，可调节因子的最优值采用ITAE准则及Nelder和Mead提出的柔性多面体最优搜索算法加以确定，这种PID控制器被用来控制由作者设计的智能人工腿中的一个直流电机，仿真结果表明该控制器的设计是非常有效的，它可被用于控制各种不同的对象和过程。