Growth kinetics of 1-2 mm and 3-4 mm colonies of Nostoc sphaeroides (Cyanophyta) in outdoor culture

Nostoc sphaeroides Kutzing was cultivated in paddlewheel-driven raceway ponds and the growth kinetics of 1-2 mm and 3-4 mm colonies of N. sphaeroides was studied. The biomass productivities in 2.5 m(2) raceway ponds inoculated with 1-2 mm and 3-4 mm colonies were 5.2 and 0.25 g dry wt m(-2) stop d(-1), respectively. Furthermore, differently sized colonies showed different relative water content, total soluble carbohydrates, chlorophyll a content and density of filaments. This is the first report on mass culture of N. sphaeroides under outdoor conditions.

The accumulation and metabolism of astaxanthin in Scenedesmus obliquus (chlorophyceae)

The biosynthesis and metabolism of astaxanthin in coenobium alga Scenedesmus obliquus were investigated using a two-stage culture. The first stage was for the analysis of biosynthesis and accumulation of astaxanthin in alga cells which were cultured under induction conditions (incubation at 30 degrees C and illumination of 180 mu mol m(-2) s(-1)) for 48 h. The composition of the secondary carotenoids in algal cells was analyzed and seven ketocarotenoids were identified. The results implied that S. obliquus synthesized astaxanthin from beta-carotene through three possible pathways. In the second stage, the cultures were transferred to normal conditions (incubation at 25 C and illumination of 80 mu mol m(-2) s(-1)) for 72 h. Algal cells accumulated more chlorophyll and biosynthesis of secondary carotenoids terminated, the content of secondary carotenoids decreased from 59.48 to 6.57%. The results inferred that accumulation and metabolism of astaxanthin could be controlled by cultivated conditions which also could lead the mobilization of secondary carotenoids to support the algal cell growth. The results also implied that presumed conversions from astaxanthin to lutein or antheraxanthin could be modulated by culturing conditions. (C) 2008 Published by Elsevier Ltd.

Avian influenza virus infection induces differential expression of genes in chicken kidney

The pathogenic process of highly pathogenic avian influenza virus (HPAIV) infection is poorly understood. To explore the differential expression of kidney genes as a result of HPAIV infection, two cDNA libraries were constructed from uninfected and infected kidneys by suppression subtractive hybridization (SSH). Fifteen genes including IFN-stimulated genes (ISG12), lymphocyte antigen 6 complex locus E gene (LY6E), matrix Gla protein gene (MGP), lysozyme gene, haemopoiesis related membrane protein I gene, KIAA1259, MGC68696, G6pe-prov protein gene (G6PC), MGC4504, alcohol dehydrogenase gene (ADH), glutathione S-transferase gene (GST), sodium-dependent high-affinity dicarboxylate transporter gene (SDCT), Synaptotagmin XV (SytXV) and two novel genes were found significantly up-regulated or dramatically suppressed. Differential expression of these genes was further identified by Northern blot. Functional analysis indicated that the regulation of their expression might contribute to the pathogenic process of HPAIV infection. In contrast, the increased expression of three IFN-stimulated genes named ISG12, LY6E, and haemopoiesis related membrane protein 1 gene might reflect host defense responses. Further study showed that ISG12 protein failed to directly interact with NS1 protein of HPAIV which expressed simultaneously in the organs where HPAIV replication occurred, by use of BacterioMatch two-hybrid system. Therefore, our findings may provide new insights into understanding the molecular mechanism underlying the pathophysiological process of HPAIV infection in chicken. (c) 2007 Elsevier Ltd. All rights reserved.

Dietary phosphorus requirement of juvenile black seabream, Sparus macrocephalus

A growth trial was conducted to estimate the optimum requirement of dietary available phosphorus (P) for black seabream (Sparus macrocephalus) in indoor net cages (1.5x1.0x1.0 m). Triplicate groups of black seabream (11.45 +/- 0.02 g) were fed diets containing graded levels (0.18, 0.36, 0.54, 0.72, 0.89 and 1.07%) of available P to satiation for 8 weeks. The basal diet (diet 1), containing 0.18% available P, was supplemented with graded levels of monosodium phosphate (NaH2PO4 2H(2)O) to formulate five experimental diets. The fish were fed twice daily (08:00 h and 16:00 h) and reared in seawater (salinity, 26-29 g l(-1)) at a temperature of 28 +/- 1 degrees C. Dissolved oxygen during the experiment was above 5 mg l(-1). The specific growth rate (SGR), weight gain (WG), feed efficiency (FE) and protein efficiency ratio (PER) were all significantly improved by dietary phosphorus up to 0.54% (P<0.05) and then leveled off beyond this level. Hepatosomatic index (HSI) was inversely correlated with dietary phosphorus levels (P< 0.05). Efficiency of P utilization stabled in fish fed diets containing 0.18%-0.54% available P and then decreased dramatically with further supplementation of dietary phosphorus. Body composition analysis showed that the whole-body lipid, ash, calcium and phosphorus contents were all significantly affected by dietary available P concentration (P<0.05), however, no significance were found in whole-body calcium/phosphorus (Ca/P) ratios among all the treatments (P>0.05). Dietary phosphorus levels also affected the mineralization of vertebrae, skin and scale (P<0.05). Ca/P ratios in vertebrae and scale were not influenced by dietary P supplementation, while skin Ca/P ratio increased statistically with dietary available P levels (quadratic effect, P<0.001). The blood chemistry analysis showed that dietary available P had distinct effects on enzyme activities of alkaline phosphatase (ALP) and plasma lysozyme (LSZ), as well as contents of triacyglycerol (TG) and total cholesterol (T-CHO) (P<0.05). Broken-line analysis showed maximum weight gain (WG) was obtained at dietary available P concentrations of 0.55%. Quadratic analysis based on P contents in whole fish, vertebrae or scale indicated that the requirements were 0.81, 0.87 and 0.88%, respectively. Signs of phosphorus deficiency were characterized by poor growth, slightly reduced mineralization and an increase in body lipid content. (C) 2008 Published by Elsevier B.V.

Structure, organization and expression of common carp (Cyprinus carpio L.) SLP-76 gene

SLP-76 is an important member of the SLP-76 family of adapters, and it plays a key role in TCR signaling and T cell function. Partial cDNA sequence of SLP-76 of common carp (Cyprinus carpio L.) was isolated from thymus cDNA Library by the method of suppression subtractive hybridization (SSH). Subsequently, the full length cDNA of carp SLP-76 was obtained by means of 3' RACE and 5' RACE, respectively. The full Length cDNA of carp SLP-76 was 2007 bp, consisting of a T-terminal untranslated region (UTR) of 285 bp, a T-terminal. UTR of 240 bp, and an open reading frame of 1482 bp. Sequence comparison showed that the deduced amino acid sequence of carp SLP-76 had an overall similarity of 34-73% to that of other species homotogues, and it was composed of an NH2-terminal domain, a central proline-rich domain, and a C-terminal SH2 domain. Amino acid sequence analysis indicated the existence of a Gads binding site R-X-X-K, a 10-aa-long sequence which binds to the SH3 domain of LCK in vitro, and three conserved tyrosine-containing sequence in the NH2-terminal domain. Then we used PCR to obtain a genomic DNA which covers the entire coding region of carp SLP-76. In the 9.2 k-long genomic sequence, twenty one exons and twenty introns were identified. RT-PCR results showed that carp SLP-76 was expressed predominantly in hematopoietic tissues, and was upregulated in thymus tissue of four-month carp compared to one-year old carp. RT-PCR and virtual northern hybridization results showed that carp SLP-76 was also upregulated in thymus tissue of GH transgenic carp at the age of four-months. These results suggest that the expression level of SLP-76 gene may be related to thymocyte development in teleosts. (c) 2007 Published by Elsevier Ltd.

Nitrite utilization by Chaetoceros muelleri under elevated CO2 concentration

Chaetoceros muelleri (Lemn.) was cultured with nitrite (NO2-) or nitrate (NO3-) as the sole nitrogen source and aerated with air or with CO2-enriched air. Cells of C. muelleri excreted into the medium nitrite produced by reduction of nitrate when grown with 100 mu M NaNO3 as nitrogen source. Accordingly, NO2- concentration reached 10.4 mu M after 95 h at the low CO2 condition (aerated with air); while the maximum NO2- concentration was only around 2.0 mu M at the high CO2 condition (aerated with 5% CO2 in air), furthermore, after 30 h it decreased to no more than 1.0 mu M. NO2- was almost assimilated in 80 h when C. muelleri was cultured at the high CO2 condition with 100 mu M NaNO2 as sole nitrogen source. At the high CO2 condition, after 3 h the activity of nitrite reductase was as much as 50% higher than that at the low CO2 condition. It was indicated that enriched CO2 concentration could inhibit nitrite excretion and enhance nitrite assimilation by cells. Therefore, aeration with enriched CO2 might be an effective way to control nitrite content in aquaculture systems.

Toll-like receptor 3 regulates Mx expression in rare minnow Gobiocypris rarus after viral infection

Toll-like receptor 3 (TLR3) plays a key role in activating immune responses during viral infection. To study the genes involved in the regulatory function of TLR3 in the rare minnow Gobiocypris rarus after viral infection, a full-length cDNA of TLR3 (GrTLR3) with a splice variant (GrTLR3s) was identified by homologous cloning and RACE techniques. The antiviral effector molecule Mx gene was cloned and partially sequenced. The mRNA expression levels of GrTLR3, GrTLR3s, and Mx were studied in different tissues before and after virus infection by real-time quantitative RT-PCR. The transcripts of all three genes in liver were significantly increased following GCRV infection (P<0.05). The mRNA levels in liver were upregulated at 24 h post-injection for GrTLR3 and GrTLR3s, and at 12 h for Mx. The upregulated expression levels were several folds for GrTLR3s, tens of folds for GrTLR3, and hundreds of folds for Mx. By semi-quantitative RT-PCR, GrTLR3 and Mx expressed at all the developmental stages, whereas GrTLR3s could only be detected at later developmental stages. Using RNAi and transgenic techniques, GrTLR3 mediated Mx expression but GrTLR3s did not. The time-dependent upregulation of receptor and effector, and the Mx over-expression dependent on TLR3, indicated that GrTLR3 regulated Mx expression in viral infection through a configuration change in rare minnow, and its splice variant did not contribute to the process.

Molecular cloning and characterization of common carp (Cyprinus carpio L.) TCR gamma and CD3 gamma/delta chains

Partial cDNA sequences of TCR gamma and CD3 gamma/delta were isolated from the thymus of common carp (Cyprinus carpio L.) by the method of suppression subtractive hybridization (SSH). Subsequently the full length cDNAs of carp TCR gamma and CD3 gamma/delta were obtained by means of 3' RACE and 5' RACE, respectively. The full length of carp TCR gamma chain is 1368 bp and encodes 326 amino acids including a signal peptide region of 19 amino acids and a transmembrane region of 23 amino acids at the C-terminal region from aa 291 to 313. The V region of carp TCR gamma contains 109 amino acids, the core motif FGXG in J segment was also found in carp TCR gamma. The C region of carp TCR gamma contains the characteristic CX6PX6WX45C motif. The CP region of carp TCR C gamma contains 37 amino acids. The full length of carp CD3 gamma/delta is 790 bp and encodes 175 amino acids including a signal peptide region of 17 amino acids and a transmembrane region of 23 amino acids from aa 93 to 115. Similar to other known CD3 gamma/delta s, four cysteine residues in the extracellular domain and an immunoreceptor tyrosine-based activation motif ITAM (YxxL/Ix6-8YxxL/I) in the intracellular domain are also included in carp CD3 gamma/delta. Differing from other known CD3 gamma/delta s, carp CD3 gamma/delta tacks the CXXCXE motif in the extracellular domain. RTPCR analysis demonstrated that the expression of TCR gamma gene was mainly in the thymus and gill of 6-month carp, but in 18-month carp, TCR gamma gene was detected in all the examined tissues. The expression of CD3 gamma/delta gene was detected in all examined tissues of 6 and 18-month carp; among them, the highest expression level was in the thymus of 6-month carp. In situ hybridization showed that CD3 gamma/delta-expressing cells were widely distributed in the head kidney, spleen and kidney of carp, whereas in the thymus, they were densely distributed in the lymphoid outer zone and scattered in the epithelioid inner zone. (c) 2007 Published by Etsevier Ltd.

Identification and characterization of a novel gene differentially expressed in zebrafish cross-subfamily cloned embryos

m Background: Cross-species nuclear transfer has been shown to be a potent approach to retain the genetic viability of a certain species near extinction. However, most embryos produced by cross-species nuclear transfer were compromised because that they were unable to develop to later stages. Gene expression analysis of cross-species cloned embryos will yield new insights into the regulatory mechanisms involved in cross-species nuclear transfer and embryonic development. Results: A novel gene, K31, was identified as an up-regulated gene in fish cross-subfamily cloned embryos using SSH approach and RACE method. K31 complete cDNA sequence is 1106 base pairs (bp) in length, with a 342 bp open reading frame (ORF) encoding a putative protein of 113 amino acids (aa). Comparative analysis revealed no homologous known gene in zebrafish and other species database. K31 protein contains a putative transmembrane helix and five putative phosphorylation sites but without a signal peptide. Expression pattern analysis by real time RT-PCR and whole-mount in situ hybridization (WISH) shows that it has the characteristics of constitutively expressed gene. Sub-cellular localization assay shows that K31 protein can not penetrate the nuclei. Interestingly, over-expression of K31 gene can cause lethality in the epithelioma papulosum cyprinid (EPC) cells in cell culture, which gave hint to the inefficient reprogramming events occurred in cloned embryos. Conclusion: Taken together, our findings indicated that K31 gene is a novel gene differentially expressed in fish cross-subfamily cloned embryos and over-expression of K31 gene can cause lethality of cultured fish cells. To our knowledge, this is the first report on the determination of novel genes involved in nucleo-cytoplasmic interaction of fish cross-subfamily cloned embryos.

Responses of photosynthetic activity in the drought-tolerant cyanobacterium, Nostoc flagelliforme to rehydration at different temperature

Photosynthetic activity during rehydration at four temperatures (5, 15, 25, 35 degrees C) was studied in a terrestrial, highly drought-tolerant cyanobacterium, Nostoc flagelliforme. At all the temperatures, the optimum quantum yield F-v/F-m increased rapidly within I It and then increased slowly during the process of rehydration. The increase in F-v/F-m at 25 and 35 degrees C was larger than that at 5 and 15 degrees C. In addition, the changes of initial intensity of fluorescence (F-0) and variable fluorescence (F-v) were more significant at 25 and 35 degrees C than those at 5 and 15 degrees C. Chlorophyll a content increased with the increase of temperature during the course of rehydration, with this being more pronounced at 25 and 35 degrees C. The photosynthetic rates at 25 and 35 degrees C were higher than those at 5 and 15 degrees C. Induction of chlorophyll fluorescence with sustained rewetting at 5 and 15 degrees C had two phases of transformation, whereas at 25 and 35 degrees C it had a third peak kinetic phase and showed typical chlorophyll fluorescence steps on rewetting for 24 h, representing a normal physiological state. A comparison of the chlorophyll fluorescence parameters, chlorophyll a content, and the chlorophyll fluorescence induction led to the conclusion that N. flagelliforme had a more rapid and complete recovery at 25 and 35 degrees C than that at 5 and 15 degrees C, although it could recover its photosynthetic activity at any of the four temperatures. (c) 2007 Published by Elsevier Ltd.

Control of lunar and martian dust - Experimental insights from artificial and natural cyanobacterial and algal crusts in the desert of Inner Mongolia, China

Studies on the colonization of environmentally extreme ground surfaces were conducted in a Mars-like desert area of Inner Mongolia, People's Republic of China, with microalgae and cyanobacteria. We collected and mass-cultured cyanobacterial strains from these regions and investigated their ability to form desert crusts artificially. These crusts had the capacity to resist sand wind erosion after just 15 days of growth. Similar to the surface of some Chinese deserts, the surface of Mars is characterized by a layer of fine dust, which will challenge future human exploration activities, particularly in confined spaces that will include greenhouses and habitats. We discuss the use of such crusts for the local control of desert sands in enclosed spaces on Mars. These experiments suggest innovative new directions in the applied use of microbe-mineral interactions to advance the human exploration and settlement of space.

A simple closed aquatic ecosystem (CAES) for space

A closed aquatic ecosystem (CAES) was developed to stud), the effects of microgravity on the function of closed ecosystems aboard the Chinese retrieved satellite and on the spacecraft SHENZHOU-II. These systems housed a small freshwater snail (Bulinus australianus) and an autotrophic green algae (Chlorella pyrenoidosa). The results of the test on the satellite were that the concentration of algae changed little, but that the snails died during the experiments. We then sought to optimize the function of the control system, the cultural conditions and the data acquisition system and carried out an experiment on the spacecraft SHENZHOU-II. Using various sensors to monitor the CAES, real-time data regarding the operation of the CAES in microgravity was acquired. In addition, all on-board Ig centrifuge was included to identify gravity-related factors. It was found that microgravity is the major factor affecting the operation of the CAES in space. The change in biomass of the primary producer during each day in microgravity was larger than that of the control groups. The mean biomass concentration per day in the microgravity group decreased, but that of the control groups increased for several days and then leveled off. Space effects on the biomass of a primary producer may be a result of microgravity effects leading to increasing metabolic rates of the consumer combined with decreases in photosynthesis. (c) 2007 COSPAR. Published by Elsevier Ltd. All rights reserved.

A novel chemiluminescent immunoassay for microcystin (MC) detection based on gold nanoparticles label and its application to MC analysis in aquatic environmental samples

A novel chemiluminescent immunoassay method based on gold nanoparticles was developed for the detection of microcystins (MCs). The immunoassay included three main steps: indirect competitive immunoreaction, oxidative dissolution of gold nanoparticles, and indirect determination for MCs with Au3+-catalysed luminol chemiluminesent system. The method has a wide working range (0.05-10 mu g L-1, r(2) = 0.9914), the limit of detection was determined to be 0.024 mu g L-1, which is much lower than the World Health Organization's proposed guidelines (1 mu g L-1) for drinking-water. The proposed method was applied to MC analysis in natural water and fish tissue samples, and most results in the proposed method were in agreement with the conventional indirect competitive enzyme-linked immunosorbent assay method, which indicated that the new chemiluminescent immunoassay was sensitive, reliable, and suitable for MC analysis in natural water and fish tissue samples.

The response of antioxidant systems in Nostoc sphaeroides against UV-B radiation and the protective effects of exogenous antioxidants

UV radiation is one of many harmful factors found in space that are detrimental to organisms on earth in space exploration. In the present work, we examined the role of antioxidant system in Nostoc sphaeroides Kutz (Cyanobacterium) and the effects of exogenously applied antioxidant molecules on its photosynthetic rate under UV-B radiation. It was found that UV-B radiation promoted the activity of antioxidant system to protect photosystem 11 (PSII) and exogenously applied antioxidant: sodium nitroprusside (SNP) and N-acetylcysteine (NAC) had an obvious protection on PSII activity under UV-B radiation. The activity of superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), peroxidase (POD, EC 1.11.1.7) and content of NIDA (malondialdehyde) and ASC (ascorbate) were improved by 0.5 mM and 1 mM SNP, but 0.1 mM SNP decreased the activity of antioxidant system. Addition of exogenous NAC decreased the activity of SOD, POD, CAT and the content MDA and ASC. In contrast, exogenously applied NAC increased GSH content. The results suggest that exogenous SNP and NAC may protect algae by different mechanisms: SNP may play double roles as both sources of reactive free radicals as well as ROS scavengers in mediating the protective role of PSII on algae under UV-B radiation. On the other hand, NAC functions as an antioxidant or precursor of glutathione, which could protect PSII directly from UV-B radiation. (c) 2007 COSPAR, Published by Elsevier Ltd. All rights reserved.

Description of a new genus and two new species of labeonine fishes from South China (Teleostei : Cyprinidae)

A new genus and two new species are described from the Pearl River drainage in Guangxi Province, South China. Hongshuia, new genus, can be distinguished from all other Asian genera of the Labeonini by having a lower lip with its median lobe modified into a round, fleshy plate peripherally greatly protruded so as to form a ring-like fold that is posteromedially continuous with the mental region, and centrally sunken so as to form a round, flat, fleshy pad. This genus is distinct from all other Asian labeonine genera of the Garrina except for one newly described species of Parasinilabeo ( P. longibarbus), Pseudocrossocheilus, and Sinocrossocheilus, in the presence of well-developed maxillary barbels. Hongshuia differs from the above three genera in the lower lip morphology, and further from both Pseudocrossocheilus and Qianlabeo in the number of pharyngeal tooth rows and from Sinocrossocheilus in the colour pattern. Two new species, H. banmo and H. paoli, differ in the distribution density and degree of development of papillae on the rostral fold, depth of indentations on the distal edge of the rostral fold, presence or absence of papillae on the lower lip, size and shape of tubercles on the tip of the snout and anterior portion of the lachrymal, length, position and colour pattern of the dorsal fin, and snout length.