Characteristics of the microbial communities in the integrated vertical-flow constructed wetlands

Microorganisms play an important role in removing pollutants from constructed wetlands. We investigated the microbial characteristics in a novel integrated vertical-flow constructed wetland (IVCW), which has been in operation in Wuhan, China since 1998. We used phospholipid fatty acid (PLFA) and amoA gene to analyze the structure and diversity of the microbial community within the IVCW. PLFA results suggested that the amount of bacterial PLFA was significantly higher than that of fungal PLFA, but the total microbial biomass represented by PLFA index was low in the system. Microbial spatial distribution showed significantly higher bacterial (both G(+) and G(-)) and fungal biomass in the surface than in the subsurface layers. The ratios of monounsaturated to branched PLFA demonstrated that an anaerobic layer sandwiched by two aerobic layers existed in the IVCW, consistent with the redox potential results. Analysis of the amoA revealed the presence of Nitrosomonas-like sequences in the surface substrate of the downflow chamber and apparent diversities of ammonia-oxidizing bacteria in the system. These results suggest that microorganisms, despite their relatively low biomass, have inhabited the IVCW, and the results will offer some valuable information on microbe to system designers and managers.

Phosphorus in interstitial water induced by redox potential in sediment of Dianchi Lake, China

The sediment redox potential was raised in the laboratory to estimate reduction of internal available phosphorus loads, such as soluble reactive phosphorus (SRP) and total phosphorus (TP), as well as the main elements of sediment extracts in Dianchi Lake. Several strongly reducing substances in sediments, which mainly originated from anaerobic decomposition of primary producer residues, were responsible for the lower redox potential. In a range of -400 to 200 mV raising the redox potential of sediments decreased TP and SRP in interstitial water. Redox potentials exceeding 320 mV caused increases in TP, whereas SRP maintained a relatively constant minimum level. The concentrations of Al, Fe, Ca2+, Mg2+, K+, Na+ and S in interstitial water were also related to the redox potential of sediments, suggesting that the mechanism for redox potential to regulate the concentration of phosphorus in interstitial water was complex.

Enhancing the carbonation of MgO cement porous blocks through improved curing conditions

The use of reactive magnesia (MgO) as the binder in porous blocks demonstrated significant advantages due to its low production temperatures and ability to carbonate, leading to significant strengths. This paper investigates the enhancement of the carbonation process through different curing conditions: water to cement ratio (0.6-0.9), CO2 concentration (5-20%), curing duration (1-7 days), relative humidity (55-98%), and wet/dry cycling frequency (every 0-3 days), improving the carbonation potential through increased amounts of CO2 absorbed and enhanced mechanical performance. UCS results were supported with SEM, XRD, and HCl acid digestion analyses. The results show that CO2 concentrations as low as 5% can produce the required strengths after only 1 day. Drier mixes perform better in shorter curing durations, whereas larger w/c ratios are needed for continuous carbonation. Mixes subjected to 78% RH outperformed all the others, also highlighting the benefits of incorporating wet/dry cycling to induce carbonation. © 2014 Elsevier Ltd.

Contributions of phosphatase and microbial activity to internal phosphorus loading and their relation to lake eutrophication

Phosphatase may accelerate the process of lake eutrophication through improving phosphorus bioavailability. This mechanism was studied in three Chinese eutrophic shallow lakes (Lake Taihu, Lake Longyang,and Lake Lianhua). Phosphatase activity was related to the concentration of soluble reactive phosphorus (SRP) and chlorophyll a. Stability of dissolved phosphatase in reverse micelles may be attributed to molecular size, conformation and active residues of the enzyme. At the site with Microcystis bloomed in Lake Taihu, dissolved phosphatase activity was higher and more stable in micelles, SRP concentrations were lower in interstitial water, the contents of different forms of phosphorus and the amounts of aerobic bacteria were lower while respiration efficiency was higher in sediments. Phosphobacteria, both inorganic and organic and other microorganisms were abundant in surface water but rare in sediments. Therefore, internal phosphorus may substantially flux into water column by enzymatic hydrolysis and anaerobic release, together with mobility of bacteria, thereby initiating the bloom. In short, biological mechanism may act in concert with physical and chemical factors to drive the internal phosphorus release and accelerate lake eutrophication.

Genetic structure and low-genetic diversity suggesting the necessity for conservation of the Chinese Longsnout catfish, Leiocassis longirostris (Pisces : Bagriidae)

Genetic variation and phylogenetic relationship of Leiocassis longirostris populations from the Yangtze River were investigated at mitochondrial DNA level. The samples were collected from the upstream and mid-downstream of the Yangtze River. Three mitochondrial DNA fragments, ND5/6, cytochrome b (Cyt b) and control region (D-loop), were amplified and then digested by 10 restriction endonucleases. Twenty-three D-loop fragments randomly selected were sequenced. Digestion patterns of ND5/6 by AluI and HaeIII, D-loop by HinfI and RsaI, and Cyt b by HaeIII were polymorphic. Ten and eighteen haplotypes were obtained from RFLP data and sequence data, respectively. The individuals from upstream and mid-downstream of the Yangtze River were apparently divided into two groups. The average genetic distance was 0.008 and 0.010 according to the two data. Low diversities and decreasing abundance indicated that Leiocassis longirostris may be in severe danger and reasonable measures of fishery management should be taken.

The functional divergence of two glgP homologues in Synechocystis sp PCC 6803

Glycogen phosphorylase (GlgP, EC 2.4.1.1) catalyzes the cleavage of glycogen into glucose-1-phosphate (Glc-1-P), the first step in glycogen catabolism. Two glgP homologues are found in the genome of Synechocystis sp. PCC 6803, a unicellular cyanobacterium: sll1356 and slr1367. We report on the different functions of these glgP homologues. sll1356, rather than slr1367, is essential for growth at high temperatures. On the other hand, when CO2-fixation and the supply of glucose are both limited, slr1367 is the key factor in glycogen metabolism. In cells growing autotrophically, sll1356 plays a more important role in glycogen digestion than slr1367. This functional divergence is also supported by a phylogenetic analysis of glgP homologues in cyanobacteria.

Efficient RNA interference in zebrafish embryos using siRNA synthesized with SP6 RNA polymerase

Double-stranded RNA (dsRNA) has been shown to be a useful tool for silencing genes in zebrafish (Danio rerio), while the blocking specificity of dsRNA is still of major concern for application. It was reported that siRNA (small interfering RNA) prepared by endoribonuclease digestion (esiRNA) could efficiently silence endogenous gene expression in mammalian embryos. To test whether esiRNA could work in zebrafish, we utilized Escherichia coli RNaseIII to digest dsRNA of zebrafish no tail (ntl), a mesoderm determinant in zebrafish and found that esi-ntl could lead to developmental defects, however, the effective dose was so close to the toxic dose that esi-ntl often led to non-specific developmental defects. Consequently, we utilized SP6 RNA polymerase to produce si-ntl, siRNA designed against ntl, by in vitro transcription. By injecting in vitro synthesized si-ntl into zebrafish zygotes, we obtained specific phenocopies of reported mutants of ntl. We achieved up to a 59%no tail phenotype when the injection concentration was as high as 4 mu g/mu L. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and whole-mount in situ hybridization analysis showed that si-ntl could largely and specifically reduce mRNA levels of the ntl gene. As a result, our data indicate that esiRNA is unable to cause specific developmental defects in zebrafish, while siRNA should be an alternative for downregulation of specific gene expression in zebrafish in cases where RNAi techniques are applied to zebrafish reverse genetics.

Cloning of biologically active genomes from a Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus isolate by using a bacterial artificial chromosome

Purification of genotypes from baculovirus isolates provides understanding of the diversity of baculoviruses and may lead to the development of better pesticides. Here, we report the cloning of different genotypes from an isolate of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HaSNPV) by using a bacterial artificial chromosome (BAC). A transfer vector (pHZB10) was constructed which contained an Escherichia coli mini-F replicon cassette within the upstream and downstream arms of HaSNPV polyhedrin gene. Hz2e5 cells were co-transfected with wild-type HaSNPV DNA and pHZB10 to generate recombinant viruses by homologous recombination. The DNA of budded viruses (BVs) was used to transform E. coli. One of the bacmid colonies, HaBacHZ8, has restriction enzyme digestion profiles similar to an in vivo cloned strain HaSNPV-G4, the genome of which has been completely sequenced. For testing the oral infectivity, the polyhedrin gene of HaSNPV was reintroduced into HaBacHZ8 to generate the recombinant bacmid HaBacDF6. The results of one-step growth curves, electron microscopic examination, protein expression analysis and bioassays indicated that HaBacDF6 replicated as well as HaSNPV-G4 in vitro and in vivo. The biologically functional HaSNPV bacmids obtained in this research will facilitate future studies on the function genomics and genetic modification of HaSNPV. (C) 2003 Elsevier B.V. All rights reserved.

Genetic divergence between Cyprinus carpio carpio and Cyprinus carpio haematopterus as assessed by mitochondrial DNA analysis, with emphasis on origin of European domestic carp

Although common carp is the major fish species in Asian and European aquaculture and many domestic varieties have occurred, there is a controversy about the origination of European domestic common carp. Some scientists affirmed that the ancestor of European domestic common carp was Danube River wild common carp, but others considered it might be Asian common carp. For elucidating origination of European domestic common carp, we chose two representative European domestic common carp strains (German mirror carp and Russian scattered scaled mirror carp) and one wild common carp strain of Cyprinus carpio carpio subspecies (Volga River wild common carp) and two Asian common carp strains, the Yangtze River wild common carp (Cyprinus carpio haematopterus) and traditionally domestic Xingguo red common carp, as experimental materials. ND5-ND6 and D-loop segments of mitochondrial DNA were amplified by polymerase chain reaction and analyzed through restriction fragment length polymorphism (RFLP) and sequencing respectively. The results revealed that HaeIII and DdeI digestion patterns of ND5-ND6 segment and sequences of control region were different between European subspecies C. carpio carpio and Asian subspecies C. carpio haematopterus. Phylogenetic analysis showed that German mirror carp and Russian scattered scaled mirror carp belonged to two subspecies, C. carpio carpio and C. carpio haematopterus, respectively. Therefore, there were different ancestors for domestic carp in Europe: German mirror carp was domesticated from European subspecies C. carpio carpio and Russian scattered scaled mirror carp originated from Asian subspecies C. carpio haematopterus.

In vitro culture of embryonic hearts from guppy fish (Poecilia reticulata)

Forty embryonic hearts were taken out by anatomical needle from denuded embryos of the ovoviviparity guppy fish that were dechorioned by mechanic method or by trypsin digestion, and were in vitro cultured. In the cultured hearts, 80% have maintained beating in vitro for 4 weeks, and the longest record for beating was 142 d. Owing to fish embryo transparency, beating frequency and blood color changes are easily viewed from the embryonic hearts under a dissecting microscope. The current study established the in vitro culture method of embryonic hearts in guppy fish, which can be used as a model for the study of heart and cardiovascular system in vertebrates.

Quantitative structure-property relationship study on reductive dehalogenation of selected halogenated aliphatic hydrocarbons in sediment slurries

In this study, by the use of partial least squares (PLS) method and 26 quantum chemical descriptors computed by PM3 Hamiltonian, a quantitative structure-property relationship (QSPR) model was developed for reductive dehalogenation rate constants of 13 halogenated aliphatic compounds in sediment slurry under anaerobic conditions. The model can be used to explain the dehalogenation mechanism. Halogenated aliphatic compounds with great energy of the lowest unoccupied molecular orbital (E-lumo), total energy (TE), electronic energy (EE), the smallest bond order of the carbon-halogen bonds (BO) and the most positive net atomic charges on an atom of the molecule (q(+)) values tend to be reductively dehalogenated slow, whereas halogenated aliphatic compounds with high values of molecular weight (Mw), average molecular polarizability (a) and core-core repulsion energy (CCR) values tend to be reductively dehalogenated fastest. (C) 2001 Published by Elsevier Science Ltd.

Transgenes in F-4 pMThGH-transgenic common carp (Cyprinus carpio L.) are highly polymorphic

To gain information on the integration pattern of pMThGH-transgene, 50 transgenes were recovered from F-4 generation of pMThGH transgenic common carp (Cyprinus carpio L,) and 33 recovered genes were analyzed. The restriction maps of these recovered genes were constructed by digestion with five kinds of enzymes. These transgenes can be classified into 4 types according to their restriction maps. Only one type of transgenes maintains its original molecular form, whereas the other three types are very different from the original one and vary each other on both molecular weight and restriction maps. This implies that the sequences of most transgenes have been deleted and/or rearranged during integration and inheritance. The results of PCR amplification and Southern blot hybridization indicate that MThGH in Type I transgene keeps intact but most of its sequence has been deleted in other three types. All these results suggest that transgenes in F-4 generation of transgenic carp are highly polymorphic. Two DNA fragments concerning integration site of transgenes were cloned from recovered transgenes, and found to be homologous to the 5'UTR of beta -actin gene of common carp and mouse mRNA for receptor tyrosine kinase (RTK), respectively.

Study of sorption, biodegradation and isomerization of HCH in stimulated sediment/water system

This paper reported the sorption, biodegradation and isomerization of hexachlorocyclohexane (HCH) in laboratory sediment/water system under aerobic and anaerobic conditions, respectively. The effect of organic nutrient addition to the sorption of HCH was also investigated. It indicates that HCH is highly adsorbed on sediments under both conditions. During the tests, the biodegradation and isomerization of HCH were dramatically speeded up after organic nutrient additions, especially in the case of the observation under aerobic condition. It was found, beta-HCH was the most persistent in the environment, that is due to the isomerization of alpha-HCH in a big amount to beta-HCH, besides its chemical stability. (C) 1997 Elsevier Science Ltd.

城市生活有机垃圾厌氧降解的过程机理与工程应用研究

首先进行城市生活有机垃圾典型组分的厌氧发酵产甲烷和产氢特性研究，在此基础上，设计厌氧发酵联产氢气和甲烷的组合工艺提高能源回收效率，并采用厨余垃圾和废纸联合厌氧消化的方式避免厨余垃圾单独厌氧消化的挥发性脂肪酸抑制。其次，结合国内近年出现的城市生活垃圾分选技术，分别以机械干分选有机垃圾和水分选有机垃圾为原料进行厌氧发酵产甲烷实验，并根据实验结果设计日处理500吨城市生活垃圾厌氧沼气工程及进行经济性评价。主要结论如下：（1）糖和淀粉类的生化产甲烷能力为260 mL/gVS，纤维素，粗纤维、蛋白类和脂类分别为244、145、258 、757 mL/gVS；蛋白质类原料在厌氧消化过程中容易形成“抑制型稳态”；脂类原料容易导致长链脂肪酸抑制。（2）碳水化合物（糖、淀粉和纤维素）是最佳的厌氧发酵制氢原料，蛋白类、脂类和木质纤维类均不适宜作为厌氧发酵制氢原料。采用厌氧发酵联产氢气和甲烷的组合工艺可以显著提高能源回收率。（3）厨余垃圾单独厌氧发酵容易受到VFAs的强烈抑制，采用厨余垃圾与废纸联合厌氧发酵，能够避免VFAs抑制。（4）水分选有机垃圾的生物可降解性优于机械干分选有机垃圾，在原料TS浓度为11%~16%时的甲烷产率为273~314 L/kgVS。（5）国家财政补贴，税收优惠和CDM额外收益决定了城市生活垃圾的厌氧消化与热电肥联产工程的经济可行性。

Fatty-acids and their C-13 signatures in seep carbonates from hydrocarbon seeps on the upper (GC 185) and lower (AC 645) continental slope of the Gulf of Mexico

Here we reported the fatty-acids and their δ 13C values in seep carbonates collected from Green Canyon lease block 185 (GC 185; Sample GC-F) at upper continental slope (water depth: ∼540 m), and Alaminos Canyon lease block 645 (GC 645; Sample AC-E) at lower continental slope (water depth: ∼2200 m) of the Gulf of Mexico. More than thirty kinds of fatty acids were detected in both samples. These fatty acids are maximized at C16. There is a clear even-over-odd carbon number predominance in carbon number range. The fatty acids are mainly composed of n-fatty acids, iso-/anteiso-fatty acids and terminally branched odd-numbered fatty acids (iso/anteiso). The low δ 13C values (−39.99‰ to.32.36‰) of n-C12:0, n-C13:0, i-C14:0and n-C14:0 suggest that they may relate to the chemosynthetic communities at seep sites. The unsaturated fatty acids n-C18:2 and C18:1Δ9 have the same δ 13C values, they may originate from theBeggiatoa/Thioploca. Unlike other fatty acids, the terminally branched fatty acids (iso/anteiso) show lowerδ 13C values (as low as −63.95‰) suggesting a possible relationship to sulfate reducing bacteria, which is common during anaerobic oxidation of methane at seep sites.