A model describing the time course and variability in toxicity of hydrophobic chemicals to aquatic organisms

The growth of Chlorella marine, Nannochloris oculate, Pyramimonaos sp., Platymonas subcordiformis and Phaeodactylum tricornutum exposed to chlorobenzene, 1,2-dichlorobenzene, 1,2,3,4-tetrachlorobenzene and pentachlorobenzene was tested. The Boltzman equation was used to describe organism growth. The time course for uptake of hydrophobic organic chemicals (HOCs) by aquatic organisms was expressed by incorporating growth and, if desired, the effect of metabolism into the HOC bioconcentration process. The probability of any given concentration of HOCs in the organisms causing a specified toxic endpoint was expressed with a modified Weibull distribution function. The combined bioconcentration and probability equations were tested with data for time course of incubation of algae exposed to chlorinated benzenes (CBs). A set of parameters, including the uptake rate constant k ₁, the elimination rate constant k ₂ and thereafter the bioconcentration factor on a dry weight basis, BCF _D, the critical HOC concentration in the organism resulting in a specified toxic endpoint, C _A ^* , and the spread factor, S, could be obtained by fitting only experimental data for percent growth inhibition(%)-time-CB exposure concentration. The average coefficients of variation within CBs were 15.2% for BCF _D, 21.0% for k ₁, 18.3% for k ₂, 8.1% for C _A ^* and 9.7% for S. The variability in toxicity (such as EC₁₀, EC₅₀, EC₉₀) derived from the model equations agreed well with those experimentally observed.     

Heterotrophic bacterial flora in aquaculture area around Xuejiadao

From Oct., 1999 to Oct., 2000, the heterotrophic bacterial flora in the aquaculture area around Xuejiadao was investigated. The result shows that the populations of the heterotrophic bacteria are heavier in summer and autumn than those in winter and spring. The average populations in seawater, sediment, the surface of seaweed and the surface of fish are 1.4×10⁴cfu mL⁻¹, 5.4×10⁶cfu g⁻¹, 1.5×10⁶cfu g⁻¹ and 1.8×10³cfu cm⁻², respectively. A total of 301 strains were isolated, among them 259 were Gram-negative. All the Gram-negative bacteria belong to 13 genera and some genera of Enterobacteriaceae. The communities of bacteria are slightly different among the samples. In the body surface of fish, Genus vibrio is dominant. In the remaining samples, dominant genus is Aeromonas.     

Life cycle and production of chironomidae (diptera) in Biandantang, a typical macrophytic lake (Hubei, China)

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Bioflocculant Produced by Klebsiella sp. MYC and Its Application in the li＇eatment of Oil-field Produced Water

Seventy-nine strains of bioflocculant-producing bacteria were isolated from 3 activated sludge samples. Among them, strain MYC was found to have the highest and stable flocculating rate for both kaolin clay suspension and oil-field produced water. The bacterial strain was identified as Klebsiella sp. MYC according to its morphological and biochemical characteristics and 16SrDNA sequence. The optimal medium for bioflocculant production by this bacterial strain was composed of cane sugar 20 g L^-1, KH2PO4 2 g L^-1, K2HPO4 5 g L^- 1, （ NH4）2SO4 0.2 g L^-1, urea 0.5 g L^- 1 and yeast extract 0.5 g L^- 1, the initial pH being 5.5. When the suspension of kaolin clay was treated with0.5% of Klebsiella sp. MYC culture broth, the flocculating rate reached more than 90.0% in the presence of 500mg L^-1 CaCI2, while the flocculating rate for oil-field produced water was near 80.0% in a pH range of 7.0 - 9.0 with the separation of oil and suspended particles from the oil-field produced water under similar conditions. The environment-friendly nature of the bioflocculant and high flocculating rate of the strain make the bioflocculant produced by Klebsiella sp. MYC an attractive bioflocculant in oil-field produced water treatment.     

Species composition and gear characteristics of the <Emphasis Type="Italic">Macrobrachium</Emphasis> fishery of the Cross River Estuary,Nigeria

The Cross River Estuary, Nigeria, is an important shrimping area for artisanal fishermen of the coastal communities. The multi-species Macrobrachium fishery is exploited with three main gears, namely beach seine, push net and trap. Studies on species composition of this fishery recorded thirteen shrimp species, one swimming crab (Callinectes amnicola) and two fish species (Eleotris sp. and Pellonula leonensis). The shrimp species identified included Macrobrachium macrobrachion (83.39% and 55.69% by number and weight, respectively), M. vollenhovenii (9.66% and 37.18%), M. equidens (3.8% and 2.87%), juveniles-sub-adults of Penaeus notialis (1.11% and 1.3%), M. dux, M. felicinum, Palaemonetes africanus, Palaemon maculatus, Palaemon elegans, Desmocaris sp., Leander sp., Nematopalaemon hastatus and Alpheus pontederiae. While the selectivity index for trap was 0.25, beach seine and push net had a lower index of 0.063. The results present the first comprehensive and representative report for the Estuary shrimp fishery and will assist in the management of the biodiversity of this ecosystem.     

ORGANIC GEOCHEMISTRY MARKERSOF CLIMATIC EVOLUTION IN LOESSREGION OF CHINA

Two hundred and thirty-five samples collected from Weinan loess section (L₂ – S₀) have been analyzed by TOC (total organic carbon) and pyrolysis hydrocarbons. The curve of TOC with depth has been set up and compared well with other climatic markers. The main periods of TOC variation of the last 0.14 Ma are 67.08 ka and 18.29 ka, which is near with the periods of the variation of sunshine, showing that the TOC variation may be controlled by the variation of 65°N sun-shine. The distribution of pyrolysis hydrocarbon and its parameters shows that there are two kinds of hydrocarbon associated with climatic change. One is called hydrocarbon pyrolysted directly from organism (pHC_2–1) which is in the range of T _max350°C – 500°C, and associated with atmospheric water. The other is called carbonate inclusion hydrocarbon (pHC_2–2) which is in the range of T _max 500°C – 600°C and formed by evaporation and associated with dry climate. The formula calculating humidity has been deduced Og=pHC_2–1/(pHC_2–1 + pHC_2–2). The range of humidity of the section is 47% – 77% and the reducing order is S_1–3, S_1–2, S₀, S_1–1, S_1–4, S_1–1, L_1–1, L_1–3, and L_1–5. And the trend of the humidity is still increasing today.     

PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp. on intestinal microbial diversity in kuruma shrimp (Marsupenaeus japonicus)

In this study, the intestinal microbiota of kuruma shrimp (Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects of Bacillus spp. on intestinal microbial diversity. Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp. amended feed. PCR and denaturing gradient gel electrophoresis (DGGE) analyses were then performed on DNA extracted directly from the guts. Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S rDNA amplicons, and distinct bands in the gels were sequenced. The results suggested that the gut of kuruma shrimp was dominated by Vibrio sp. and uncultured gamma proteobacterium. Overall, the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity of shrimp.     

A NEW SPECIES OF THE GENUS NEROCILA （ISOPODA： CYMOTHOIDAE） FROM THE EAST CHINA SEA

A new species of cymothoid Isopeda, Nerocila donghaiensis, collected from the body of a sciaenid fish, Johnius sp., is described in the present paper. The new species can be distinguished from the other species of genus Nerocila by its endoped of uropods with deep notch, sharply serrated on the lateral margin and strongly serrated on the medial margin; antennule with 1 st article swollen; dactyli of pereopods Ⅰ, Ⅱ, Ⅳ, Ⅴ swollen.     

Isolation and characterization of phosphate-solubilizing bacteria from seagrass rhizosphere soil

Phosphate-solubilizing bacterial strains(6 Nos.) were isolated from the rhizosphere soils of two seagrasses(Halophila ovalis(R.Br.) Hook and Halodule pinifolia(Miki) Hartog) in the Vellar estuary.Experimental studies found that the strain PSSG6 was effective in phosphate solubilization with Phosphate Solubilization efficiency index E = 375 ± 8.54,followed by the strain PSSG5 with Phosphate Solubilization efficiency index E = 275 ± 27.3.Of the 6 strains isolated,the strains PSSG4 and PSSG5 be-longed to the genus Bacillus,and PSSG1,PSSG2 and PSSG3 were identified as Citrobacter sp.,Shigella sp.,and Klebsiella sp.,respectively,by conventional method,and PSSG6 was identified as Bacillus circulans using conventional and molecular methods.     

Heterotrophic bacterial floras in industrial area and unpolluted marine environments around Qingdao

From Oct. 1999 to Oct. 2000, the heterotrophic bacterial floras in the industrial marine environment around the Qingdao Power Plant (QPP) and in the unpolluted marine environments were investigated. The results showed that the numbers of the heterotrophic bacteria around QPP were much higher than those in unpolluted environments, and the average numbers in QPP Seawater, QPP Sediment, Unpolluted Seawater and Unpolluted Sediment were 5.4×10⁴cfu(mL)⁻¹, 5.0×10⁵cfug⁻¹, 3.0×10²cfu(mL)⁻¹ and 1.3×10⁵cfug⁻¹ respectively. Totally, 118 strains were isolated from QPP and 99 of them were Gram-negative. One hundred and twenty one strains were isolated from the unpolluted environments and 104 of them were Gram-negative. All the Gram-negative bacteria belonged to 13 genera. The distribution of the bacteria was varied in different marine environments. The results showed that the unpolluted marine environments contained much more Vibrio than seawater and sediment around QPP.     

Biodegradation of <Emphasis Type="Italic">Enteromorpha</Emphasis> polysaccharides by intestinal micro-community from <Emphasis Type="Italic">Siganus oramin</Emphasis>

Micro-communities are supposed to have more potential functions of biodegradation of polysaccharides than single strain; however, the intestinal micro-communities involved in the biodegradation of Enteromorpha polysaccharides (EP) were seldom reported. In order to obtain the EP-degrading micro-community, the intestines of Siganus oramin was obtained to isolate the micro-communities, which were enriched by 0.3% of EP as the sole carbon source. A stable micro-community with EP degradative capability was achieved after seven generations of subculture, named H1. Results showed that H1 was able to degrade 75% of EP within 24 hours, and the activity of EP lyases reached 500 U mL^?1 in 32 hours. With denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analysis, ten bacteria closely related to Marinomonas pontica, Microbacterium sp., Leucobacter chironomi, Cyclobacterium sp., Algoriphagus winogradskyi, Pseudoalteromonas sp. and Vibrio sp. were determined. Furthermore, compared with the DGGE bands sequence and the clone library analysis, the dominant bacteria of the EP-biodegrading micro- community were Pseudoalteromonas sp. and Vibrio sp., with the respective proportion of 38% and 46%, and they should play an important role in EP degradation together with other degrading bacteria in the micro-community H1.     

Isolation,purification and characterization of the hydrogen evolution promoting factor of hydrogenase ofSpirulina platensis

A component (s-factor) with obvious promoting effect on hydrogen evolution of hydrogenase has been isolated and extracted from a cell-free preparation ofSpirulina platensis. The effect of the s-factor in the reaction system is similar to that of Na₂S₂O₄, but is coupled with light. The s-factor has the maximum absorption peak at 620 nm in the oxidized state, at 590 nm in the reduced state. The partially purified s-factor showed two bands by SDS-PAGE and is distinctly different from phycocyanin, which has no change of oxidized state and reduced state absorption spectra, and also has no promoting effect on hydrogenase ofSpirulina platensis under the light. The research supported by the NSFC.     

Preliminary identification of three new isolates in genus Alexandrium (Dinophyceae) from China sea area

The 5.8S ribosomal DNA sequences (5.8S rDNA) and their flanking regions, internal transcribed spacer 1 and spacer 2 (ITS1 and ITS2) of three new isolates in genus Alexandrium (Alexandrium sp. qd1, Alexandrium sp. qd2, Alexandrium sp. gz) from China were amplified, sequenced, and subjected to phylogenetic analysis. Alexandrium sp. gz and Alexandrium sp. qd1 were grouped with high bootstrap values with four strains/species, i.e., A. catenella South Korea strain, A. catenella Japan strain, and two from China, Alexandrium sp. AC03 and Alexandrium sp. AN01 being proposed to be A. catenalla in a previous study. Then Alexandrium sp. gz and Alexandrium sp. qd1 were identified as Alexandrium catenella. As A. catenella was isolated from Qingdao and Guangzhou sea areas, it supposedly distributed at least in these two areas and was genetically different. Alexandrium sp. qd2 differed greatly from species in Alexandrium. It clustered with Symbiodinium californium, Symbiodinium sp. G15 and Gymnodinium sp. Zhao 01 with 100% bootstrap value; so Alexandrium sp. qd2 affiliates to genus Symbiodinium, and is probably a free-living Symbiodinium species.     

Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification

Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification (LAMP) assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase (empA) gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65°C in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V. anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.     

Sequence polymorphism of two major histocompatibility (MH) class II B genes and their association with Vibrio anguillarum infection in half-smooth tongue sole (Cynoglossus semilaevis)

Major histocompatibility complex (MHC) class Ⅱ B molecules play an important role in the adaptive immune response in fish. Previous study has reported that two highly polymorphic class ⅡB genes, Cyse-DAB and Cyse-DBB exist in half-smooth tongue sole (Cynoglossus semilaevis). In this study, the polymorphism within exon 2 of the class Ⅱ B genes following bacterial challenge was evaluated. Two hundred C. semilaevis individuals were injected intraperitoneally with Vibrio anguillarum. Muscle tissue from the first 20 dead and 20 of the survivors was collected for genotyping. Sixty alleles from the 40 individuals were isolated, of which 32 belonged to Cyse-DAB and 28 belonged to Cyse-DBB. The rate of dN (non-synonymous substitution) was higher than that of dS (synonymous substitution) in the PBRs (peptide binding residues) of both class Ⅱ B genes. Conversely, the rate of dS was higher than dN in the non-PBRs and the complete exon 2 sequence. Thus, the results suggest that positive selection has occurred in the PBRs and purifying selection in the non-PBRs and exon 2. Thirteen class Ⅱ B alleles were used to study the association between alleles and resistance to infection. Though not significant, alleles Cyse-DAB*0601, Cyse-DAB*0706, and Cyse-DBB*0101, Cyse-DBB*1301 were only found in surviving individuals and may represent alleles that have resistance against V. anguillarum infection. Alleles Cyse-DAB*0701 and Cyse-DAB*1301 were significantly more prevalent in dead individuals than in surviving ones and may represent alleles that are associated with increased susceptibility to V. anguillarum infection.     

Soil organic carbon pools in particle-size fractions as affected by slope gradient and land use change in hilly regions, western Iran

This study was conducted to explore the effects of topography and land use changes on particulate organic carbon(POC),particulate total nitrogen(PTN),organic carbon(OC) and total nitrogen(TN) associated with different size primary particle fractions in hilly regions of western Iran.Three popular land uses in the selected site including natural forest(NF),disturbed forest(DF) and cultivated land(CL) and three slope gradients(0-10 %,S1,10-30 %,S2,and 30-50%,S3) were employed as the basis of soil sampling.A total of 99 soil samples were taken from the 0-10 cm surface layer in the whole studied hilly region studied.The results showed that the POC in the forest land use in all slope gradients was considerably more than the deforested and cultivated lands and the highest value was observed at NF-S1 treatment with 9.13%.The values of PTN were significantly higher in the forest land use and in the down slopes(0.5%) than in the deforested and cultivated counterparts and steep slopes(0.09%) except for the CL land use.The C:N ratios in POC fraction were around 17-18 in the forest land and around 23 in the cultivated land.In forest land,the silt-associated OC was highest among the primary particles.The enrichment factor of SOC,EC,was the highest for POC.For the primary particles,EC of both primary fractions of silt and clay showed following trend for selected land uses and slope gradients:CL> DF> NF and S3 > S2> S1.Slope gradient of landscape significantly affected the OC and TN contents associated with the silt and clay particles,whereas higher OC and TN contents were observed in lower positions and the lowest value was measured in the steep slopes.Overall,the results showed that native forest land improves soil organic carbon storage and can reduce the carbon emission and soil erosion especially in the mountainous regions with high rainfall in west of Iran.     

Nemerteans of Zhoushan Islands,East China Sea coast,China II. Enopla: Distromatorhynchocoela: Monostilifera: Tetrastemmidae

A new monostiliferous hoplonemertean,Psammamphiporus microocelli sp. nov., is described. It differs from the single species of genusPsammamphiporus, P. elongatus, in being without a frontal organ and cephalic furrows, the stylet basis being conical rather than cylindrical. The project supported by National Natural Science Foundation of China.     

Biomass growth characteristics of 13-year-old <Emphasis Type="Italic">Pinus densiflora</Emphasis> S. et Z. in a post-fire plantation on different contour conditions in Samcheuk,Korea

We used preliminary data to estimate the growth volume of artificially reforested Pinus densiflora in a post-fire area on three different contour conditions. We compared the growth of P. densiflora on a south-facing slope (S_sth), north-facing slope (S_nth) and ridge area (Ridge), using 7 trees selected from each stand aspect. The tree height, diameter and growth volume were measured and the dry weight of each plant part were compared and analyzed. The results revealed that the total dry weight was highest on S_sth (5992.3 g), followed by S_nth (4833.2 g) and lowest on Ridge (3160.1 g). The height growth was highest on S_nth (285.8 cm), followed by S_sth (274.5 cm) and lowest on Ridge (211.5 cm). The diameter growth was greatest on S_sth (7.37 cm), followed by S_nth (7.10 cm) and lowest on Ridge (5.72 cm). The volume growth was highest on S_sth (4257.7 cm³), followed by S_nth (3750.7 cm³) and lowest on Ridge (2093.7 cm³). Therefore, we should consider and include the concept of slope orientation together with differences in habitat environments in afforestation projects when creating artificial forests with P. densiflora. These study results can serve as important preliminary data for future establishment of artificial forest of P. densiflora in a post-fire plantation.     

Effects of heavy metals （Pb^2＋ and Cd^2＋） on the ultrastructure, growth and pigment contents of the unicellular cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb²⁺ and Cd²⁺ (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (>4 mg/L Pb²⁺) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L Pb²⁺), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L Pb²⁺), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll α, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb²⁺, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll α, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd²⁺concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC₅₀) values (3.47 mg/L Cd²⁺ and 12.11 mg/L Pb²⁺) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd²⁺ than Pb²⁺. Supported by the Chinese Scholarship Council     

Global ocean tides from Geosat altimetry by Quasi-Harmonic analysis

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