Recover vigorous cells of Magnetospirillum magneticum AMB-1 by capillary magnetic separation

Cultivable magnetotactic bacteria (MTB) in laboratory can provide sufficient samples for molecular microbiological and magnetic studies. However, a cold-stored MTB strain, such as Magnetospirillum magneticum AMB-1, often loses its ability to synthesize magnetosomes and consequently fails to sense the external magnetic field. It is therefore important to quickly recover vigorous bacteria cells that highly capable of magnetosome producing. In this study, a modified capillary magnetic separation system was designed to recover a deteriorating strain of Magnetospirillum magneticum AMB-1 that long-term cold-stored in a refrigerator. The results show that all cells obtained after a 3-cycle treatment were vigorous and had the ability to produce magnetosomes. Moreover, the 3^rd-cycle recovered cells were able to form more magnetosome crystals. Compared with the colony formation method, this new method is time-saving, easily operated, and more efficient for recovering vigorous MTB cells.     

Lytic Characteristics and Identification of Two Alga-lysing Bacterial Strains

Lytic Characteristics and Identification of Two Alga-lysing Bacterial StrainsBeaulieu, S. E., M. R. Sengco, and D. M. Anderson, 2005. Using clay to control harmful algal blooms: deposition and resuspension of clay/algal flocs. Harmful Algae . (4): 123-138…     

Ultrastructural observations on sporogenesis ofMyxobolus Kwangsiensis Hsieh,Gong & Xiao from the kidney of daceLabeo rohita

The early stages and capsulogenetic stages of sporogenesis ofMyxobolus kwangsiensis were studied in detail by electron microscopy.M. kwangsiensis is an intracellular myxosporean infecting the epithelial cells of kidney tubules of young daceLabeo rohita. TheM. kwangsiensis sporogensis presents the typical features of myxosporea. The initial stages are multinucleate primary cells, with one primary cell typically enclosing one secondary cell. The secondary cells are formed directly by cytokinesis inside the primary cells. The primary cell participates in the formation of the secondary cell nucleus. The nuclear division of primary cell is asynchronous by amitosis. The parasitic organisms may degenerate inside the epithelial cells at any stage. The modes of sporoblast formation, the typical features of myxosporea and the cause of its degradation are discussed. This research was supported by the NSFC. (No. 39070675)     

Biofilm formation under high temperature causes the commensal bacteria Bacillus cereus WPySW2 to shift from friend to foe in Neoporphyra haitanensis in vitro model

Although biofilm formation may promote growth, biofilms are not always beneficial to their hosts. The biofilm formation characteristics of Bacillus cereus WPySW2 and its changes at different temperatures were studied. Results show that B. cereus WPySW2 promoted the growth of Neoporphyra haitanensis (an economically cultivated seaweed) at 20 °C but accelerated algal rot at 28 °C. Thicker B. cereus WPySW2 biofilms covered the surface of N. haitanensis thalli at 28 °C, which hindered material exchange between the algae and surrounding environment, inhibited algal photosynthesis and respiration, and accelerated algal decay. Compared with planktonic bacteria, mature biofilm cells had lower energy consumption and metabolic levels. The biofilm metabolic characteristics of B. cereus WPySW2 changed significantly with temperature. High temperature accelerated biofilm maturation, which made it thicker and more stable, allowing the bacteria to easily adapt to environmental changes and obtain greater benefits from their host. High temperature did not affect the production or increased the abundance of toxic metabolites, indicating that the negative effects of B. cereus WPySW2 on algae were not caused by toxins. This study shows that increased temperature can transform a harmless bacterium into a detrimental one, demonstrating that temperature may change the ecological function of phycospheric bacteria by affecting their morphology and metabolism.

     

Electrochemical properties of electrodes with different shapes and diffusion kinetic analysis of microbial fuel cells on ocean floor

Microbial fuel cell(MFC) on the ocean floor is a kind of novel energy-harvesting device that can be developed to drive small instruments to work continuously.The shape of electrode has a great effect on the performance of the MFC.In this paper,several shapes of electrode and cell structure were designed,and their performance in MFC were compared in pairs:Mesh(cell-1) vs.flat plate(cell-2),branch(cell-3) vs.cylinder(cell-4),and forest(cell-5) vs.disk(cell-6) FC.Our results showed that the maximum power densities were 16.50,14.20,19.30,15.00,14.64,and 9.95 mWm-2 for cell-1,2,3,4,5 and 6 respectively.And the corre-sponding diffusion-limited currents were 7.16,2.80,18.86,10.50,18.00,and 6.900 mA.The mesh and branch anodes showed higher power densities and much higher diffusion-limited currents than the flat plate and the cylinder anodes respectively due to the low diffusion hindrance with the former anodes.The forest cathode improved by 47% of the power density and by 161% of diffusion-limited current than the disk cathode due to the former’s extended solid/liquid/gas three-phase boundary.These results indicated that the shape of electrode is a major parameter that determining the diffusion-limited current of an MFC,and the differences in the elec-trode shape lead to the differences in cell performance.These results would be useful for MFC structure design in practical applica-tions.     

Toxic effects of Karenia mikimotoi extracts on mammalian cells

Karenia is one of the most harmful and representative red tide genus in a temperate zone. Blooms caused by this genus have resulted in massive fish death in the South China Sea and the East China Sea. However, the potential effects of this dinoflagellate on human health through the transfer of toxins via marine food webs, and the mechanisms of toxicity, are still unknown. Therefore, we examined the toxic effects of a strain of K. mikimotoi (isolated from the South China Sea) on the proliferation and morphology of four mammalian cell lines (two normal cell lines and two cancer cell lines). In addition, we carried out a preliminary investigation on the mechanism of toxicity of the alga. The results show that the polar lipid-soluble component of K. mikimotoi significantly inhibited proliferation of the four cell lines, and resulted in the cells becoming spherical, swollen and damaged. The result of Annexin V and PI double-staining confirmed that cell membranes were disrupted. The malonaldehyde (MDA) contents in the medium of the four cell lines treated with the polar-lipid extracts all increased significantly, which indicates that the polar-lipid toxins produced by K. mikimotoi could adversely affect mammalian cells by inducing lipid peroxidation. We conclude that K. mikimotoi is a potential threat to human health, and the comprehensive effect of this dinoflagellate and its mechanisms should be investigated further.     

On a new species ofPavlova (Prymnesiophyceae) from China

Pavlova viridis sp. nov. is described on the basis of light and electron microscope observations. The material was collected from the coast of haiyang county of Shandong, China. It is characterized by 1) yellowish green to green chloroplast, 2) rudimentary short flagellum 0.3 μm long and hook-shaped, 3) long flagellum with small spherical knob-scales, 4) absence of pyrenoid and stigma, and 5) marine habitat. This new species andP. salina seem to be most closely related to each other but their colour, the insertion of their three appendages, the shape of their knob scales, and their cell periplasts are quite different from one another. Contribution No. 1879 from the Institute of Oceanology, Academia Sinica     

Adhesion mechanisms of Vibrio fluvialis to skin mucus of Epinephelus awoara

Vibrio fluvialis incubated in trypticase soy broth (TSB) showed stronger adhesion to the skin mucus of Epinephelus awoara than V. fluvialis grown on trypticase soy agar (TSA), and this bacterial adhesion was assessed in terms of saturation kinetics. Treating bacteria with antibody against O-antigens resulted in significantly reduced bacterial adhesion. In the early growth stage, the adhering bacteria numbers increased with incubation time, peaked at 24 h, and then dropped sharply. Prior heat treatment of the mucus at various temperatures resulted in different effects on bacterial adhesion. No remarkable effect on bacterial adhesion was detected after treating mucus with protease K and trypsin, whereas periodic acid treatment significantly enhanced bacterial adhesion. Extracts of V. fluvialis surface structures at high concentrations significantly inhibited bacterial adhesion, while relatively low concentrations enhanced adhesion. V. fluvialis had strong affinity for a high molecular weight mucus component. These results indicate that V. fluvialis adhered strongly to E. awoara mucus by specific adhesion and that this adhesion was influenced by many factors, including bacterial conditions, mucus, and the environment.     

Characterization of Dust and Non-dust Aerosols with SEM/EDX

Total suspended particulate （TSP） samples were collected at three sites along the coast of Qingdao, China, before and during a major dust storm in March, 2002. For comparison, PM10（particulate matter with aerodynamic diameters less than 10μm） samples were collected at one of the three sites. The morphological observation and compositional analysis of bulk and individual particles were performed by using scanning electron microscopy equipped with an energy dispersive X-ray system （SEM/EDX） for the TSP and PM10 samples. The results showed that the particles of different kinds of morphology had different elemental compositions, but the particles of similar morphology did not always have the same elemental composition for non-dust samples. The morphology and composition of non-dust particles were different at different sites. The fractal and spherical particles existed mainly in the coarse fraction for non-dust samples, while in the fine mode （〈 10μm） there were floccules formed by fine particles flocking together and containing crustal elements. Compared with the non-dust particles, the dust particles were more homogeneous in terms of morphology, particle size and composition. Particles with irregular shapes and well-distributed sizes dominated in the dust samples, containing crustal elements such as Mg, A1, Si, Ca, Fe, etc. The high sulfur content indicated that homogeneous and heterogeneous reactions took place on the surfaces of the dust particles in the specific environment of Qingdao.     

Culture techniques and growth characteristics of Dinophysis acuminata and its prey

The dinoflagellate Dinophysis acuminata Claparede & Lachmann is a toxic alga that causes diarrhetic shellfish poisoning. No Dinophysis species were maintained in culture for a long period of time until 2006 when Park successfully established D. acuminata in culture using a three-step feeding protocol in which the cryptophyte, Geminigera cryophila, is fed to Myrionecta rubra (=Mesodinium rubrum), a ciliate that is in turn fed to D. acuminata. In this paper, we present the details of culturing D. acuminata from the Northeastern United States. The protocols described herein can be adopted for laboratory studies of this species. The effects of temperature on the growth and ingestion rates of D. acuminata were also examined. The results show that D. acuminata growth rate was 0.23 /d at 10°C and 0.11 /d at 4°C when fed M. rubra prey. The maximum prey ingestion rate was 2.80 Dinophysis cell/d at 10°C, although the rate decreased slightly at 4°C. In overall, temperature showed a greater influence on growth rate of D. acuminata than on the ingestion rate under the study conditions, and the quantity of available food was also an important regulator to D. acuminata growth.     

Notes on Two Marine Ciliates from the Yellow Sea,China： Placus salinus and Strombidium apolatum（Protozoa, Ciliophora）

The living morphology and infraciliature of two rare marine ciliates, Placus salinus Dietz, 1964 and Strombidium apolatum Wilbert and Song, 2005, collected from the coastal waters near Qingdao, China, were investigated by in vivo observation and protargol impregnation technique. The improved diagnosis for Placus salinus is as follows: medium-sized marine Placus, in vivo (50–60)μm×(30–40)μm; cell elliptical to barrel-shaped; 28–31 somatic kineties; single macronucleus usually ellipsoid and one micronucleus located in the indention of the macronucleus; one contractile vacuole posteriorly positioned. Strombidium apolatum is characterized by: marine strombidium (40–60)μm×(30–45)μmm in vivo, cordiform in shape with somewhat pointed posterior end and conspicuous apical protrusion; extrusomes prominent, about 15μm in length and evenly arranged along the circle kinety; about 16 collar and 5–6 buccal membranelles; one elongate macronucleus and one micronucleus; circle and ventral kineties consisting of about 53 and 45 dikinetids respectively.     

A VIBRIO ANGUILIARUM STRAIN ASSOCIATED WITH SKIN ULCER ON CULTURED FLOUNDER, PARALICHTHYS OLIVACEUS

The characteristics of a bacterium strain M3, isolated from cultured flounderParalichthys olivaceus with remarkable external sign of skin ulcer during an epizootic outbreak, indicated that the bacterium belonged to the speciesVibrio anguillarum. Challenge by I.M. (intramuscular injection), bath, and oral administration with M3 showed that it was highly pathogenic forParalichthys olivacues. The LD₅₀ dose was 5.144×10³ CFU/ per fish infection by I.M. injection. Recovered inoculated bacteria from the surviving fish revealed that the asymptomatic carriers could be a latent contagious source. Study of the effect of bacterial culture CFS (cell-free-supernatant) showed that the exotoxins produced by M3 play an important role in its pathogenicity for flounder. The resistance of M3 to 36 out of 41 antibiotics indicated that the bacterial disease outbreak was mainly attributable to the frequent and excessive use of antimicrobial agents; and that vaccination would be an effective precaution against bacterial disease. Contribution No. 4107 from Institute of Oceanology, Chinese Academy of Science. Contribution No. 270 from Experimental marine Biology Laboratory, CAS. Funded by projects under the Major State Basic Research Development Program, Grant G1999012003.     

Winter Phytoplankton Assemblages of Coastal Yellow Sea Connected to Jiaozhou Bay,China

I Introduction Phytoplankton play an important role in the primary production of ocean (Ning et al., 1995). They are impor-tant biological mediators of carbon turnover in seawater ecosystems (Zhu et al., 1993). Phytoplankton in Jiaozhou Bay have been preliminarily studied on the subjects of community structure, primary productivity and carbon budget (Qian et al., 1983; Guo et al., 1992; Jiao et al., 1994). It has been found that seasonal variation of phytoplankton cell abundance presents w…     

Effects of Microcystis aeruginosa on life history of water flea Daphnia magna

Cyanobacterial blooms in eutrophic freshwater systems are a worldwide problem, creating adverse effects for many aquatic organisms by producing toxic microcystins and deteriorating water quality. In this study, microcystins (MCs) in Microcystis aeruginosa, and Daphnia magna exposed to M. aeruginosa, were analyzed by HPLC-MS, and the effects of M. aeruginosa on D. magna were investigated. When D. magna was exposed to M. aeruginosa for more than 2 h, Microcystin-LR (MC-LR) was detected. When exposed to 1.5 × 106, 3 × 106, 0.75 × 107, and 1.5 × 107 cell/mL of M. aeruginosa for 96 h, average survival of D. magna for treatments were 23.33%, 33.33%, 13.33%, 16.67%, respectively, which were significantly lower than the average 100% survival in the control group (P < 0.05). The adverse effects of M. aeruginosa on body length, time for the first brood, brood numbers, gross fecundity, lifespan, and population growth of D. magna were density-dependent. These results suggest that the occurrence of M. aeruginosa blooms could strongly inhibit the population growth of D. magna through depression of survival, individual growth and gross fecundity. In the most serious situations, M. aeruginosa blooms could undermine the food web by eliminating filter-feeding zooplankton, which would destroy the ecological balance of aquaculture water bodies.     

Establishment of a spleen cell line from large yellow croaker Pseudosciaena crocea and its primitive application in foreign gene transfection

A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco’s modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies.     

Effect of temperature and irradiance on the growth and reproduction of Enteromorpha prolifera J. Ag. （Chlorophycophyta,Chlorophyceae）

Effect of temperature and irradiance on growth and reproduction of Enteromorpha prolifera that bloomed offshore along the Qingdao coast in summer 2008, was studied. It was showed that E. prolifera propagated mainly asexually with specific growth rate （SGR） of 10.47 at 25℃/40 μmol m^-2s^-1. Under this condition, gametes with two flagellate formed and released in 5 days. At the beginning of the development, the unicell gamete divided into two cells with heteropolarity, and then the apical cell developed into thalli primordial cells, whereas the basal cell developed into rhizoid primordial cells. In 8-day culture, the monoplast gamete developed into juvenile germling of 240 μm in length. Unreleased gametes can develop directly within the alga body. E. prolifera could either reproduce through lateral branching or fragmenting except apomixis revealed by Microscopic observation. On aged tissue of E. prolifera, although the degraded pigments partially remained in faded algal filaments, numerous vegetative cells could still divide actively in the algal tissues.     

Variations in retention efficiency of bivalves to different concentrations and organic content of suspended particles

Retention efficiencies (RE) of scallop (Chlamys farreri), oyster (Crassostrea gigas) and mussel (Mytilus edulis) in a flow-through system were measured to understand the short-term response to various particle and organic matter concentrations. By comparing the RE of C. farreri with that of C. gigas and M. edulis, we gained further knowledge on the feeding physiological characteristics of C. farreri and ascertained the possible cause of high summer mortalities of this species. The experimental feeding conditions included natural differences in the abundance and composition of suspended seston, as well as conditions in which seston abundance and composition were manipulated by adding natural silt or cultured microalgae. The results show that in natural sea water, the minimum particle size for maximal retention in M. edulis, C. gigas and C. farreri was approximately 4, 6, and 8 μm, respectively; the RE of 2-μm (equivalent spherical diameter) particles was 17%, 19%, and 8%, respectively; and the relative standardized RE was 58%, 49%, and 18%, respectively. In C. gigas and M. edulis, the minimal particle size for maximal retention did not change with food quality (organic content). C. farreri was more sensitive to fluctuations in particle concentration and organic content. With particle concentration increase, the minimal particle size for maximal retention in scallop shifted to large particles and the RE for 2-μm particle decreased from 8% in natural seawater to 1.6%–6% in silt-enriched groups. With organic content increase, the minimal particle size for maximal retention shifted from 8 to 5 μm in natural seawater.Variation in RE of C. farreri with food conditions and the relationship between lower RE and smaller particle size may hamper C. farreri from food taking due to the decrease in the size distribution of phytoplankton in Sungo Bay.     

Early stage differentiation of thallus cells ofPorphyra haitanensis (Rhodophyta)

The early stage differentiation of thallus cells ofPorphyra haitanensis T. J. Chang et B. F. Zheng was studied. Protoplasts or single cells were isolated from the blades using enzyme mixture comprising 2% sea snail gut enzyme and 1% cellulase. The isolated protoplasts or single cells were incubated in the MES medium. The cell differentiations were examined under the microscope at intervals after incubation. Four types of cell differentiation, namely, normal, abnormal, carposporangial and spermatorangial, and rhizoidal types, were observed. Since normal cell differentiations occur mostly in small thalli 50 mm in length and middle portions of big thalli 200 mm in length, it is essential to select tissues from these two kinds of thalli essential for commercial production.