Kinetics of alkaline phosphatase in lake sediment associated with cage culture of Oreochromis niloticus

Variations in kinetics of alkaline phosphatase occurring in different sites of sediment associated with cage culture of Oreochromis niloticus in a shallow Chinese freshwater lake (Lake Donghu) were described. In addition, the kinetic parameters of each 2.5-cm stratum in the sediment from the surface down to 37.5 cm were analyzed. Horizontally, the V-max values of alkaline phosphatase in surface sediments increased markedly at sites immediately under and adjacent to the cage that would be subjected to the deposition of fish feces. Peak V-max values in the top 5 cm of the sediment under the cage were also observed relative to their deeper control. After a treatment where the fish feces were added over 12 days, the sediment in deeper layer exhibited a significantly higher V-max value, thereby corroborating the relationship between V-max values of alkaline phosphatase and fish feces in sediments. The fish feces exhibited a remarkable alkaline phosphatase activity (APA). Thus, it is indeed a source of the enzyme. Effects of the fish feces were dose- and time-dependent. The V-max values in sediments were always stimulated, but the K-m values showed much more variability. (C) 2001 Elsevier Science B.V. All rights reserved.

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.

Changes in culture growth dynamics and cell size in a Scenedesmus quadricauda upon action of potassium dichromate

The effect of potassium dichromate in concentrations of 0.5 to 10 mg/l on a laboratory culture of Sc. quadricauda algae was studied in standard conditions. The total cell numbers decreased at potassium dichromate concentrations over 1 mg/l, and the proportion of living cells decreased at all studied concentrations. A positive correlation was found between changes in cell size and their numbers at toxin concentrations of 1 and 3 mg/l, and a negative correlation was found between the relative size and the cell numbers at 3 and 10 mg/l. This may be due to different intensity of growth inhibition and cell division under the influence of the toxin. The culture sensitivity to the toxin increased in autumn and decreased in the spring.

FATE OF 2 PHENYLBENZOYLUREA INSECTICIDES IN AN ALGAE CULTURE SYSTEM (SCENEDESMUS-SUBSPICATUS)

The bioaccumulation, elimination and degradation of C-14-labelled diflubenzuron (DFB) and of 1-(2-chlorobenzoyl)-3-(4-chlorophenyl)urea (CCU) was studied in a laboratory algae culture system of scenedesmus subspicatus. Algae were exposed at an initial concentration of 200 mug/l for seven days. Neither substance had an inhibitory effect on the growth of algae. The half life of DFB and CCU was 3 and 1 days, respectively, as measured by HPLC. The distribution of C-14 between medium and algae was measured. In the case of DFB radioactivity in algae increased steadily and levelled off at approximately 60 % after 5 days. Due to algae growth BCF values decreased from 4310 to 889 for DFB and from 6719 to 304 for CCU during the exposure period. The relationship between algae density and bioconcentration could be correlated by an adsorption isotherm. Elimination of both compounds was rapid during the first hours.

Hydrogen production characteristics of the organic fraction of municipal solid wastes by anaerobic mixed culture fermentation

The hydrogen production from the organic fraction of municipal solid waste (OFMSW) by anaerobic mixed culture fermentation was investigated using batch experiments at 37 degrees C. Seven varieties of typical individual components of OFMSW including rice, potato, lettuce, lean meat, oil, fat and banyan leaves were selected to estimate the hydrogen production potential. Experimental results showed that the boiling treated anaerobic sludge was effective mixed inoculum for fermentative hydrogen production from OFMSW. Mechanism of fermentative hydrogen production indicates that, among the OFMSW, carbohydrates is the most optimal substrate for fermentative hydrogen production compared with proteins, lipids and lignocelluloses. This conclusion was also substantiated by experimental results of this study. The hydrogen production potentials of rice, potato and lettuce were 134 mL/g-VS, 106 mL/g-VS, and 50 mL/g-VS respectively. The hydrogen percentages of the total gas produced from rice, potato and lettuce were 57-70%, 41-55% and 37-67%. 2008 International Association for Hydrogen Energy.

Evaluation of different culture conditions for high-level soluble expression of human cyclin A(2) with pET vector in BL21 (DE3) and spectroscopic characterization of its inclusion body structure

In this paper, we evaluated various parameters of culture condition affecting high-level soluble expression of human cyclin A, in Escherichia coli BL21(DE3), and demonstrated that the highest protein yield was obtained using TB(no glycerol) + 0.5% glucose medium at 25 degrees C. By single immobilized metal ion affinity chromatography, we got highly purified human cyclin A(2) with a yield ranged from 20 to 30 mg/L. By amyloid-diagnostic dye ThT binding and Fourier transform infrared spectroscopy, we observed a significant decrease in alpha-helix content and an increase in beta-sheet structure in cyclin A(2) inclusion body in comparison to its native protein, and confirmed the resemblance of the internal organization of cyclin A(2) inclusion body and amyloid fibrils.

Cultivation of the brown alga Sargassum horneri: sexual reproduction and seedling production in tank culture under reduced solar irradiance in ambient temperature

As a large conspicuous intertidal brown alga, individuals of Sargassum horneri can reach a length of more than 7 m with a fresh weight of 3 kg along the coasts of the Eastern China Sea. The biomass of this alga as a vital component in coastal water ecology has been well documented. In recent years, a steady disappearance of the algal biomass along the once densely populated coastal areas of the Eastern China Sea has drawn attention in China. Efforts have been made to reconstruct the subtidal algal flora or even to grow the alga by use of long-lines. As part of the efforts to establish an efficient technique for producing seedlings of S. horneri, in this investigation a series of culture experiments were carried out in indoor raceway and rectangular tanks under reduced solar irradiance at ambient temperature in 2007-2008. The investigation demonstrated that: (1) sexual reproduction of S. horneri could be accelerated in elevated temperature and light climates, at least 3 months earlier than in the wild; (2) eggs of S. horneri had the potential to be fertilized up to 48 h, much longer than that of known related species; (3) suspension and fixed culture methods were both effective in growing the seedlings to the long-line cultivation stage; and (4) the life cycle of S. horneri in culture could be shortened to 4.5 months, thus establishing this alga as an appropriate model for investigating sexual reproduction in dieocious species of this genus.

Suspension culture of gametophytes of transgenic kelp in a photobioreactor

Transgenic Laminaria japonica gametophytes producing a recombinant tissue-type plasminogen activator (rtPA) protein, which is an effective third-generation thrombolytic agent for acute myocardial infarction (AMI), were cultured in an illuminated bubble column bioreactor. A maximum final dry cell weight of 1120 mg l(-1) was obtained in batch culture with an initial dry cell weight of 126 mg l(-1) and with aeration rate of 1.2 l air min(-1) l(-1) culture, nitrate at 1.5 mM and phosphate at 0.17 mM. The yield of rtPA was 56 mu g g(-1) dry cell wt. This is the first report regarding cultivation of a transgenic macroalga in a bioreactor.

Cultivation of the brown alga Hizikia fusiformis (Harvey) Okamura: enhanced seedling production in tumbled culture

The reuse of holdfasts for regeneration of young seedlings or using wild juvenile plants as the seedling source has played the major role in commercial cultivation of the brown alga Hizikia fusiformis in East Asia over the past 20 years. The possibility of employing zygote-derived germlings for producing seedlings has been discussed in the literature, but has not yet become a reality. Three main obstacles have limited the use of zygotes as a main source of seedlings, (1) the dioecious nature of the algal life cycle which may lead to asynchronous male and female receptacle development and thus different timing of egg and spermatozoa expulsion, (2) the low attachment rate when using zygote-derived germlings with developed rhizoids from wild parental plants for seeding production, and (3) the problem of culturing young germlings in regions where water temperature is high in summer. In this investigation, shifting the timing of receptacle formation earlier than in nature was performed by tumbling the algae in a long-day tank (16-h light per day). Synchronization of egg and spermatozoa expulsion and thereafter fertilization were conducted in indoor tanks. Receptacle formation in constant long days could be shifted by 20 days earlier than in plants cultured on long lines in the open sea, or I month earlier than in plants growing on intertidal rocks. Synchronized expulsion of eggs and spermatozoon led to a high rate of fertilization. This was achieved by tumbling the male and female receptacle-bearing branchlets in the same tank at low density in high irradiance. In two independent trials, a total of 1,400,000 zygote-derived germlings were obtained from 620 g (fresh weight) female sporophytes. The germlings shed from the receptacles were at an identical developmental stage indicating high synchronization of expulsion of eggs and spermatozoon followed by fertilization. Approximately 63% ( +/-9.6%) of the germlings were shed from the receptacle between 16 and 24 It after fertilization and 20% ( +/-11.9%) remained on the receptacle for 3 days after fertilization. Germlings were seeded on string collectors before rhizoids started to elongate and the attachment efficiency was enhanced. Young seedlings reached 800 ( +/-50) mum in length in 25 days at 25 degreesC before they were transferred to open sea cultivation. These results provide the basis of a practical way of seedling production by use of zygote-derived germlings in the commercial cultivation of Hizikia fusiformis. (C) 2004 Elsevier B.V All rights reserved.