The Oxidation of Iso-citrate by a Particulate Fraction Obtained from Swede Storage Tissue

A particulate fraction has been prepared from swede storagetissue which possesses both an NAD and an NADP linked iso-citricdehydrogenase. Such preparations have been used to study thepathway of oxidation of iso-citrate and the reduced coenzyme—cytochromec. reductase activity. The activity of the enzyme systems associatedwith the particles varies with pH, at pH 7.1 the NAD linkediso-citric dehydrogenase exhibits optimal activity whilst theoptimum pH for the NADP linked dehydrogenase is pH 8.3. At eitherpH the major pathway of oxidation of iso-citrate appears tobe through the NAD linked iso-citric dehydrogenase, NADH₂-cytochromec. reductase system. The preparations are apparently unableto transfer electrons from reduced coenzyme, produced by dehydrogenaseactivity, to cytochrome c. at the same rate as they are ableto transfer electrons from reduced coenzyme supplied as a substrate.The results are discussed in relation to mitochondrial structureand a comparison is made with the results of work carried outon the iso-citric dehydrogenases of animal tissues.     

Identification and Characterization of Bacteria in a Selenium-Contaminated Hypersaline Evaporation Pond

Solar evaporation ponds are commonly used to reduce the volume of seleniferous agricultural drainage water in the San Joaquin Valley, Calif. These hypersaline ponds pose an environmental health hazard because they are heavily contaminated with selenium (Se), mainly in the form of selenate. Se in the ponds may be removed by microbial Se volatilization, a bioremediation process whereby toxic, bioavailable selenate is converted to relatively nontoxic dimethylselenide gas. In order to identify microbes that may be used for Se bioremediation, a 16S ribosomal DNA phylogenetic analysis of an aerobic hypersaline pond in the San Joaquin Valley showed that a previously unaffiliated group of uncultured bacteria (belonging to the order Cytophagales) was dominant, followed by a group of cultured γ-Proteobacteria which was closely related to Halomonas species. Se K-edge X-ray absorption spectroscopy of selenate-treated bacterial isolates showed that they accumulated a mixture of predominantly selenate and a selenomethionine-like species, consistent with the idea that selenate was assimilated via the S assimilation pathway. One of these bacterial isolates (Halomonas-like strain MPD-51) was the best candidate for the bioremediation of hypersaline evaporation ponds contaminated with high Se concentrations because it tolerated 2 M selenate and 32.5% NaCl, grew rapidly in media containing selenate, and accumulated and volatilized Se at high rates (1.65 μg of Se g of protein⁻¹ h⁻¹), compared to other cultured bacterial isolates.     

Enzymatic Oxidation of Tetramethyl-p-Phenylenediamine and p-Phenylenediamine by the Electron Transport Particulate Fraction of Azotobacter vinelandii

The ability of the electron transport particulate fraction of Azotobacter vinelandii strain O to oxidize tetramethyl-p-phenylenediamine (TMPD) and p-phenylenediamine (PPD) was examined in detail. The highest specific activity for TMPD and PPD oxidation concentrated in the A. vinelandii O R(3) fraction. The A. vinelandii O R(3) fraction was used to develop a standard manometric assay which gave optimal oxidation rates for both of these dyes. The conditions of the assay and all essential related enzymatic kinetic parameters are presented. Other para derivatives of phenylenediamines also were oxidized readily, whereas ortho and meta derivatives were not. Hydroquinone, p-hydroxybenzoic acid, p-cresol, tyrosine, pyrogallol, pyrocatechol, and diphenylamine were not able to serve as electron donors for the A. vinelandii O R(3) system. The probable involvement of a particle-bound cytochrome oxidase is indicated by the marked sensitivity of both TMPD and PPD oxidation to cyanide, axide, phenylhydrazine, hydroxylamine, and, to a lesser degree, carbon monoxide.     

Decomposition of Organic Mercurial Compounds by Mercury-resistant Bacteria

Isopentyl isovalerate and anisic acid were first isolated and identified from the Japanese peppermint oil. The former compound possesses characteristic apple-like oder. α-Bourbonene, menthofurolactone, and β-caryophyllene epoxide were also isolated and identified from the oil of Shubi, a newly registered Japanese peppermint.     

Carboxymethyl Cellulose Decomposition by Intestinal Bacteria of Cockroaches

The effect of freeze-drying on phenotypic reversion of amino acid auxotrophy to prototrophy was studied in Escherichia coli. In a radioresistant strain, E. coli H/r 30 (uvr+ exr+), which can repair the deoxyribonucleic acid damaged due to freeze-drying, an increased mutation frequency from auxotrophy to prototrophy was observed with increased time of freeze-drying of the cells. On the other hand, in a radiosensitive strain, E. coli NG 30 (recA), which cannot repair the damaged deoxyribonucleic acid due to a lack of repair enzyme system, no significant reversion occurred, although the survival rate was very low. The rate of phenotypic reversion dut to freeze-drying in both E. coli RIMD 0509109 (uvr+ exr+) and RIMD 0509115 (uvr exr+) was almost the same, indicating that the phenomenon is independent of the uvr character. From these results it is concluded that mutation was induced in E. coli cells during the rehydration when the damaged deoxyribonucleic acid was repaired by exr character of the cells. Thus, we propose that a serious consideration should be paid to the freeze-drying technique to preserve bacterial cells.     

Numerical Taxonomy of Coryneform Bacteria Isolated from Pond-Reared Shrimp (Penaeus aztecus) and Pond Water

Gram-positive, catalase-positive, nonsporeforming, pleomorphic rods isolated from pond-reared shrimp and pond water were compared with type cultures of the Corynebacteriaceae. Classification of the type cultures based on 66 cell and colony characters proved comparable to one based on 163 morphological, biochemical, and physiological characters. This similarity was not observed with pond isolates. With the aid of numerical analysis, pond isolates could be placed into six major groups based on certain biochemical and physiological tests. Coryneform bacteria isolated from shrimp and water exhibited little similarity to the type cultures. The pond isolates probably are members of the Corynebacteriaceae not previously studied in detail.     

Production of Urea by Bacterial Decomposition of Organic Matter Including Phytoplankton

A concentrated colony of Fragilaria crotonensis collected from the surface water of Lake Suwa, which is one of the typical eutrophic lakes in Japan, and organic matter contained in untreated surface water from the same lake were subjected to aerobic decomposition by bacteria in a dark room at a temperature of 20 ± 3 °C. An exponential increase of urea with time was recorded in both of the experiments. The apparent rate constants of urea production were calculated to be 0.083 day⁻¹ for decomposition of F. crotonensis and 0.051 day⁻¹ for decomposition of the organic matter contained in the untreated surface water. This study suggests that urea production by bacterial decomposition of organic matter, including phytoplankton, may be an important source of urea in natural waters under certain conditions.     

Total Counts of Marine Bacteria Include a Large Fraction of Non-Nucleoid-Containing Bacteria (Ghosts)

Counts of heterotrophic bacteria in marine waters are usually in the order of 5 x 10(sup5) to 3 x 10(sup6) bacteria ml(sup-1). These numbers are derived from unspecific fluorescent staining techniques (J. E. Hobbie, R. J. Daley, and S. Jasper, Appl. Environ. Microbiol. 33:1225-1228, 1977; K. G. Porter and Y. S. Feig, Limnol. Oceanogr. 25:943-948, 1980) and are subsequently defined as total counts of bacteria. In samples from the Baltic Sea, the North Sea (Skagerrak), and the northeastern Mediterranean Sea, we found that only a minor fraction (2 to 32%) of total counts can be scored as bacteria with nucleoids. Lack of DNA no doubt means inactive cells; therefore, a much lower number of bacteria that grow at rates higher than those previously estimated must be responsible for the measured bacterial production in these seas. The remaining bacterium-sized and/or -shaped particles included in total counts may be cell residues of virus-lysed bacteria (ghosts) or remains of protozoan grazing.     

Distribution and Solubilization of Particulate Gluconate Dehydrogenase and Particulate 2-Ketogluconate Dehydrogenase in Acetic Acid Bacteria

The distribution of two particulate enzymes, gluconate dehydrogenase (GDH) and 2-ketogluconate dehydrogenase (2KGDH), was investigated with cell free extract through 26 strains of genus Acetobacter and genus Gluconobacter. GDH activity was found in the cell free extracts from all strains of genus Gluconobacter and two species of genus Acetobacter, A. aceti and A. aurantium. High activity of 2KGDH was also found in the pigment-producing strains of genus Gluconobacter.

Best solubilization of particulate enzymes was attained with the highest recovery when 10 mg of Triton X–100 and 30 mg of protein of particulate fractions in 1 ml of 0.01 m phosphate buffer, pH 6.0, are incubated for 9 hr at 5°C with continuous stirring.

By comparison of the total enzyme activity of particulate enzymes with that of NAD(P)-linked enzymes in the cell free extract, it was obvious that the formation of ketogluconates by particulate enzymes was much more predominant, roughly over 100 times higher, as that of NAD(P)-linked enzymes.     

Design and Application of a Solar Mobile Pond Aquaculture Water Quality-Regulation Machine Based in Bream Pond Aquaculture

Bream pond aquaculture plays a very important role in China’s aquaculture industry and is the main source of aquatic products. To regulate and control pond water quality and sediment, a movable solar pond aquaculture water quality regulation machine (SMWM) was designed and used. This machine is solar-powered and moves on water, and its primary components are a solar power supply device, a sediment lifting device, a mechanism for walking on the water’s surface and a control system. The solar power supply device provides power for the machine, and the water walking mechanism drives the machine’s motion on the water. The sediment lifting device orbits the main section of the machine and affects a large area of the pond. Tests of the machine’s mechanical properties revealed that the minimum illumination necessary for the SMWM to function is 13,000 Lx and that its stable speed on the water is 0.02–0.03 m/s. For an illumination of 13,000–52,500 Lx, the sediment lifting device runs at 0.13–0.35 m/s, and its water delivery capacity is 110–208 m³/h. The sediment lifting device is able to fold away, and the angle of the suction chamber can be adjusted, making the machine work well in ponds at different water depths from 0.5 m to 2 m. The optimal distance from the sediment lifting device to the bottom of the pond is 10–15 cm. In addition, adjusting the length of the connecting rod and the direction of the traction rope allows the SMWM to work in a pond water area greater than 80%. The analysis of water quality in Wuchang bream (Parabramis pekinensis) and silver carp (Hypophthalmichthys molitrix) culture ponds using the SMWM resulted in decreased NH₃⁺–N and available phosphorus concentrations and increased TP concentrations. The TN content and the amount of available phosphorus in the sediment were reduced. In addition, the fish production showed that the SMWM enhanced the yields of Wuchang bream and silver carp by more than 30% and 24%, respectively. These results indicate that the SMWM may be suitable for Wuchang bream pond aquaculture in China and that it can be used in pond aquaculture for regulating and controlling water quality.     

Spatiotemporal Distribution of Marine Magnetotactic Bacteria in a Seasonally Stratified Coastal Salt Pond

The occurrence and distribution of magnetotactic bacteria (MB) were studied as a function of the physical and chemical conditions in meromictic Salt Pond, Falmouth, Mass., throughout summer 2002. Three dominant MB morphotypes were observed to occur within the chemocline. Small microaerophilic magnetite-producing cocci were present at the top of the chemocline, while a greigite-producing packet-forming bacterium occurred at the base of the chemocline. The distributions of these groups displayed sharp changes in abundance over small length scales within the water column as well as strong seasonal fluctuations in population abundance. We identified a novel, greigite-producing rod in the sulfidic hypolimnion that was present in relatively constant abundance over the course of the season. This rod is the first MB that appears to belong to the γ-Proteobacteria, which may suggest an iron- rather than sulfur-based respiratory metabolism. Its distribution and phylogenetic identity suggest that an alternative model for the ecological and physiological role of magnetotaxis is needed for greigite-producing MB.