The toxic metal ions Cu2+, Cd2+, and Pb2+ inhibited growth of the green alga Ankistrodesmus braunii and the blue-green alga (Cyanobacterium) Anabaena, strain 7120. Some concentrations of Cu lysed Anabaena 7120 at early, but not late, stages of growth. The other metals inhibited growth without causing lysis. Adding the chelating agent nitrilotriacetic acid (NTA) did not reduce, and in some cases increased, metal toxicity to Anabaena 7120. This suggests that these metals do not act on this alga only in the ionic form. When Anabaena 7120 grew in a sublethal concentrations of Cu(NO3)2 (10(-5)M) most of the Cu was found outside the cell, in nonionic form. About half the Cd was found to be cell associated when these algae grew in the presence of 10(-5)M Cd(NO3)2. Ankistrodesmus braunii bound substantial amounts of both Cd and Cu when growing in their presence. At certain Cu levels, the amount bound per cell remained virtually constant during growth. In Cd, the amount bound per cell was highest at the beginning of growth and then fell. Cell-associated metals were found in both wall plus membrane and cytoplasmic fractions of A. braunii after mechanical lysis. When these algae grew over dialysis sacs containing sediment loaded with Cd or Cu, substantial amounts of these metals left the sediment and entered the algal cultures. They were found both cell associated and in the culture medium of A. braunii. In cultures of Anabaena 7120, Cd removed from the sediment was found in both cells and culture medium, but Cu was found almost entirely in the culture medium. The effects that bloom of such algae might have on the mobilization of these metals from sediments in natural waters are discussed.

The fermentation of cellulose by monocultures of Acetivibrio cellulolyticus and cocultures of A. cellulolyticus-Methanosarcina barkeri, A. cellulolyticus-Desulfovibrio sp., and A. cellulolyticus-M. barkeri-Desulfovibrio sp. was studied. The monoculture produced ethanol, acetate, H2, and C02. More acetate and less ethanol was formed by the cocultures than by the monoculture. Acetate was utilized by M. barkeri in coculture with A. cellulolyticus after a lag period, whereas ethanol was metabolized by the sulfate reducer only under conditions of low H2 partial pressure, i.e., when cocultured with A. celluloyticus-M. barkeri or when grown together with the methanogen. Only the three-component culture carried out the rapid conversion of cellulose to C02 and methane. Furthermore, this culture hydrolyzed the most cellulose-85% of that initially present. This amount was increased to 90% by increasing the population of M. barkeri in the triculture. Methane production was also increased, and a quicker fermentation rate was achieved.

In the xylose fermentation of Bacillus polymyxa strain 9035, best 2,3-butanediol yields were obtained with 1.0% yeast extract, 4-6% xylose, shaking at 125 rpm and incubation at 30°C.Under these conditions, mannose, galactose, L-arabinose, cellobiose, starch and glucose were readily metabolized and yielded significant amounts of diol.Diol production from xylan was also demonstrated. In addition, the screening of a number of B___tpolymyxa strains on xylose revealed that only strains 9031-1 and 9035 used xyiose extensively and produced significant amounts of diol.The latter strain proved best under scaled-up conditions.

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The effects of brief exposure to, or growth in the presence of, lethal and sublethal concentrations of Cu(NO)2 and Cd(NO3) on the ultrastructure of the blue-green algaAnabaena 7120 and the green algaAnkistrodesmus braunii were studied. Exposure to increasing amount of both metal ions led to the appearance of larger proportions of electron-dense cells whose organelles were less well defined than those of untreated cells. Metal-treated cells ofAnabaena 7120 became distorted. Some had a corrugated appearance. Others lysed, leaving a much larger proportion of heterocysts. Such heterocysts were often empty or had a curious collapsed appearance. Growth ofA. braunii in the presence of 10(-4) M Cu(NO2)2 produced substantial numbers of multinucleate giant cells with thick walls; such cells result from repeated mitotic division without subsequent cytokinesis. The giant cells contained centrioles, structures not as yet found in normal cells of the genusAnkistrodesmus. Some nuclei of giant, but not of normal, cells contained deep indentations that appeared as "holes" in cross section. Some giant cells also contained triple parallel strands of endoplasmic reticulum which extended across much of the cell, connecting to the nuclear envelope. Some ultrastructural changes were also noted in algal cells grown over sediment containing Cu or Cd, but these were generally less severe than those occurring when metal ions were added directly to the algal cultures.