Dismutation of hydroxylamine to ammonia and nitrite by a novel, membrane-bound enzyme from the cyanobacterium Phormidium uncinatum.

Extracts from ammonium-grown phototrophic cultures of Phormidium uncinatum exhibited a catalytic and stoichiometric formation of nitrite and ammonia from hydrogen peroxide and hydroxylamine. This reaction was due to a novel enzyme (referred to as hydroxylamine dismutase), which was bound to the heavier thylakoid membranes. The enzyme was solubilized from the membranes by detergent treatment and further purified by ion-exchange chromatography. On the basis of inhibitor studies, the involvement of metal ions and sulfhydryl groups in the characteristic reaction is suggested.

Nutrients and pH regulate algicide accumulation in cultures of the cyanobacterium Oscillatoria laetevirens.

• The change in algicide synthesis in response to nutrients and pH in axenic cultures of the planktonic cyanobacterium, Oscillatoria laetevirens is reported. • Analytical and bioassay techniques were used to determine the effects of nitrogen (N), sulpher (S), phosphorus (P), magnesium (Mg2+ ), calcium (Ca2+ ) and hydrogen (H+ ) on Oscillatoria laetevirens growth, secondary metabolite accumulation and minimum inhibitory concentration in culture filtrates. • Growth and secondary metabolite production were affected by nutrient addition. High algicide production, which was not dependent on optimal growth, correlated with deficiences of magnesium and phosphate, and high pH. Algicidal activity in the culture medium was not detected following organic nitrogen addition while optimal growth was sustained; however, ammonia reduced biomass yield but did not affect algicide formation. The differential algicide concentration inside the cells and in the culture medium indicated secretion by an efflux mechanism rather than leakage from aging cells. • Algicide production by Oscillatoria laetevirens was negatively regulated by both phosphate and magnesium, and by pH. The interactive influence of nutrients and allelopathy is discussed in species succession and dominance in the maintenance of freshwater blooms.

REGULATION BY AZIDE OF HETEROCYST AND NITROGENASE IN AZIDE-RESISTANT MUTANTS OF THE CYANOBACTERIUM, NOSTOC MUSCORUM.

A novel class of azide-resistant mutants of N, muscorum is described in which azide caused inhibition of heterocyst differentiation and nitrogen fixation without causing inhibition of growth. The results indicate the utilization of azide, as a fixed nitrogen source, by the mutant strain. An increase in the ability to take up azide and in the phycocyanin/chlorophyll ratio following growth of the mutant in azide-containing medium are additional findings which support the conclusion that the mutant utilizes azide as a source of nitrogen. In the parental strain, Ca2+ -dependent and Mg2+ -dependent ATPases, and cellular nitrate reductase were inhibited by azide. The corresponding ATPases from the mutant strain were not inhibited by azide. There was evidence, in cell-free extracts, for an enzyme system which utilized azide as an electron acceptor and NADPH-ferredoxin as electron donor. The activity of this system was significantly higher (on a protein basis) in cells of the mutant grown on azide than in cells of either the parent or the mutant when grown on nitrate. It is suggested that the azide resistance of this class of mutant is due to a mutation which leads to azide resistant Ca2+ - and Mg2+ - ATPases. Such a mutation may allow an azide utilizing system, inherently present in both parent and mutant strains, to be expressed.

EVIDENCE FOR INTERGENERIC TRANSFORMATION IN FILAMENTOUS, DIAZOTROPHIC CYANOBACTERIA.

A clonal population of a mutant Nostoc muscorum, with the ability to assimilate azide as a fixed nitrogen source, was used as a source of donor DNA for transforming Anabaena doliolum from an azide-sensitive to an azide-assimilating phenotype. The donor DNA. transformed A. doliolum efficiently and the transformation process was sensitive to DNAase, resistant to RNAase A, dependent on photosynthetic light and required a specific contact period between donor DNA and recipient cells. The gene(s) responsible for azide assimilation appear to be located on chromosomal DNA and not on plasmid DNA. The transformants of A. doliolum resemble the azide assimilating mutant strain of N. muscorum in respect of (1) azide repression of heterocyst and nitrogenase formation, (2) azide-dependent increase in phycocyanin to chlorophyll ratio. These results suggest that the chromosomal DNA of mutant N. muscorum, and not its plasmid DNA, is the carrier of genetic information for its azide assimilatory phenotype and that this can be transferred efficiently to A. doliolum.

Action of an algicide from a cyanobacterium, Oscillatoria laetevirens, on photosystem II.

Oscillatoria laetevirens produces an algicide, named oscillatorin (OS), which inhibits growth of higher plants. Effect of purified oscillatorin and some 'urea-triazine type' herbicides was studied on photosystem II activity and composition of pigment protein complex in spinach thylakoid membrane. For oscillatorin the I50 at 10 microg chlorophyll concentration, inhibitor constant (Ki), specific binding sites and Hill coefficient were calculated to be 1.45, 0.15, 2.3 and 0.2 microM respectively. Metribuzin and oscillatorin affected towards the donor side and brought about identical changes in polypeptide composition of PSII complex. Further, metribuzin and atrazine exerted antagonistic and synergistic responses on oscillatorin action. Some of these parameters were also studied on weed plants to assess upon the weedicidal potential of oscillatorin.

Detoxification of a local Microcystis bloom by an algicidal antibiotic from Oscillatoria late-virens.

Mouse toxicity due to microcystin (hepatotoxin) was detected in water bloom of a local lake, comprising mainly Microcystis aeruginosa. Both toxicity and biomass content of the phytoplankton varied seasonally. Incubations with an algicidal metabolite from Oscillatoria late-virens and a herbicide 3-(3,4-dichlorophenyl)-1, 1-dimethyl urea (DCMU) caused depigmentation and detoxification in bloom samples. Algicide-incubation also encouraged growth of protozoa and bacteria, pathogenic form included.

Assessing cold chain status in a metro city of India: an intervention study.

BACKGROUND: Cold chain maintenance is an essential activity to maintain the potency of vaccines and to prevent adverse events following immunization. One baseline study highlighted the unsatisfactory cold chain status in city of Kolkata in India. OBJECTIVES: To assess the changes which occurred in the cold chain status after the intervention undertaken to improve the status and also to assess the awareness of the cold chain handlers regarding cold chain maintenance. METHODS: Intervention consisted of reorganization of cold chain points and training of health manpower in Kolkata Municipal area regarding immunization and cold chain following the guidelines as laid by Govt of India. Reevaluation of cold chain status was done at 20 institutions selected by stratified systematic random sampling after the intervention. The results were compared with baseline survey. RESULTS: Significant improvement had been observed in correct placing of cold chain equipment, maintenance of stock security, orderly placing of ice packs, diluents and vaccines inside the equipment, temperature recording and maintenance. But awareness and skill of cold chain handlers regarding basics of cold chain maintenance was not satisfactory. CONCLUSIONS: The success of intervention included significant improvement of cold chain status including creation of a designated cold chain handler. The gaps lay in non-availability of non-electrical cold chain equipment and separate cold chain room, policy makers should stress. Cold chain handlers need reorientation training regarding heat & cold sensitive vaccines, preventive maintenance and correct contingency plan.

De-regulated assimilation and over-production of amino acids in analogue-resistant mutants of a cyanobacterium, Phormidium uncinatum.

Mutant strains of Phormidium uncinatum resistant to fluoro-phenylalanine, aztryptophan, fluorotyrosine and azaleucine accumulated a wide range of amino acids, notably glutamic acid, lysine, tyrosine and phenylalanine, and exhibited de-regulated valine and phenylalanine transport. While acetohydroxy acid synthase in azaleucine-resistant mutants lost valine- and leucine-sensitivity, 3-deoxy-DXXX-arabinoheplulosonate-7-phosphate (DAHP) synthase and prephenate dehydratase in aromatic analogue-resistant strains became phenylalanine-insensitive and shikimate and prephenate dehydrogenases were activated by tyrosine. In addition, activities of nitrate-assimilating enzymes were higher in the mutants, which also exhibited increased nitrogen, protein and phycocyanin contents. The proteins in the mutants were better digested upon enzymatic-treatments and feeding trials than those of the wild type, indicating that they are usable as single-cell protein.

Evidence for ammonia as an inhibitor of heterocyst and nitrogenase formation in the cyanobacterium Anabaena cycadeae.

Growth and regulation of heterocyst and nitrogenase by fixed nitrogen sources were studied comparatively in parent and glutamine auxotrophic mutant of Anabaena cycadeae. The parent strain grew well on N2, NH+4 or glutamine while the mutant strain grew on glutamine but not on N2 or NH+4. The total lack of active glutamine synthetase in the mutant strain thus appears to be the reason for its observed lack of growth in N2 or NH+4, which explains why it is a glutamine auxotroph and at the same time shows glutamine synthetase to be the sole primary ammonia assimilating enzyme. NH+4 repression of heterocyst and nitrogenase in the mutant and the parental strains and their derepression by L-methionine-DL-sulfoximine suggest that NH+4 per se and not glutamine synthetase mediated pathway of ammonia assimilation is the initial repressor signal of heterocyst and nitrogenase in A. cycadeae.