Toxicity of parathion-methyl to cells, akinetes and heterocysts of the cyanobacterium Cylindrospermum, sp. and the probit analysis of toxicity

The toxicity of a commercial formulation of the insecticide parathion‐methyl to the N2‐fixing filamentous cyanobacterium (blue‐green alga) Cylindrospermum, sp. was studied. A concentration of parathion‐methyl of 0.5 ppm caused growth increase in liquid growth media. The minimum inhibitory concentration of parathion‐methyl for both types (N₂, fixing and nitrate supplemented) of liquid and solid media was 1.0 ppm. LC₅₀ values were: 4.4 ppm (liquid, N₂, fixing), 5.5 ppm (liquid, nitrate supplemented), 3.3 ppm (agar, N₂‐fixing) and 4.0 ppm (agar, nitrate supplemented). LC100 values for N₂‐fixing liquid and both types of agar media were 10.0 ppm, while for the liquid nitrate supplemented medium the LC₁₀₀ was 12.0 ppm. Both akinete (spore) formation and germination were inhibited below the highest permissive concentration of 8.0 ppm, with the insecticide incorporated in the agar media. In soil, the LC50 and LC100 values for parathion‐methyl were 13.6 and 30 ppm, respectively. Both the dehydrogenase activity of heterocysts (monitored by 2,3,5‐triphenyl tetrazolium chloride reduction) and the nitrogen concentration of cultures (estimated by the micro‐Kjeldahl method) were affected by the insecticide, but the latter (N₂‐fixation) was more sensitive. The Kruskal‐Wallis H test on the numbers of vegetative cells in the filaments revealed that the insecticide significantly affected the division of vegetative cells. The cyanobacterium could detoxify the growth medium containing high levels (30 and 40 ppm) of the insecticide in short‐term exposures at the expense of cell viability.     

Notes on the growth and sporulation of a natural population of Aphanizomenon flos-aquae

Aphanizomenon flos-aquae contributed regularly to the summer phytoplankton in Kinnego Bay, a highly eutrophic part of Lough Neagh. Growth of the alga was variable in different years, and was apparently related to the duration of depletion of dissolved nitrate-nitrogen in the water. Although four growth pulses occurred during the study period, sporulation was only observed during one of these, when the population was very dense although evenly distributed down the water column. It seems that Aphanizomenon overwintered in Kinnego Bay as vegetative filaments and that production of akinetes was not necessary for perennation of the species. The observed sporulation in 1973 may have been induced by the high pH values caused by dense phytoplankton crops; no other measured environmental factors were particularly unfavourable to growth of Aphanizomenon at this time.     

AKINETE GERMINATION IN ANABAENA CIRCINALIS (CYANOPHTA)

Akinetes of a clonal culture of Anabaena circinalis Rabenhorst from Mt. Bold reservoir (eutrophic), South Australia, were isolated and the effects of light, phosphorus, and nitrogen availability on their germination were investigated. Light was required but there was no significant difference in percentage of germination after 72 h if akinetes were incubated in ASM-1 medium at irradiances of 15, 30, or 50 μmol.m^-2.s^-1. Maximum akinete germination occurred by 48 h. Nitrogen was not required, as 88% of akinetes germinated in the flasks without combined nitrogen added to the medium and without N₂ in the air. NH₄⁺-N at 28 mg N.L^-1 completely suppressed germination, whereas 28 mg NO₃ N.L^-1 had no effect relative to the controls without nitrogen. Phosphorus was required, and at 48 h percentage of germination in the flasks with 0.6 mg P.L^-1 added (78%) was significantly greater than in the flasks with 0.06 P.L^-1 (58%) and 0 mgP.L^-1 (24%) added. Germlings in the 0 mg P.L^-1 flasks were only 2–4 cells long and stunted in appearance, whereas germlings at all other P concentrations were 8–16 cells long. It is likely that the isolation process exposed some akinetes to intracellular phosphorus released from lysing vegetative cells, but this was insufficient to allow normal development in the 0 mg P.L^-1 flasks. The plot of percentage of germination vs. initial phosphorus concentration, in the medium showed a relationship analogous to Michaelis-Menten nutrient uptake kinetics, suggesting that a specific membrane-bound enzyme system(s) is involved, with phosphorus as the substrate. The half saturation value (K_S) for germination was 50 μg P.L^-1.     

POLYSACCHARIDES FROM THE ENVELOPES OF HETEROCYSTS AND SPORES OF THE BLUE-GREEN ALGAE ANABAENA VARIABILIS AND CYLINDROSPERMUM LICHENIFORME1

The polysaccharides from the envelopes of heterocysts of Cylindrospermum licheniforme Kütz., and of heterocysts and spores of Anabaena variabilis Kütz., like those from the differentiated cells of Anabaena cylindrica Lemm., have a 1,3-linked backbone consisting of glucosyl and mannosyl residues in a molar ratio of approximately 3:1. As is the case with A. cylindrica the polysaccharides from A. variabilis and from the heterocysts of C. licheniforme have terminal xylosyl and galactosyl residues as side branches. In addition, the polysaccharide from C. licheniforme resembles that from A. cylindrica in having terminal mannosyl residues as side branches (absent from A. variabilis). The polysaccharides from A. variabilis resemble that from A. cylindrica in having glucose-containing side branches (absent from the heterocyst polysaccharide from C. licheniforme), but in contrast to the polysaccharides from the other two species they also have terminal arabinosyl residues as side branches. All of the polysaccharides mentioned appear to be structurally related; we present tentative structures for those not previously investigated. In contrast, the envelope of spores of C. licheniforme contains only a largely 4-linked galactan. The bulk of this envelope is not polysaccharide in nature, and contains aromatic groups.     

AKINETE PRODUCTION OF ANABAENA LEMMERMANNII AND A. CYLINDRICA (CYANOPHYCEAE) IN NATURAL POPULATIONS OF N‐ AND P‐LIMITED COASTAL MESOCOSMS1

A strong biomass increase of two Anabaena species was observed in natural plankton community enclosed into nine large mesocosms (51 m³) and manipulated with mineral nutrients and an organic carbon source during a 3‐week period in the coastal Baltic Sea. The water column and settled material from the bottom of the mesocosms were sampled at 2‐day intervals. Planktonic populations of Anabaena lemmermannii Richter and A. cylindrica Lemmermann and sedimentation rates of akinetes to the bottom were quantified. Comparing mesocosms with artificially induced nitrogen and phosphorus limitation, we found that during the third week of the experiment, the population size of A. lemmermannii was clearly higher in nitrogen‐limited units (by a factor of 2.4), whereas the production rate of akinetes was higher in the phosphorus‐limited units (by a factor of 2.5). Input of freshly produced A. lemmermannii akinetes to the benthos was on average 15 × 10⁶ and 6 × 10⁶ cells· m^?2·d^?1 in the P^? and N^? limited mesocosms, respectively. Our estimates of specific akinete production rate of A. lemmermannii in P^? and N^? limited mesocosms revealed an even larger divergence (a factor of 5.5), being on average 2.4 and 0.4 akinetes·10^?3 vegetative cells^?1·d^?1, respectively. The phosphorus addition effectively reduced akinete production of A. lemmermannii. Differences in the nutrient manipulation had no apparent effect on the biomass and akinete production of A. cylindrica. The akinete production pattern of A. cylindrica revealed a 1‐week delay compared with the vegetative population peak, whereas such a delay was not obvious in A. lemmermannii.