Mechanism of bactericidal action of phenethyl alcohol inEscherichia coli

The mechanism of bactericidal action of phenethyl alcohol (PEA) inE. coli, which was previously demonstrated to be dependent on protein synthesis, has been investigated. Mutants resistant to PEA were selected, but the resistance observed was associated with a change in permeation. PEA effects on DNA, RNA, and protein synthesis were studied with bacteriostatic and bactericidal concentrations Similar results (inhibition of DNA synthesis and decrease in RNA synthesis) were obtained with lethal concentrations of PEA in cells pretreated with chloramphenicol, and with bacteriostatic concentrations of PEA in unpretreated cells. The PEA intracellular accumulation reached a maximum within 4 min and was not inhibited by KCN or by 2,4-dinitrophenol. The presence of phenylacetaldehyde was demonstrated in both stationary and exponential growth phase cells exposed to PEA but not in cells pretreated with chloramphenicol. These results suggested that the bactericidal mechanism of action of PEA involves its conversion into the corresponding aldehyde.     

Isolation and purification of siderophores produced by cyanobacteria,Synechococcus sp. PCC 7942 andAnabaena variabilis ATCC 29413

New siderophores were isolated and purified from the spent growth medium of the cyanobacteriaSynechococcus sp. PCC 7942 (Anacystis nidulans R2) andAnabaena variabilis ATCC 29413 by solvent extraction and thin-layer chromatography. For each species the siderophore was released into the medium when the cells were grown at low iron concentrations and was not found in the medium of cells grown in iron-sufficient medium. Through a series of biological and chemical tests, combined with spectral analysis, the dihydroxamate nature of each siderophore was confirmed. The siderophores produced bySynechococcus sp. PCC 7942 andA. variabilis had distinct relative molecular masses of 310–313 Da and 520–525 Da, respectively. Neither of the two strains produced Arnow-positive extracellular organics, which indicate the excretion of extracellular catechol-type siderophores.     

Differential and kinetic effects of cell cycle inhibitors on neoplastic and primary astrocytes

Alterations in cell cycle regulation underlie the unrestricted growth of neoplastic astrocytes. Chemotherapeutic interventions of gliomas have poor prognostic outcomes due to drug resistance and drug toxicity. Here, we examined the in vitro growth kinetics of C6 glioma (C6G) cells and primary astrocytes and their responses to 2 phase-specific inhibitors, lovastatin and hydroxyurea. C6G cells demonstrated a shorter G₁ phase and an earlier peak of DNA synthesis in S phase than primary astrocytes. As C6G cells and primary astrocytes re-entered the cell cycle in the presence of lovastatin or hydroxyurea, they exhibited different sensitivities to the inhibitory effects of these agents, as measured by [³H]-thymidine incorporation. Compared to primary astrocytes, C6G cells were more sensitive to lovastatin, but less sensitive to hydroxyurea. Studies using 2 different paradigms of exposure uncovered dramatic differences in the kinetics of DNA synthesis inhibition by these 2 agents in C6G cells and primary astrocytes. One notable difference was the ability of C6G cells to more easily recover from the inhibitory effects of hydroxyurea following short exposure. Our results provide insight into C6 glioma drug resistance as well as the inhibitory effects of these 2 phase-specific inhibitors and their chemotherapeutic potential.     

Effect of iron deficiency on ferredoxin levels in Anabaena variabilis PCC 6309

Cultures of the nitrogen-fixing cyanobacterium Anabaena variabilis PCC 6309 were grown under several iron concentrations, and changes in growth rates and chlorophyll and phycocyanin concentrations were determined. The total amount of ferredoxin present in the cells was found to be dependent on the concentration of iron in the media, as was the pattern distribution of the two different forms of the iron-sulfur protein described in this organism. Flavodoxin was not found to be present in these cells even when they were grown in the absence of added iron, indicating that this flavoprotein does not replace ferredoxin in this particular strain.     

Melafen action on the growth and physiological parameters of phototrophic and heterotrophic microorganisms

Melafen, a synthetic plant growth regulator, stimulates the growth of periodic cultures of cyanobacteria Anacystis nidulans and Anabaena variabilis and green microalgae Chlorella vulgaris by 30–80%, when applied within the concentration range of 10⁻⁸–10⁻⁶ M. If the melafen concentration lies out of this range, it is either ineffective or inhibits the growth of these microorganisms. The growth and development of green microalgae Dunaliella maritima are stimulated, and the cell growth of heterotrophic bacteria Pseudomonas diminuta is inhibited in a wide range of the increased melafen concentrations. This agent also stimulates the growth of dialyzed phototrophic cultures and the formation of heterocysts and vegetative cells in the filaments of Anabaena variabilis, increases the redox potential of a cultivation medium, and reduces a carbohydrate excretion from the cells of phototrophic microorganisms. It is supposed that melafen activates the processes of photosynthesis and atmospheric nitrogen fixation.     

Sensitivity of a Bacteroides melaninogenicus strain to monosaccharides: effect on enzyme induction.

The inhibition of growth in Bacteroides melaninogenicus by sugars in described. Monosaccharides such as D-glucose, D-galactose, D-mannose, and D-fructose are inhibitory at low concentrations, whereas the disaccharides sucrose and lactose are not inhibitory even at high concentrations. The major inhibitory effect of the sugar is found during the transition of lag to logarithmic growth phases. There was no primary effect of D-glucose on protein, ribonucleic acid, or deoxyribonucleic acid synthesis on cells in transition from lag to logarithmic growth. However, the addition of glucose or galactose completely abolished the induction of 3-ketodihydrosphingosine synthetase by vitamin K in vitamin K-depleted cells. Futhermore, in cells which were not vitamin K depleted, the level of this enzyme was drastically reduced by the addition of the sugar. Cyclic adenosine 5-monophosphate was unable to reverse the growth inhibition produced by glucose. In actively growing cultures, addition of sugar slows the growth rate. In these experiments the level of 3-ketodihydrosphingosine synthetase fell only after the cells had assumed the slower rate of growth. There were two indications that D-galactose was more inhibitory than D-glucose; in the presence of 0.1% D-galactose cells in lag phase did not show the increase in turbidity found in similar cells placed in medium with 0.1% D-glucose, and also D-galactose caused a greater decrease in the growth rate of actively growing cultures than was found with D-glucose. These studies suggest that the inhibitory effect of monosaccharides in lag leads to logarithmic growth transition can be ascribed to an effect on enzyme induction. On the other hand, the ability of many monosaccharides to inhibit growth, and the greater inhibitory property of D-galactose compared with D-glucose, suggests that other mechanisms may be operative as well.     

Induction of sexual co-flocculation of heterothallic fission yeast (Schizosaccharomyces pombe) cells by mating pheromones

Heterothallic fission yeast (Schizosaccharomyces pombe) cells preincubated with sex pheromone, P- or M-factor of the obverse mating-type cells, in mannose synthetic medium (MSM) results in remarkably increased sexual co-flocculation with obverse mating-type cells almost without time lag, i.e., within 10 min. By contrast, comparable flocculation requires over 1 h if untreated control cells are mixed with obverse mating-type cells. The agglutinin of P cells is more inducible than that of M cells. These pheromonal inductions of sexual co-flocculation are inhibited by the addition of cycloheximide or tunicamycin during preincubation but not by chloramphenicol or hydroxyurea. These results demonstrate that, in addition to (a) the repression of cell division (G1 arrest) and (b) the activation of cell wall autolytic processes (mating-specific elongation of cells: formation of their conjugation tubes), mating pheromones of fission yeast have another important role; (c) to induce sexual co-flocculation (agglutinability). Using our experimental system of preincubation with sexual pheromones, we show that M-agglutinin is heat-stable and its induction is inhibited by tunicamycin, but that P-agglutinin is heat-labile and its induction is only partially inhibited by tunicamycin.     

Formation of vesiculated large bodies of Staphylococcus aureus L-form in a liquid medium

A method is described in which cells of Staphylococcus aureus can be converted to vesiculated large bodies of L-form. When coccal cells were incubated in a liquid growth medium containing D-cycloserine, N-acetylmuramidase and subtilisin, a large number of vesiculated large bodies were formed. Electron microscopy revealed that development of internal vesicles arose after 6 hr of incubation.. When growth inhibitory concentrations of rifampicin, novobiocin, or chloramphenicol were added to the culture at 6 hr of incubation, small-sized nonvesiculated bodies were produced instead of vesiculated forms. The viability of cultures was reduced by rifampicin and novobiocin but not by chloramphenicol.     

Effects of various compounds on growth of yeast cells of Histoplasma capsulatum

Summary A number of compounds were screened for their effects on growth of the yeast cells ofHistoplasma capsulatum. Included were penicillin and related compounds, sulfhydryl inhibitors, various organic sulfur compounds recently synthesized for the first time, and compounds structurally related to the required metabolites, thiamine and cystine or cysteine. Cephalothin was the only one of the penicillin related compounds which inhibited growth. This occurred only when a high concentration (8.3 × 10^–4 M) was used. Of the analogues of cystine tested, allylglycine had the greatest inhibitory effect on growth of the yeast cells in the synthetic medium, but it failed to inhibit growth in a complex medium containing peptones and plasma. Among the sulfhydryl inhibitors, the maleimides were the most effective, producing complete inhibition of growth in the peptone medium at 10µg/ml or less. At subinhibitory concentrations the cultures tended to become mycelial. The action of the maleimides was reversed by cystine over a range of concentrations. At low concentrations, some of the disulfide derivatives of thiamine stimulated growth equally as well as thiamine, but at concentrations of 100 to 150µg/ml, they completely inhibited growth. On the basis of results obtained to date, three classes of the new organic sulfur compounds being tested offer promise as sources of potentially useful chemotherapeutic agents. These classes, which differ widely in structure, are as follows: the benzyl decylaminoethyl disulfides, the acyl disulfides, and the trithiopercarbamates.This investigation was supported by Public Health Service Research Grant AI-03524 from the National Institute of Allergy and Infectious Diseases.     

Pretreatment strategies for microbial valorization of bio‐oil fractions produced by fast pyrolysis of ash‐rich lignocellulosic biomass

This work evaluates a biorefinery approach for microbial valorization of bio‐oil fractions produced by fast pyrolysis of ash‐rich lignocellulosic biomass. Different methods are presented for the pretreatment of the low‐sugar complex bio‐oil consisting of organic condensate (OC) and aqueous condensate (AC) to overcome their strong inhibitory effects and unsuitability for common analytical methods. Growth of Pseudomonas putida KT2440, which was chosen as a reference system, on untreated bio‐oil fractions was only detectable using solid medium with OC as sole carbon source. Utilization of a pretreated OC which was filtered, autoclaved, neutralized and centrifuged enabled growth in liquid medium with significant remaining optical instability. By subjecting the pretreated fractions to solid phase extraction, more stable and less inhibitory bio‐oil fractions could be obtained enabling the appliance of common analytical methods. Furthermore, this pretreatment facilitated growth of the applied reference organism Pseudomonas putida KT2440. As there is currently no convincing strategy for reliable application of bio‐oil as a sole source of carbon in industrial biotechnology, the presented work depicts a first step toward establishing bio‐oil as a future sustainable feedstock for a bio‐based economy.     

Tick saliva regulates migration, phagocytosis, and gene expression in the macrophage-like cell line, IC-21

We studied the effects of tick saliva on cell migration, cell signaling, phagocytosis, and gene expression in the murine macrophage cell line, IC-21. Saliva increased both basal- and platelet-derived growth factor (PDGF)-stimulated migration in IC-21 cells. However, saliva did not affect PDGF-stimulated extracellular signal-regulated kinase (ERK) activity. Zymosan-mediated interleukin-1 receptor associated kinase (IRAK) activity increased when cells were pretreated with saliva. Saliva suppressed phagocytosis of zymosan particles by IC-21 cells. An RT² Profiler™ PCR Array revealed that saliva regulates gene expression in a manner consistent with an immune response skewed toward a Th2 reaction, which is characterized by production of anti-inflammatory cytokines IL-4 and IL-10. Our results using IC-21 cells suggest that Dermacentor variabilis has evolved a mechanism for regulating macrophage function, which may contribute to the tick’s ability to modulate immune function.     

Tolerance to Putrescine Toxicity in Chinese Hamster Ovary Cells Is Associated with Altered Uptake and Export

When Chinese hamster ovary (CHO) cells were cultured with low concentrations of putrescine (< 5 mM) their cell cycle time increased significantly and a fraction of the cells died. A cell line tolerant to the cytotoxic and growth inhibitory effects of millimolar concentrations of putrescine was developed by growing CHO cells over many months in increasing concentrations of the polyamine. A putrescine-tolerant cell line was obtained which was capable of growing in concentrations up to 25 mMputrescine and displayed growth and cell division rates similar to the original untreated/parental CHO cells. The tolerant cells grown in putrescine displayed relatively high intracellular putrescine yet the cell-associated putrescine concentration was estimated to be 10-fold less than the culture medium level. This high concentration of cellular putrescine diminished within 60 min when the cells were changed to non-putrescine-containing media. The putrescine-tolerant phenotype was further characterized in regards to the mechanisms involved in putrescine uptake, efflux, and biosynthesis. The parental and tolerant cell lines had similar or identical levels of cellular spermidine and spermine and no differences in the acetylated polyamine pools or diamine oxidase activity. The activity of ornithine decarboxylase was also similar in the two cell lines in both the presence and the absence of ornithine. The tolerant cells, however, had a decreased uptake rate for putrescine. The tolerant cell line also showed a greatly enhanced ability to export putrescine, especially when treated with ornithine, suggesting that an upregulated polyamine export system may be present in the tolerant cells which could be responsible for the increased cell survival in high putrescine concentrations. The data are discussed in regard to the potential for identifying the transport protein(s) responsible for the maintenance of nontoxic intracellular concentrations of putrescine in a tolerant cell line grown in putrescine.     

Chloramphenicol inhibition of Pythium ultimum and Rhodotorula glutinis

Summary The growth of Rhodotorula glutinis is inhibited by both D-threo chloramphenicol and an L-threo isomer of chloramphenicol (lacking the dichloroacetyl group), causing an increase in the mean generation time, in a variety of media, approximately proportional to the concentration of antibiotic. The antibiotic is not removed from the growth medium in any quantity during this inhibition of growth. The oxygen uptakes of normal and chloramphenicol-grown cells of R. glutinis are similar when expressed on a dry weight basis. The oxygen uptake of normal and L-threo isomer-grown cells is strongly inhibited by antimycin A, whereas D-threo chloramphenicol-grown cells are unaffected. There was no evidence to suggest that any uncoupling of phosphorylation occurred with either isomer. Pythium ultimum mycelium also showed similar oxygen uptakes per unit dry weight whether grown in the presence or absence of D-threo chloramphenicol. The D-threo chloramphenicol-grown mycelium was also insensitive to antimycin A in contrast to the normal mycelium which was strongly inhibited. P. ultimum grows slowly in the presence of 100 g/ml D-threo chloramphenicol in a glucose salts medium, but is completely inhibited by a similar concentration in a glycerol salts medium. The L-threo isomer does not inhibit the growth of P. ultimum.The mitochondria of Rhodotorula glutinis show a progressive disorganization when grown in the presence of increasing concentrations of D-threo chloramphenicol up to 1000 g/ml. There is an associated over synthesis of cell wall material in the higher concentrations of the antibiotic. The L-threo isomer produces no obvious fine structural abnormalities even at concentrations of 1000 g/ml.     

Lysozyme-induced inhibition of the lymphocyte response to mitogenic lectins

Both human lysozyme (HL) and hen egg white lysozyme (HEWL) inhibited the proliferative response of peripheral blood lymphocytes to T cell mitogens such as the lectins phytohemagglutinin and concanavalin A. This inhibition was observed both when HL or HEWL was added to the lymphocyte cultures in combination with phytohemagglutinin or concanavalin A and when lymphocytes were pretreated with either lysozyme and extensively washed prior to culture with mitogens. Under both conditions, the effects were strictly dose dependent; the lysozyme concentrations yielding maximal inhibitory effect were 5 micrograms/ml for HL and 1 microgram/ml for HEWL, while both lower and higher concentrations were less effective. Specific antilysozyme rabbit sera completely prevented the inhibitory effects of both HL and HEWL on the proliferative response of lymphocytes to phytohemagglutin or concanavalin A. Chitotriose (a lysozyme inhibitor) caused a strong reduction in the inhibitory effects of the two lysozymes on the lymphocyte response to either lectin. HL and HEWL also were found to markedly inhibit the polyclonal B cell proliferation and differentiation induced by pokeweed mitogen and T cells. A less marked inhibition was also obtained when T cells, but not B cells, were pretreated with HL or HEWL. Again, as in the experiments with T cell mitogens, the effects were dose dependent and 5 micrograms/ml HL and 1 microgram/ml HEWL proved to be the most effective concentrations. The possible mechanisms by which lysozyme inhibits the lymphocyte response to mitogenic lectins are considered and discussed. The enzymatic activity seemed to perform an essential function, as shown by the loss of effect when the heat- or trypsin-inactivated lysozymes were used and by the fact that only the enzymatically active compound, among certain semisynthetic derivatives of HEWL, inhibited the lymphocyte response to the mitogens. However, the cationic properties of the lysozyme molecule appeared to be essential too, since enzymes with a similar specificity of action showed effects similar to those observed with HL or HEWL only when they carried a strong positive charge. It is suggested that lysozyme, which is naturally secreted by monocytes and macrophages, might interact with lymphocyte surface receptor sites and participate in the complex mononuclear phagocyte-lymphocyte interactions and in the modulation of lymphocyte activation.     

Acetaldehyde stimulation of the growth of Saccharomyces cerevisiae in the presence of inhibitors found inlignocellulose-to-ethanol fermentations

The addition of small quantities of acetaldehyde to fermentations containing inhibitory concentrations of furfural, acetate and other compounds typically present in lignocellulosic hydrolyzates significantly reduced the lag phase of yeast growth and stimulated ethanol production. Similar effects were observed when acetaldehyde (0.06 g l⁻¹) was added to fermentations of a birch wood hydrolyzate produced by steam/acid pretreatment. Acetaldehyde addition appears to have potential as a low-cost alternative (or adjunct) to current procedures for medium detoxification in lignocellulose-to-ethanol fermentations, particularly those in which high inhibitor concentrations are generated through recycling of the culture broth. Journal of Industrial Microbiology & Biotechnology (2000) 25, 104–108. Received 18 March 2000/ Accepted in revised form 02 June 2000     

Interaction of Saccharomyces cerevisiae with gold: toxicity and accumulation

This paper examines the effects of ionic gold on Saccharomyces cerevisiae, as determined by long-term (growth in gold-containing media) and short-term interactions (H⁺ efflux activity). An increasing gold concentration inhibited growth and at <0.2 mM Au, growth was not observed. Transmission electron microscopy revealed no differences in ultrastructure but fine electron dense particles were observed in unstained preparations from gold-containing medium. After glucose addition (to 10mM) to starved suspensions of S. cerevisiae, glucose-dependent reduction of external pH occurred as the cells extruded protons. In the presence of increasing gold concentrations, the lag time before proton extrusion did not change but the rate and duration decreased significantly with a marked influence on proton efflux rate being observed at 10 M. Extension of preincubation time of yeast cells in gold-containing medium resulted in a decreasing proton efflux rate and colloidal phase formation in the cell suspensions, the time between gold addition and the beginning of colloidal phase formation depending on the gold concentration used. Both Ca and Mg enhanced the inhibitory effect of gold on the yeast cells with Ca showing a stronger inhibitory effect than Mg.     

Reversal of primary embryonic induction by cold, and the "fixation" of induction in cells activated by lithium chloride

The effects of low temperature on embryonic induction were investigated. Presumptive epidermis cells of Rana pipiens were either pretreated with cold (4°C) and then treated with lithium chloride at the same temperature, or activated by lithium chloride at 22°C, then rapidly chilled to 4° and kept at this temperature for a period of time. Cultures were made from the treated cells. It was found that induction at low temperature could occur, although it did not proceed beyond determining neuralization. When, on the other hand, the cells were first induced with LiCl to the point at which they should have been determined to differentiate into melanophores, but were then rapidly chilled and kept in the cold for some time, the effects of induction were suppressed and the cells differentiated into ciliated epithelium. A 15–60 minute lag in time between induction and chilling (during which the cells were kept in the culture medium at room temperature) allowed neural induction to be gradually “fixed,” and a 120–180 minute lag was sufficient for melanphore induction to become insensitive to the subsequent treatment with cold. It is speculated from these results that induction occurs very slowly at low temperature and that the inductive stimulus activates a sequence of reactions, one or several of which are strongly repressed by the cold. In this case, when no products result from these reactions, the sequence is assumed to stop and possible feed-back mechanisms switch the whole system to its initial condition.     

Melafen action on the growth of cyanobacteria and green microalgae cultured in stress conditions

Melafen stimulating effect on cell growth of cyanobacteria Synechococcus sp. PCC 6301 cultures amounted to 30–45% at 1000 lx illumination. The melafen effect decreased when cell cultures were exposed at the illumination of the saturation range (4000 lx). Growth rate and biomass increase of Anabaena variabilis, as well as the observed melafen stimulating effect, were higher on nitrogen-free medium compared to a nitrogen-containing one by 20–25%. We conclude that melafen activates photosynthetic processes and, probably, stimulates fixation of the atmospheric nitrogen in the cells. Opposite to the stimulating effect of melafen, ions of the heavy metal Cd²⁺ inhibited both biomass increase and the average number of the cells in the cyanobacteria A. variabilis colonies. The melafen added to the medium together with the Cd²⁺ ions decreased their negative effect. The other heavy metal ions, Cu²⁺, inhibited the growth of the cyanobacteria Synechococcus sp. PCC 6301 and green microalgae Chlorella vulgaris but had a stimulation effect on carbohydrate excretion by the cell cultures. Again, the melafen decreased the toxic effect of Cu²⁺ in this case. We suppose that melafen has an antistress activity at heavy metal ions presence and reduces their toxic effect on growth of phototrophic microorganisms.     

CELL DEATH,SURVIVAL AND PROLIFERATION IN TETRAHYMENA THERMOPHILA. EFFECTS OF INSULIN,SODIUM NITROPRUSSIDE, 8-BROMO CYCLIC GMP,NG-METHYL-L-ARGININE AND METHYLENE BLUE

Cells of the ciliate Tetrahymena thermophila produce compounds that act as autocrine (paracrine) survival and/or growth factors. 8-Bromo cyclic GMP, sodium nitroprusside, hemin, protoporphyrin IX, human recombinant and bovine insulin were tested for their ability to substitute for the cell-produced factors and stimulate cell survival and proliferation. The cells were inoculated into conical flasks in a nutritionally complete, chemically defined medium at known cell densities from 5 to 5000cells/ml. In unsupplemented medium cells at 5 to 500cells/ml (‘low initial cell density cultures’) died within 8h, whereas cells at 1000 and 5000cells/ml (‘high initial cell density cultures’) proliferated with lag phases lasting for up to 4h. In the presence of insulin compounds, hemin, protoporphyrin IX, or 8-bromo cyclic GMP, cells also proliferated at all low initial cell densities. Sodium nitroprusside was effective over two separate concentration ranges: at the nanomolar levels as well at low pico- to femtomolar levels. At initial population densities of up to 50cells/ml the cells at both concentrations of sodium nitroprusside survived about 4-fold longer than the controls. At 500 initial cells/ml, cells at thehigh concentrations of sodium nitroprusside survived about 4-fold longer than those of the control cultures; they proliferated in the low concentrations of sodium nitroprusside. Concentrations of hemin, too low to have any effects on their own, had synergistic effects with sodium nitroprusside. N^G-methyl-L-arginine inhibited proliferation at high initial cell densities. This inhibitory action was reduced by high concentrations of L-arginine, protoporphyrin IX, sodium nitroprusside, or 8-bromo cGMP, but not by insulin. Methylene blue inhibited cell proliferation at high initial cell densities. This inhibition was circumvented by addition of 8-bromo cGMP. The findings that insulin-related material may be released from Tetrahymena and that insulin and sodium nitroprusside increase intracellular cGMP in these cells are discussed in relation to the presented results. Together these observations suggest that cGMP is responsible for supporting cell survival in Tetrahymena and switching the cells into their proliferative mode, and that cell-produced signal molecules and insulin stimulate an NO-dependent guanylate cyclase into producing cGMP.     

Effect of Inhibitors of Bacterial Cell Wall Synthesis on the Growth of Anabaena variabilis,a Blue-green Alga

The effects of penicillins and several other antibiotics (vancomycin, ristocetin, bacitracin, novobiocin, and d-cycloserine), all known as inhibitors of bacterial cell wall synthesis, were tested on the growth of Anabaena variabilis, a strain of blue-green alga.

All the antibiotics tested inhibited the growth of Anabaena variabilis at concentrations ranging from 10 µg to 1 mg per ml. However, penicillins and the other antibiotics tested did not inhibit growth of strains of green algae.