Interactions between the unicellular red alga Rhodella reticulata (Rhodophyta) and contaminated bacteria

AIMS: To define the role of the bacterial strains LR1 and LR3 in the Rhodella cell destruction caused by Cytophaga sp.LR2. METHODS AND RESULTS: The bacteria were obtained from algal culture with destruction. They were isolated in pure culture and tested for biochemical activities using Polymicrotest. The ability of bacteria to degrade and utilize the algal polysaccharide was investigated. The bacteria were grown in a media containing Rhodella polysaccharide as a sole carbon source. The level of the reducing sugars in the culture media was determined. Scanning electron microscopy (SEM) was used to define the location of bacteria in extensively and intensively cultivated Rhodella reticulata previously infected by Cytophaga sp. LR2. CONCLUSIONS: The lysis of Rhodella reticulata cells is due to the joint action of the three bacterial strains with the former pathogen Cytophaga sp. LR2 playing the main role. The accumulation of the polysaccharide and the excreted metabolites of the strains LR1 and LR3 stimulated the development of Cytophaga sp. LR2. The adaptation of the strain to particular conditions of alga cultivation and the utilization of polysaccharide as a sole carbon source supported its stable growth in alga suspension and destruction of Rhodella cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The predominance of Cytophaga sp. LR2 over the two other contaminants and the lysis of Rhodella reticulata cells resulted from the ability of the bacterium to attach to the algal polysaccharide sheath. The formation of slime and extrusions facilitated the phenomenon of bacterial adhesion to the algal surface as well as the formation of colonial alga - bacterial spherules. The sedimentation of these aggregates decreased the ability of the algal strain to photosynthesize, led to the lysis of the cells and finally caused the death of Rhodella.     

BIOTIC RELATIONSHIPS BETWEEN SOIL ALGAE AND OTHER MICROORGANISMS

Parker , Bruce C., and Harold C. Bold . (U. Texas, Austin.) Biotic relationships between soil algae and other microorganisms. Amer. Jour. Bot. 48(2): 185–197. Illus. 1961.—A study was conducted of biotic relationships between various algae and other microorganisms isolated from a sample of Texas soil. From 143 two-membered combinations of organisms tested in soil-water cultures, 3 were selected for detailed studies of the nature of the causal mechanisms of the associative effects. These were: (1) an association between a species of Bracteacoccus (Br. A-20) and a heterotrophic bacterium (B-6); (2) an association between a species of Chlamydomonas (Ch. 10) and a species of Streptomyces (Act-1); and (3) an association between a blue-green alga, Phormidium sp. (Ph. 14), and an as-yet-unidentified fungus (F-2). In the first association, the heterotrophic bacterium increased growth of Bracteacoccus up to 20 times in soil-water culture; the chief cause of stimulation was shown by a series of experiments to be the decomposition by the bacterium of complex nitrogenous substrates in the soil resulting in the release of simplified products which were available to the alga as a nitrogen source. In soil-water culture, Streptomyces (Act-1) enhanced the growth and motility of Chlamydomonas (Ch. 10) and induced akinetogenesis in the alga, while the actinomycete itself was stimulated in growth and production of conidia. The mutual stimulation was shown to be caused, in part, by carbon dioxide-oxygen interchange between the organisms. Motility of Ch. 10 was enhanced by a decrease in nitrogen as a result of the growth of Act-1 and by the ability of Act-1 to decompose and assimilate the extracellular polysaccharide of the alga. The assimilation of the extracellular polysaccharide by the actinomycete promoted its growth and conidia production. Initiation of akinetogenesis in Chlamydomonas occurred exclusively in close association with the filaments of Streptomyces in soil-water cultures, and only when the concentration of available nitrogen dropped below a critical level. The akinetogenic factor has not yielded to isolation and identification, but there is some suggestion that it may be an antibiotic substance. Phormidium (Ph. 14) was frequently antagonized and annihilated by fungus 2 in the soil-water medium. Attempts to extract growth inhibitors from filtrates of the fungal medium were unsuccessful. Indirect evidence suggested that perhaps the consumption of extracellular polysaccharide with concomitant release of organic acid by the fungus might be the factor inhibiting growth of Phormidium. Attempts to confirm experimentally the ecological significance for these biotic relationships are reported.     

Light quality affects morphology and polysaccharide yield and composition of Gelidium sesquipedale (Rhodophyceae)

Morphology and polysaccharide characterization of Gelidium sesquipedale (Clem.) Bornet et Thuret were studied in cultures grown under various light qualities. White light (WL), blue light (BL) and red light (RL) (all at photon fluence rate of 40 μmol m-2 s-1) were used for the study of morphological characteristics, and in addition yellow light (YL) for polysaccharide characterization. RL and BL induced a proliferating growth, which resulted in bushy plants under RL. Cortical cells of BL-grown plants were smaller and presented a higher density per unit area, whereas those of WL- and RL-grown alga were larger. Medullary cells followed the inverse pattern. Light quality also affected polysaccharide yield and composition, with the yield being higher under BL, RL or YL than WL. Most of the polysaccharide was extracted in distilled water at 100 °C, while a low amount was solubilized at 22 °C and 120 °C. Extracts from BL-grown alga presented the highest galactan content. The starch concentration was lower in extracts from RL-, BL- and YL-cultivated alga than in those from the initial plants. The degree of substitution with methoxyl groups and precursor was very low in all the agar fractions, but fractions extracted from BL- and WL-grown alga were more substituted by precursor. The highest sulfate content was reached under BL (about 9% w/w) and the highest 2-O-methyl-3,6-anhydro-L-galactose and 6-O-methyl-D-galactose content were found in extracts from alga grown under YL. This revised version was published online in June 2006 with corrections to the Cover Date.     

BRYOPHYTE LEAF EPIFLORA: AN SEM AND TEM STUDY OF POLYTRICHUM COMMUNE HEDW.

Epiphytes on leaves of Polytrichum commune Hedw. were observed and studied with SEM and TEM. A unicellular green alga, a fungus, and a rod-shaped bacterium were present on both leaf surfaces. Culturing experiments led to the identification of the alga as Chlorococcum sp. and the fungus as Trichoderma viride Lk. ex Fr. (Deuteromycetes). No attempt was made to culture and identify the bacterium. Terminal cells of the leaf lamellae form a ventral pseudo-epidermis which provides a microhabitat for the epiphytes. Fungal hyphae were seen intracellularly as well as penetrating cell walls between lamellar cells. The alga and bacterium were not observed intracellularly. Chlorococcum sp. displayed a distinctive wall morphology which consisted of short (0.3–0.5 μm), blunt papillae. The rod-shaped bacterium may be similar to the Agrobacterium-like isolates reported recently from Polytrichum commune. Host-epiphyte associations relative to potential beneficial and/or pathological aspects of growth require further investigation.     

Comparative study of carrageenans from reproductive and sterile forms of <Emphasis Type="Italic">Tichocarpus crinitus</Emphasis> (Gmel.) Rupr (Rhodophyta,Tichocarpaceae)

A comparative study of the structure and properties of the sulfated polysaccharides (carrageenans) isolated from the vegetative and reproductive forms of the red alga Tichocarpus crinitus was performed. The polysaccharides were separated into the gelling (KCl-insoluble) and non-gelling (KCl-soluble) fractions by precipitation with 4% KCl. The total content of polysaccharides extracted from the reproductive form of the alga was 1.8-fold more than that extracted from the vegetative form, and in the first case, the gelling polysaccharides mostly accumulated. The gelling polysaccharides from the vegetative form have the highest molecular weight (354 kD). According to the results of FT-IR and 13C-NMR spectroscopy, the gelling polysaccharide fractions from both forms are kappa/beta carrageenans. The differences concern the content of the kappa- and beta-disaccharide units and the presence of a small content of the sulfated disaccharide segments (precursors of the kappa-carrageenans) in the polysaccharide from the reproductive form of the alga. The non-gelling polysaccharide fractions from both forms of the plant are mixtures of sulfated galactans with a low content of 3,6-anhydrogalactose.     

Production of Polysaccharide from Starch and Identification of Constituent Sugars of the Preparation

A bacterium which belongs to Achromobacter sp. was isolated and named as Achromobacter mucosum nov. sp. Starch and dextrin were essential carbon sources to produce a polysaccharide effectively by the bacterium. Maltotriose was as effective as starch for the production of the polysaccharide, and glucose, maltose, isomaltose and panose were little effective. This indicates that the bacterium requires a definite configuration of carbon source to produce the polysaccharide effectively. The dry powder of the polysaccharide was prepared from 10 liters of broth in the yield of 34.9 g. Glucose, galactose, mannose and uronic acid were confirmed as the constituent sugars of the polysaccharide and it was most probable that the uronic acid was d-glucuronic acid.     

Effect of Bacteria Associated with the Green Alga Ulva reticulata on Marine Micro- and Macrofouling

The green alga Ulva reticulata (Forsskal) is often free from biofouling in Hong Kong waters. An early study indicated that bioactive substances from this alga inhibit settlement of the polychaete Hydroides elegans (Haswell). It is also predicted that epibiotic bacteria protect this alga from micro- and macrofouling. In this study, bacterial strains from the surface of U. reticulata were isolated and their inhibitive activities on micro- and macrofouling assayed. The strains were identified by 16S rRNA analysis as belonging to the genera Alteromonas , Pseudoalteromonas and Vibrio . There was no significant effect of these strains or their extracts (aqueous and ethanol) on the growth of five Vibrio strains isolated from natural biofilm. Two bacterial strains ( Alteromonas sp. and Vibrio sp. 3) were non-toxic to the benthic diatom Nitzschia paleacea (Grunow) while the other five strains caused a low level of mortality. No one bacterial strain was toxic to the larvae of H. elegans . Aqueous extract of one of the isolated bacterial species, i.e. Vibrio sp. 2, significantly ( p <0.00001) inhibited the settlement and metamorphosis of H. elegans larvae. The putative antifouling compounds have a molecular weight of >100 kD. On the other hand, biofilm of Pseudoalteromonas sp. 2 and aqueous extract of Vibrio sp. 2 suppressed the settlement of larvae induced by 3-isobutyl-1-methylxanthine (IBMX). Other epibiotic bacteria and their extracts had neither inhibitive nor inductive effects on larval settlement of H. elegans . The results indicate that the antifouling mechanism of U. reticulata may be dependent not only on materials from the macroalga itself but also on the epibiotic bacteria on the algal surface.     

Degradation of unsaturated polysaccharide derived from acidic polysaccharide of Fusarium sp. M7-1 by a bacterium isolated from soil

A soil bacterium, strain no. 19, capable of using unsaturated polysaccharide derived from acidic polysaccharide of Fusarium sp. M7-1 as a sole source of carbon was isolated. The bacterium degraded about 70% of the total sugar content. Results from analysis of the degraded polysaccharide showed that the bacterium degraded the β(1→6) galactofuranoside linkage as well as the unsaturated glucuronic acid residues linked to the galactofuranoside residues via the α(1→2) linkage.     

Characterization of Rhizobium sp. Q32 Isolated from Weathered Rocks and its Role in Silicate Mineral Weathering

One mineral-solubilizing strain was isolated from weathered potassic trachyte surfaces and identified as Rhizobium sp. Q32 based on the 16S rRNA gene sequence analysis. The mineral (feldspar and biotite) dissolution potential and the physiological characteristics of the bacterium were investigated. Laboratory mineral dissolution experiments indicated that inoculation with the live bacterium significantly increased feldspar and biotite dissolution by a factor of 1.2–4.7 for Si and 1.2–1.5 for K in comparison with the dead bacterium inoculated controls. In addition, extracellular polysaccharide production by the bacterium increased with time but the bacterium produced small pH changes (6.0–6.5) in the course of mineral dissolution experiment. The bacterium was found to produce siderophores and have the characteristics of acid or alkali and salt tolerance and temperature resistance. The result suggested that feldspar and biotite dissolution may be mainly caused by extracellular polysaccharide and/or siderophores produced by the bacterium.     

Influence of physiological factors on the lysis effect of Cytophaga on the red microalga Rhodella reticulata

The influence of different factors on the lysis of the red microalga, Rhodella reticulata, by Cytophaga sp. LR2 was studied. The pathogenic bacterial strain was more resistant than the alga to the physiological parameters studied, which assured long-term survival of bacteria in algal cultures. Cytophaga sp. LR2 infected R. reticulata at temperatures between 15 and 30 degrees C, in the illuminated as well as the non-illuminated cultures, at pH values between 5.0 and 9.0, and in the presence of NaCl and CaCl2 in the culture medium. SEM showed a different morphology of the bacteria in algal cultures from those of axenic cultures of Cytophaga. Observations of specific associations between algal and bacterial cells revealed that the role of the slime extrusions on the bacterial surface was attachment of Cytophaga to algal cells, and that their clumping leads to rapid lysis.     

A sulfated fucan from the brown alga Laminaria cichorioides has mainly heparin cofactor II-dependent anticoagulant activity

The major acidic polysaccharide from the brown alga Laminaria cichorioides is a complex and heterogeneous sulfated fucan. Its preponderant structure is a 2,3-disulfated, 4-linked alpha-fucose unit. The purified polysaccharide has a potent anticoagulant activity, as estimated by APTT assay ( approximately 40 IU/mg), which is mainly mediated by thrombin inhibition by heparin cofactor II. It also accelerates thrombin and factor Xa inhibition by antithrombin but at a lower potency. Sulfated fucan from L. cichorioides is a promising anticoagulant polysaccharide and a possible alternative for an antithrombotic compound due to its preferential heparin cofactor II-dependent activity.     

中华蜜蜂在滇山茶上的访花行为研究

对中华蜜蜂在滇山茶上的访花行为的研究表明,中华蜜蜂全天在滇山茶上的访花过程呈现出单峰型的节律,高峰在13:00～14:00.在31朵观察的样花上,中华蜜蜂全天共到访613次,访花627次,对花的连续访问率为2.23％,每朵花平均被访20.23次,每次到访花停留时间均值为10.23 s,全天在样花上总的累计停留时间为6 270 s,单花累计访问时间达到202.26 s.蜜蜂全天访花累计时间占全部访花昆虫总访花的累计停留时间的比例达到97.80％.在滇山茶访花昆虫中,中华蜜蜂具有明显的优势度,是绝对的优势种.对滇山茶花蜜的研究表明,其单花1天的花蜜分泌量平均为69.5 μL(n=100);花蜜含糖量(124.95～200.64) mg/mL.花蜜成分以多糖为主,占66％以上,此外还含有单糖,占1.9％～33％,其他未知名的组分占16％～22％,部分花蜜中含有山梨醇,占11％.对滇山茶花的挥发物分析结果表明,总的挥发物有59种,萜烯类、酯类和醇类是滇山茶花的主要挥发物成分,分别占46.468％、27.035％和21.609％.     

Role of Hydrophobicity in Adhesion of the Dissimilatory Fe(III)-Reducing Bacterium Shewanella alga to Amorphous Fe(III) Oxide

The mechanisms by which the dissimilatory Fe(III)-reducing bacterium Shewanella alga adheres to amorphous Fe(III) oxide were examined through comparative analysis of S. alga BrY and an adhesion-deficient strain of this species, S. alga RAD20. Approximately 100% of S. alga BrY cells typically adhered to amorphous Fe(III) oxide, while less than 50% of S. alga RAD20 cells adhered. Bulk chemical analysis, isoelectric point analysis, and cell surface analysis by time-of-flight secondary-ion mass spectrometry and electron spectroscopy for chemical analysis demonstrated that the surfaces of S. alga BrY cells were predominantly protein but that the surfaces of S. alga RAD20 cells were predominantly exopolysaccharide. Physicochemical analyses and hydrophobic interaction assays demonstrated that S. alga BrY cells were more hydrophobic than S. alga RAD20 cells. This study represents the first quantitative analysis of the adhesion of a dissimilatory Fe(III)-reducing bacterium to amorphous Fe(III) oxide, and the results collectively suggest that hydrophobic interactions are a factor in controlling the adhesion of this bacterium to amorphous Fe(III) oxide. Despite having a reduced ability to adhere, S. alga RAD20 reduced Fe(III) oxide at a rate identical to that of S. alga BrY. This result contrasts with results of previous studies by demonstrating that irreversible cell adhesion is not requisite for microbial reduction of amorphous Fe(III) oxide. These results suggest that the interaction between dissimilatory Fe(III)-reducing bacteria and amorphous Fe(III) oxide is more complex than previously believed.     

Chemical characterization and radioprotective effect of polysaccharide from <Emphasis Type="Italic">Monostroma angicava</Emphasis> (Chlorophyta)

Polysaccharide from the green alga Monostroma angicava was extracted with boiling water and was purified by ion-exchange and size-exclusion column chromatography. The radioprotective effect of the polysaccharide was investigated in mice. The results show that polysaccharide from M. angicava has a different chemical composition to other Chlorophyta having a high rhamnose – containing sulfated polysaccharide. The sulfate ester content was estimated to be 21.8%. When the polysaccharide was applied to BALB/c mice following whole-body X-ray irradiation the counts of leukocytes, thrombocytes and erythrocytes recovered more rapidly in the polysaccharide treated mice after irradiation. In the irradiated mice, the polysaccharide significantly increased the spleen index, natural killer cytostatic activity and the transformation response of splenic lymphocytes. The present observations suggest that polysaccharide from M. angicava led to leukocytogensis and hematopoetic activation in mice after irradiation and that the biological response might be caused by immune activation.     

Extracellular Polysaccharide Formed by Pseudomonas hydrogenovora in Autotrophic Culture and Its Physiological Activities

Gram negative hydrogen bacterium Pseudomonas hydrogenovora was found to excrete an anthrone-H₂SO₄ positive viscous polysaccharide. About 12 g/liter of the polysaccharide was produced autotrophically on gaseous hydrogen at the stationary phase of growth. Biosynthesis of the polysaccharide occurred under nitrogen-deficient condition. Its elementary composition was C: 39.29%, H: 6.23%, O: 49.67%, N: 0.21% and ash 4.6%. The polysaccharide contained galactose, glucose, mannose and rhamnose as its main components. The polysaccharide had anti-tobacco mosaic virus and anti-tumor activities.     

Isolation and characterization of a sulfated polysaccharide from the brown alga Sargassum patens and determination of its anti-herpes activity.

Bioactivity-guided fractionation of the hot water extract from the brown alga Sargassun patens led to the isolation of a polysaccharide as an antiviral component against herpes simplex viruses which are the cause of cold sores (HSV-1) and genital herpes (HSV-2). The polysaccharide contained a sulfur group that could be present as a sulfate ester. It is thus a sulfated polysaccharide with a molecular mass of about 424 kDa, and is designated SP-2a. Gas chromatographic assay showed that the polysaccharide consisted of fucose, galactose, mannose, xylose, glucose, and galactosamine. The fucose is the major constituent sugar (35.3%), followed by galactose (18.4%). The 50% effective concentration (EC50) against HSV-2, HSV-1, and HSV-1 acyclovir resistant strain was 1.3, 5.5, and 4.1 microg/mL, respectively. The 50% cytotoxic concentration (CC50) of SP-2a on the growth of normal Vero cell line was more than 4000 microg/mL. Therefore SP-2a of S. patens may be a potent agent for treating HSV infections.     

Levan production byBacillus polymyxa

Summary A levan-producing bacterium was isolated from soils and its characteristics for polysaccharide synthesis were studied. A series of enrichment and plating techniques enabled the isolation of a levan-producing bacterium from closely related contaminants. Cultural and physiological characteristics of the isolate identified the organism an a strain ofBacillus polymyxa. The organism produced about 40 g extracellular polysaccharide per liter of sucrose medium, which was about three times more yield than levan obtained from known levan producers. The highest amount of polysaccharide was on a 8% sucrose medium. Hydrolysis of the product showed that the polysaccharide consisted entirely ofd-fructose, and¹³C.n.m.r. spectra confirmed that the product was levan, a fructose polymer linked by B-(26) fructofuranosyl linkage.     

Production of extracellular polysaccharides by Lactobacillus delbrueckii ssp. bulgaricus NCFB 2772 grown in a chemically defined medium

Lactobacillus delbrueckii ssp. bulgaricus NCFB 2772 produced an extracellular polysaccharide when grown in a chemically defined medium with glucose or lactose as the substrate carbohydrate. The isolated extracellular polysaccharide had a sugar composition of glucose, galactose and rhamnose in a ratio of 1:6.8:0.7. The production of extracellular polysaccharides increased at higher temperatures, but the bacterium rapidly lost its polysaccharide producing ability at 47°C. Production of polysaccharides was growth-related: no polysaccharide production was found after growth had ceased. An excess carbohydrate did not result in increased polysaccharide production.     

GROWTH AND POLYSACCHARIDE PRODUCTION BY THE GREEN ALGA CHLAMYDOMONAS MEXICANA (CHLOROPHYCEAE) ON SOIL1

The polysaccharide producing soil alga Chlamydomonas mexicana Lewin was grown on soil surfaces in a growth chamber, with cell number and total polysaccharide measured weekly. Cell growth was pronounced, reaching an excess of 10⁷ cells · cm^?2 within one week. Polysaccharide production was also pronounced, with similar amounts of polysaccharide synthesized whether or not the cells were continuously given nutrients. Polysaccharide synthesis increased once the cells slowed in their growth, as in previous work with the cells in liquid medium.     

A yellow polysaccharide-producing bacterium with unusual characteristics

Summary A polysaccharide-producing Gram-negative bacterium was isolated from a sample of hay. It grew best on nitrate-containing media with sucrose as carbon source; the colony form was highly unusual. No polysaccharide was formed on glucose. A spontaneous mutant producing the same polysaccharide on both sucrose- and glucose-containing media was isolated. The polysaccharide has been characterized as an acid heteropolymer containing D-galacturonic acid, D-glucose, D-galactose and D-mannose in the approximate molar ratio 1:1:3:1, together with about 2 mol of acetate. The polysaccharide in aqueous solution was highly viscous with pseudoplastic characteristics.