Salinity tolerances and ecological aspects of some fungi collected from fresh-water,estuarine and marine habitats

Summary Twenty species of Fungi imperfecti, including strains from freshwater, estuarine and marine habitats in the environs of the middle and lower Neretve River, Yugoslavia, were collected, purified and identified. Strains of nine species from fresh, brackish, and sea water were grown on various NaCl concentrations. These fungi exhibited a wide amplitude of salt tolerance. In general, the strains from fresh water grew best at or were able to tolerate the lower NaCl concentrations. The sea-water strains grew best at and could tolerate the higher NaCl concentrations. Brackish-water strains were found to be more like strains isolated from fresh water than from sea water. Brackish-water strains were able to grow well in zero or low concentrations of NaCl, but they could not do as well at the higher NaCl concentrations. The results of the laboratory experiments with NaCl concentrations support the findings in nature.All twenty species of these fungi were tested for their ability to decompose lignin, utilize phosphate and degrade lecithin. Five species gave a positive reaction on gallic acid medium; five species were able to utilize lecithin; and seven species were active in utilizing tribasic calcium phosphate.     

D- and l-amino acid concentrations in culture broth of Lactobacillus are highly dependent on the phylogenetic group of Lactobacillus

d-amino acids produced by Lactobacillus are thought to contribute to the taste quality and health functions; however, no studies have comprehensively evaluated the concentrations of the D- and L-forms of amino acids separately in individual Lactobacillus strains. To gain insight into amino acid concentrations in Lactobacillus, we evaluated amino acid concentrations in culture broth of Lactobacillus separately for the D- and L-forms. Lactobacillus strains were cultured in culture broth, and the amino acid concentrations in supernatant were assessed. The amino acid concentrations obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) were subjected to cluster analysis based on Bray-Curtis distance with Ward's minimum variance method. In the analysis of amino acid concentrations under culture with different monosaccharides, the distances among strains cultured with the same monosaccharide were significantly greater than those among cultures of the same strain under different monosaccharides (p < 0.01). The cluster analysis of amino acid concentrations under culture with the same monosaccharide suggested that strains belonging to the same phylogenetic group of Lactobacillus exhibited similar concentrations of amino acids. Data analyses of 70 strains belonging to 17 Lactobacillus taxa indicated that the concentrations of amino acids were highly dependent on the phylogenetic group of Lactobacillus and that the group differences in amino acid concentration were strongly driven by differences in l-serine and d-alanine concentrations. Our results indicate that it is important to evaluate D- and l-amino acids separately when evaluating variations in amino acid concentrations. Because d-alanine has the potential to affect taste quality, the results of this study may provide insight into the taste quality of fermented food produced by Lactobacillus.     

Selective control of Microcystis using an amino acid – a laboratory assay

An amino acid (L-lysine) was screened against eighteen strains ofCyanophyceae, Bacillariophyceae and Chlorophyceae. OnlyMicrocystis strains (Cyanophyceae) were sensitive tolysine; other strains were not affected. Cells of sevenMicrocystis strains (10⁶ cellsmL^–1) were completely killed within 48h by lysine at concentrations between 0.6 and 5.0 mgL^–1. Two Microcystis strains wereinhibited by 92 and 98 %. Similar results were obtained when lysine malonateandlysine copper were used as algicides. Microcystis specieswere killed by lysine malonate at concentrations between 0.6 and 5 mgL^–1, and by lysine copper at concentrations between 0.5and 20 mg L^–1.     

Selection of Wine Yeasts for Growth and Fermentation in the Presence of Ethanol and Sucrose

To optimize the conversion of carbohydrates to ethanol, strains of several Saccharomyces species were examined for the ability to grow and ferment in a range of sucrose and ethanol concentrations. A total of 632 wine yeasts, most of them isolated from wineries in Andalusia and Extremadura, southwestern Spain, were subjected to screening and selection. Growth and fermentative capacity in different ethanol and sucrose concentrations varied from one strain to another. There was no correlation between growth and fermentative capacity. The best 35 strains grew in 15% ethanol and fermented in 18% ethanol. Ethanol accumulated, although at a reduced rate, after the cells stopped growing. Most yeast strains were highly fermentative in 50% sucrose. Some of them effectively utilized the carbohydrates of the culture, yielding final ethanol concentrations of > 14%. Of the 35 selected strains, 16 were promising for genetic analysis and breeding because of their capacity to sporulate. These strains were homothallic, and their spores were viable. The meiotic products analyzed so far were also homothallic.     

Antibacterial Activity of a New Cephalosporin, Cefotaxime

The antibacterial activity of a new cephalosporin derivative, cefotaxime (HR 756), was determined. The antibiotic was active at low concentrations against R⁺ and R^- strains of Gram negative bacteria, including two out of three strains of Serratia marcescens. In general higher concentrations were needed to inhibit growth of Pseudomonas aeruginosa. Low concentrations induced elongation of cells in circumstances conducive to active growth; higher concentrations caused lysis in some strains. Cefotaxime was more stable than cephaloridine, cephalothin, cephalexin, cefoxitin and cefuroxime to various β-lactamases.     

Influence of Serratia marcescens Pigmentation on Cell Concentrations in Aerosols Produced by Bursting Bubbles

For eight strains of Serratia marcescens, increased cell concentrations were found in aerosols produced from bursting bubbles, with concentrations ranging from a maximum of ca. 80 times the bulk concentration for pigmented strains 4180, 933, and 274 to a minimum approximately equal to the bulk concentration for nonpigmented strain 8100. The increased cell concentration in the aerosol was suppressed when pigmented strains were grown at 37°C, a temperature at which the pigment prodigiosin is not synthesized, resulting in lower concentrations similar to those of nonpigmented strains. Strains that produce higher concentrations of prodigiosin after 1, 2, 4, and 8 days of growth show increasing concentrations in bubble-produced drops; duplicate cultures grown at 37°C did not show any increases. In four concurrent experiments, cells starved for 24 h showed greater concentrations than nonstarved cells for chromogenic strain NIMA, whereas for nonchromogenic strain WF, starved cells showed greater concentrations in three cases and a decreased concentration in the fourth. Bacterial concentrations in aerosol drops from bursting bubbles appear to be predominantly influenced by the surface condition of the bacterial cell.     

Variation of Microcystin Content of Cyanobacterial Blooms and Isolated Strains in Lake Grand-Lieu (France)

Abstract Cyanobacterial blooms were sampled at five locations in Lake Grand-Lieu on seven different occasions during May–October 1994. Strains of Microcystis aeruginosa and Anabaena circinalis were isolated from the samples. Microcystins were detected in freeze-dried field samples and the isolated strains by HPLC. The toxins were present in the blooms sampled between June and October. The microcystin content in the blooms varied with site and time, from undetectable concentrations to 0.23 mg g⁻¹. The highest concentrations of microcystin were found in blooms sampled in September. Microcystin-LR and microcystins with retention times close to the retention time of [Dha⁷]microcystin-RR (probably varieties of microcystin-RR) were found in the field samples. Sixteen of the 98 isolated M. aeruginosa strains and 2 of the 24 A. circinalis strains produced microcystins. The total amount of microcystins varied from undetectable concentrations to 5.06 mg g⁻¹ in the M. aeruginosa isolates, and from undetectable concentrations to 1.86 mg g⁻¹ in the A. circinalis strains. Microcystin-LR was the main toxin found in strains of M. aeruginosa, but was not present in strains of A. circinalis. Both microcystin-producing strains and strains that did not produce microcystin coexisted in the bloom samples. Received: 23 January 1997; Accepted: 25 March 1997     

A new method of preparation of pyocyanin and demonstration of an unusual bacterial sensitivity.

Pyocyanin was prepared in 60% yield from phenazine methoxysulfate by a photooxidation procedure and purification by silica gel chromatography. Monitoring was performed by thin-layer chromatography. Approximately 50% of clinical Pseudomonas aeruginosa isolates were found to produce pyocyanin at 37°C. Among Proteus strains, P. morganii strains were sensitive to concentrations of pyocyanin 16 to 64 times lower than concentrations that inhibited the growth of P. mirabilis and P. vulgaris strains.     

Photoinactivation and long-term photosensitivity of diploid and disomic strains of the smut fungusUstilago violacea

Diploid and disomic strains ofUstilago violacea, containing high cellular concentrations of cytochrome c and various cellular concentrations of carotenes, were subjected to high levels of radiation from incandescent, fluorescent, and UV sources to characterize inactivation kinetics. Additionally, these strains were grown under high and low levels of radiation from the fluorescent and incandescent sources, to characterize their long-term resistance to photoinactivation by visible light with different spectral qualities. Survival kinetics in response to incandescent radiation were characterized by either an exponential decay for the low carotene strain, or an initial loss of viability, with recovery and a subsequent gradual loss of viability for the strains containing high carotene concentrations. Long-term colony survival was suppressed at the high incandescent level for the low carotene strain only. Long-term incandescent exposure did not affect the other strains. Survival kinetics in response to fluorescent and UV radiation were characterized as exponential decays for all strains, however, the exposure time leading to 90% loss in viability LD₉₀ was quantitatively related to carotene content in the form of a power function, where LD₉₀ increased as the total carotene content per cell. We also determined that the ratio of colored carotenes to cytochrome c in the cells was quantitatively related to the rate of viability loss during UV exposure. In all cases, including the long-term fluorescent experiment, the strains containing higher concentrations of carotenes were more resistant to destruction by the different radiation sources.     

Glutathione status and sensitivity to GSH-reacting compounds of Escherichia coli strains deficient in glutathione metabolism and/or catalase activity

The intracellular concentrations of total glutathione, GSSG and protein · S-SG, the total excreted glutathione concentration, and the susceptibility towards GSH-reacting compounds were assayed in strains of Escherichia coli deficient in biosynthesis and/or reduction of glutathione. A deficiency in glutathione reductase displaced the glutathione status towards the oxidized forms. This displacement was more clearly appreciated in strains additionally deficient in glutathione biosynthesis. A deficiency in catalase activity also produced an increase in the oxidation of glutathione. The most severe changes were observed in the concentrations of protein-glutathione mixed disulfides and in the amount of glutathione excreted to the medium. Increased sensitivities towards compounds known to interact with cellular GSH were observed in glutathione reductase deficient strains, although these effects were enhanced in strains additionally deficient in GSH biosynthesis     

Further investigations on the concentration of marine bacteriophages in the water around Helgoland,with reference to the phage-host systems encountered

Between April 3 and September 24, 1991, the concentrations of bacteriophages infecting bacterial strains, isolated in 1990 and during this investigations, were determined in 35 samples of seawater taken at station ‘Kabeltonne’ adjacent to Helgoland. Similar to the findings of 1990, phage concentrations of several hundred plaque forming units (PFU) ml⁻¹ were observed with a number of indicator strains, the maximum concentration being at least 1.5×10³ PFU ml⁻¹. These high concentrations lasted for only a few days, generally decreasing at rates between 0.6 and 0.9 day⁻¹. Phage concentrations of 0 to 2 PFU ml⁻¹ were found to be predominant until the end of June, occasionally attaining 5 PFU ml⁻¹. From July through September, when high phage concentrations were observed with some indicator strains, between 0 and 10 PFU ml⁻¹ were found in the majority of tests. As revealed by a final phage-host cross-reaction test, the greater part of 138 indicator bacteria is genetically related, and almost half of the 200 phage strains tested are propagated only by their original indicator bacterium. The possible importance of mutational events for the maintenance of phage-host systems in nature is discussed.     

Some technological properties of Penicillium roqueforti strains isolated from a home-made blue cheese

Nine strains of Penicillium roqueforti isolated from a traditional Spanish blue cheese (Valdeón cheese) along with two commercial strains were investigated for their ability to grow at different concentrations of salt and at different temperatures as well as for their proteolytic and lipolytic activities. Low concentrations of salt (1-3%) were stimulating for all the strains, with 1% salt being the concentration with the highest stimulating effect in nearly all. The rate of growth at 10°C was 2-3 times lower than at 25°C, the optimum temperature for the species. None of the strains, including the commercial cultures, showed proteolytic activity on casein agar, while all of them were lipolytic on tributyrin agar.     

分离自黄瓜的多主棒孢霉不同表型菌株对杀菌剂的敏感性

【目的】通过对分离自黄瓜的多主棒孢霉不同表型菌株适宜生长温度、产孢量等表型特征和对8种杀菌剂的敏感性研究,为多主棒孢霉侵染引起的黄瓜叶斑病和茎疫病的化学防治提供技术支持。【方法】采用温度梯度法测定病菌适宜生长温度;采用PDA培养基25°C黑暗培养5 d和21 d,计测不同表型菌株单位面积产孢量;采用含药平板法测定不同表型菌株对8种杀菌剂的敏感性。【结果】分离自黄瓜的多主棒孢霉不同表型菌株适宜生长温度为25-30°C;在PDA平板25°C黑暗培养5 d,气生菌丝稀少型菌株cu-4、cu-5即大量产孢,产孢量明显多于气生菌丝丰茂型菌株;在试验浓度下,5个试验菌株对8种杀菌剂的敏感性依次为:代森锰锌氟硅唑戊唑醇苯醚甲环唑速克灵百菌清嘧菌酯多菌灵。【结论】分离自黄瓜的多主棒孢霉不同表型菌株在适宜生长温度、菌丝生长速度、产孢量及对杀菌剂的敏感性等方面均存在差异。在试验浓度下,供试菌株对多菌灵和嘧菌酯的敏感性极低(抑制率40%),这2种杀菌剂已失去对该地区黄瓜褐斑病的防控作用。     

Bacterial Degraders of Polycyclic Aromatic Hydrocarbons Isolated from Salt-Contaminated Soils and Bottom Sediments in Salt Mining Areas

Fifteen bacterial strains capable of utilizing naphthalene, phenanthrene, and biphenyl as the sole sources of carbon and energy were isolated from soils and bottom sediments contaminated with waste products generated by chemical- and salt-producing plants. Based on cultural, morphological, and chemotaxonomic characteristics, ten of these strains were identified as belonging to the genera Rhodococcus, Arthrobacter, Bacillus, and Pseudomonas. All ten strains were found to be halotolerant bacteria capable of growing in nutrient-rich media at NaCl concentrations of 1–1.5 M. With naphthalene as the sole source of carbon and energy, the strains could grow in a mineral medium with 1 M NaCl. Apart from being able to grow on naphthalene, six of the ten strains were able to grow on phenanthrene; three strains, on biphenyl; three strains, on octane; and one strain, on phenol. All of the strains were plasmid-bearing. The plasmids of the Pseudomonas sp. strains SN11, SN101, and G51 are conjugative, contain genes responsible for the degradation of naphthalene and salicylate, and are characterized by the same restriction fragment maps. The transconjugants that gained the plasmid from strain SN11 acquired the ability to grow at elevated NaCl concentrations. Microbial associations isolated from the same samples were able to grow at a NaCl concentration of 2.5 M.     

Auxin and cytokinin relationships in 24 microalgal strains1

Endogenous auxins and cytokinins were quantitated in 24 axenic microalgal strains from the Chlorophyceae, Trebouxiophyceae, Ulvophyceae, and Charophyceae. These strains were in an exponential growth phase, being harvested on day 4. Acutodesmus acuminatus Mosonmagyaróvár Algal Culture Collection‐41 (MACC) produced the highest biomass and Chlorococcum ellipsoideum MACC‐712 the lowest biomass. The auxins, indole‐3‐acetic acid (IAA) and indole‐3‐acetamide (IAM) were present in all microalgal strains. No other auxin conjugates were detected. IAA and IAM concentrations varied greatly, ranging from 0.50 to 71.49 nmol IAA · g^?1 DW and 0.18 to 99.83 nmol IAM · g^?1 DW, respectively. In 19 strains, IAA occurred in higher concentrations than IAM. Nineteen cytokinins were identified in the microalgal strains. Total cytokinin concentrations varied, ranging from 0.29 nmol · g^?1 DW in Klebsormidium flaccidum MACC‐692 to 21.40 nmol · g^?1 DW in Stigeoclonium nanum MACC‐790. The general trend was that cis‐zeatin types were the predominant cytokinins; isopentenyladenine‐type cytokinins were present in moderate concentrations, while low levels of trans‐zeatin‐type and very low levels of dihydrozeatin‐type cytokinins were detected. Ribotides were generally the main cytokinin conjugate forms present with the cytokinin free bases and ribosides present in similar but moderate levels. The levels of O‐glucosides were low. Only one N‐glucoside was detected, being present in nine strains in very low concentrations. In 15 strains, the auxin content was 2‐ to 4‐fold higher than the cytokinin content.     

Fatty Acid Composition of Pseudomonas syringae pv. savastanoi

Over 85% of total cellular fatty acids of 30 strains of P. syringae pv. savastanoi, grown for one day at 28 °C on King's medium B (KB) agar, were 12:0 (5.0%), 16:0 (27.5%), 16:1 (36.7%) and 18:1 (16.8%). Three hydroxy-substituted fatty acids comprised 7.2% of the total and 22 other minor components, each occurring at concentrations of less than 1%, comprised an additional 4%. Three percent were unidentified components. Cells grown for 3 and 6 days on KB agar contained lower concentrations of the unsaturated 16:1 (30.4 and 21.1%, respectively), and higher concentrations of branched-chain and cyclopropane fatty acids than one-day old cells. No consistent differences in fatty acid composition could be detected between virulent and avirulent strains, nor between pv. savastanoi and other pathovars of P. syringae. However, when cells were grown on a chemically-defined medium for 6 days, concentrations of 16:0 and a tentatively-identified 17-carbon hydroxy fatty acid were higher, and those of 12:0 and 16:1 were lower in strains from Fraxinus than from Olea. P. fluorescens (7 strains) and P. viridiflava (6 strains) could be differentiated from each other but not from P. syringae.     

Use of Trichoderma harzianum and Gliocladium vitens for the Biological Control of Post-emergence Damping-off and Root Rot of Cucumbers Caused by Pythium ultimum

The antagonist strains Gliocladium virens G2 and Trichoderma harzianum T3 originally isolated from Pythium suppressive peat, and two benomyl-resistant strains of T. harzianum, T12B and T95, were evaluated as biological control agents of damping-off and root rot of cucumbers in sphagnum peat caused by Pythium ultimum. All strains were equally effective when applied as 1 % peat-bran preparations, whereas the effectiveness of disease control was reduced at higher concentrations of the antagonists. The two wild-type strains were also found to be effective when applied as conidial suspensions, and in this case no reduction in disease control was seen at higher concentrations. G. virens G2 and T. harzianum T12B showed antibiotic activity against P. ultimum in in vitro tests; however there were no signs of mycoparasitism of P. ultimum by any of the antagonist strains.     

The effect of netilmicin, amikacin, ceftazidime and cefotaxime on the adhesive properties of microorganisms isolated from newborn infants]

The effect of netilmycin, amikacin, ceftazidime and cefotaxime on adhesion of Lactobacillus spp. (14 strains), Escherichia coli (21 strains), Klebsiella pneumonia (15 strains), Enterococcus sp. (18 strains), Candida albicans (15 strains) was investigated. The strains were isolated from respiratory tract and feces of the newborns. Antibiotics were used in the following subtherapeutic and therapeutic concentrations: netilmycin--1.2 and 12.0 micrograms/ml, amikacin--1.8 and 18 micrograms/ml, ceftazidime--7.5 and 75 micrograms/ml, cefotaxime--6.5 and 75 micrograms/ml. Adhesion of C. albicans was investigated with buccal epithelium cells, adhesion of other microorganisms--on formalinized human erythrocytes (1(0)Rh(+)). It was shown that antibiotics in subtherapeutic and therapeutic concentrations inhibited adhesion of the most strains. Cefalosporins demonstrated maximum inhibitory activity. The number of the strains inhibited by cefalosporins and by aminoglycosides enhanced along with antibiotics concentrations enhancement from subtherapeutic to therapeutic concentrations.     

Effect of N-nitroso-N-methylurea on viability and mutagenic response of repair-deficient strains of Escherichia coli

Various E. coli mutants, deficient in DNA repair, differed in their response to increasing concentrations of N-nitroso-N-methylurea (NMU).Loss of viability due to exposure to NMU was greatest in those strains with a reduced capacity for repair of single-strand breaks. Viability of wild-type and uvrA^? strains was not affected by NMU concentrations up to 3.0 mM. Some loss of viability occurred, at the higher NMU concentrations, in both strains carrying exrA^? while strains carrying uvrA^?polA^? or recA^? were the most sensitive. The results support the hypothesis that the lethal effect of NMU on repair-deficient E. coli was due to its ability to induce single-strand breaks.Induction of mutations by NMU was observed in all the strains used and the results suggested that NMU damage per se was the major mutational event. The dose response curve for induction of revertants by NMU was, however, influenced by the repair system(s) present. The number of revertants scored at the higher NMU concentrations was greater in those strains lacking the recA and polA dependent repair functions than in the wild-type strain. However, at NMU concentrations below 2.0 mM the numbers of revertants induced in exrA^? carrying strains, prossessing accurate rec-dependent repair, were lower than the comparable wild-type values. The evidence suggests that the uvrA gene product also acts on some, possibly non-mutagenic, types of NMU damage and that error-prone repair of these lesions increases the number of potential revertants.     

Bacteria--degraders of polycyclic aromatic hydrocarbons, isolated from soil and bottom sediments in salt-mining areas

Fifteen bacterial strains capable of utilizing naphthalene, phenanthrene, and biphenyl as the sole sources of carbon and energy were isolated from soils and bottom sediments contaminated with waste products generated by chemical and salt producing plants. Based on cultural, morphological, and chemotaxonomic characteristics, ten of these strains were identified as belonging to the genera Rhodococcus, Arthrobacter, Bacillus, and Pseudomonas. All ten strains were found to be halotolerant bacteria capable of growing in nutrient-rich media at NaCl concentrations of 1-1.5 M. With naphthalene as the sole source of carbon and energy, the strains could grow in a mineral medium with 1 M NaCl. Apart from being able to grow on naphthalene, six of the ten strains were able to grow on phenanthrene; three strains, on biphenyl; three strains, on octane; and one strain, on phenol. All of the strains were plasmid-bearing. The plasmids of the Pseudomonas sp. strains SN11, SN101, and G51 are conjugative, contain genes responsible for the degradation of naphthalene and salicylate, and are characterized by the same restriction fragment maps. The transconjugants that gained the plasmid from strain SN11 acquired the ability to grow at elevated NaCl concentrations. Microbial associations isolated from the same samples were able to grow at a NaCl concentration of 2.5 M.