Antifungal properties of some pyrazolyl-alkyl-sulfides

Four 5-amino-4-alkylthio-pyrazoles were synthesized and their antifungal activity was evaluated in vitro in Trichophyton mentagrophytes, Microsporum cookei and Candida albicans. The compounds slightly influenced the growth kinetics of the yeast, but at concentrations ranging from 20 to 40 g/ml completely prevented the mycelial growth of the two dermatophytes cultivated on Sabouraud's agar medium. An electron microscopic study, undertaken by using the most active compound, showed that in C. albicans mitochondria were the only cell targets affected whereas in the dermatophytes cell wall, plasmalemma and the main cytoplasmic organelles were damaged in various degrees. Since the most remarkable alterations were connected with membrane abnormalities, the cytological changes observed were tentatively interpreted as a consequence of the compound intrusion into the lipid bilayer of the membranes, since the drug is lipophilic in nature.Investigations supported by a grant from Consiglio Nazionale delle Ricerche of Italy (Contract No. 79.01871.04)     

Antifungal properties of cranberry juice

[This corrects the article on p. 1524 in vol. 16.].     

Speciation in the large-spored Metschnikowia clade and establishment of a new species, Metschnikowia borealis comb. nov

The reproductive boundaries among species in the large-spored Metschnikowia clade were studied by prototrophic recombinant selection, electrophoretic karyotyping, mitochondrial DNA restriction analysis, and DNA sequence analysis. Inviable ascospores arose from crosses between the two varieties of Metschnikowia continentalis, indicating that they should be recognized as separate species. Prototrophic recombinants were recovered from crosses between auxotrophic mutants of Metschnikowia borealis, M. continentalis, Metschnikowia lochheadii, Metschnikowia sp. UWO(PS)00-154.1, and Candida ipomoeae, showing that some genetic exchange is possible in spite of the sterility of the asci formed in interspecific crosses. Metschnikowia hawaiiensis, although capable of ascus formation when its h(-) mating type is crossed with the h(+) mating type of the other species, did not give rise to recombinants. In the other species, some recombinants acquired the ability to form asci directly from single cells. These often contained the chromosomes of both parents, suggesting formation of allodiploid hybrids. Other recombinants behaved as haploids and were similar to one parent except for having inherited the selectable wild-type allele from the other parent. In most, but not all cases, inheritance of the mitochondrial genome was uniparental and correlated with the inheritance of the nuclear chromosome complement. In some cases, what appeared to be a recombinant mitochondrial genome was observed. Phylogenies derived from the sequences of various DNA regions were not congruent, indicating that hybridization may have taken place in nature as the large-spored species diverged from their common ancestor. Further evidence that C. ipomoeae arose from a natural recombination event was obtained, but a pair of Metschnikowia species that might represent derived forms of the parents could not be identified conclusively. C. ipomoeae and most of its closely related Metschnikowia species contained a group-II intron in the mitochondrial small-subunit ribosomal gene. The intron was absent in M. borealis, M. hawaiiensis, and other species in the genus Metschnikowia.     

The effect of strain rate on the mechanical properties of human cortical bone

Bone mechanical properties are typically evaluated at relatively low strain rates. However, the strain rate related to traumatic failure is likely to be orders of magnitude higher and this higher strain rate is likely to affect the mechanical properties. Previous work reporting on the effect of strain rate on the mechanical properties of bone predominantly used nonhuman bone. In the work reported here, the effect of strain rate on the tensile and compressive properties of human bone was investigated. Human femoral cortical bone was tested longitudinally at strain rates ranging between 0.14-29.1 s(-1) in compression and 0.08-17 s(-1) in tension. Young's modulus generally increased, across this strain rate range, for both tension and compression. Strength and strain (at maximum load) increased slightly in compression and decreased (for strain rates beyond 1 s(-1)) in tension. Stress and strain at yield decreased (for strain rates beyond 1 s(-1)) for both tension and compression. In general, there seemed to be a relatively simple linear relationship between yield properties and strain rate, but the relationships between postyield properties and strain rate were more complicated and indicated that strain rate has a stronger effect on postyield deformation than on initiation of yielding. The behavior seen in compression is broadly in agreement with past literature, while the behavior observed in tension may be explained by a ductile to brittle transition of bone at moderate to high strain rates.     

DNA relatedness among aquatic yeasts of the genus Metschnikowia and proposal of the species Metschnikowia australis comb. nov

DNA hybridization studies were conducted to determine the taxonomic status of the aquatic group of Metschnikowia species and their varieties. Among the DNAs of the four varieties of Metschnikowia bicuspidata, that of Metschnikowia bicuspidata var. australis showed 37 to 51% relative binding with the DNAs of Metschnikowia bicuspidata var. bicuspidata and of the varieties chathamia and californica. On this basis, low intervarietal fertility, and unique habitat in antarctic seawater, we have proposed to raise M. bicuspidata var. australis to the rank of species, Metschnikowia australis, comb. nov. DNA complementarity of Metschnikowia bicuspidata var. californica and Metschnikowia bicuspidata var. chathamia DNA was greater than 80% with that of M. bicuspidata var. bicuspidata. Metschnikowia australis can be differentiated from other Metschnikowia species and varieties by its inability to form chlamydospores, the formation of two needle-shaped ascospores per ascus, lack of glucose fermentation, and lack of assimilation of both methyl-alpha-D-glucoside and glucono-delta-lactone. Other DNA-DNA reassociation experiments showed that Metschnikowia zobellii is a distinct species when compared with both aquatic and terrestrial species of the genus.     

Antifungal properties of new series of quinoline derivatives

The series of quinoline derivatives were prepared. The synthetic approach, analytical, and spectroscopic data of all synthesized compounds are presented. All the prepared derivatives were analyzed using the reversed-phase high performance liquid chromatography (RP-HPLC) method for the lipophilicity measurement. In the present study, the correlation between RP-HPLC retention parameter log K (the logarithm of capacity factor K) and various calculated log P data is shown. The relationships between the lipophilicity and the chemical structure of the studied compounds are discussed as well. The prepared compounds were tested for their in vitro antifungal activity. 2-[(3-Hydroxyphenylimino)methyl]quinolin-8-ol (8), 2-[(4-hydroxyphenylimino)methyl]quinolin-8-ol (9) and 2-[(2,5-dichloro-4-nitrophenylamino)methoxymethyl]quinolin-8-ol (10) showed in vitro antifungal activity comparable to or higher than that of the standard fluconazole. Structure-activity relationships among the chemical structure, the physical properties, and the biological activities of the evaluated compounds are discussed in the article.     

Antifungal properties of haem peroxidase from Acorus calamus

BACKGROUND AND AIMS: Plants have evolved a number of inducible defence mechanisms against pathogen attack, including synthesis of pathogenesis-related proteins. The aim of the study was to purify and characterize antifungal protein from leaves of Acorus calamus. METHODS: Leaf proteins from A. calamus were fractionated by cation exchange chromatography and gel filtration and the fraction inhibiting the hyphal extension of phytopathogens was characterized. The temperature stability and pH optima of the protein were determined and its presence was localized in the leaf tissues. KEY RESULTS: The purified protein was identified as a class III haem peroxidase with a molecular weight of approx. 32 kDa and pI of 7.93. The temperature stability of the enzyme was observed from 5 degrees C to 60 degrees C with a temperature optimum of 36 degrees C. Maximum enzyme activity was registered at pH 5.5. The pH and temperature optima were corroborated with the antifungal activity of the enzyme. The enzyme was localized in the leaf epidermal cells and lumen tissues of xylem, characteristic of class III peroxidases. The toxic nature of the enzyme which inhibited hyphal growth was demonstrated against phytopathogens such as Macrophomina phaseolina, Fusarium moniliforme and Trichosporium vesiculosum. Microscopic observations revealed distortion in the hyphal structure with stunted growth, increased volume and extensive hyphal branching. CONCLUSIONS: This study indicates that peroxidases may have a role to play in host defence by inhibiting the hyphal extension of invading pathogens.     

Antifungal properties of selected plants from the Yucatan peninsula,Mexico

A total of 20 plants belonging to different genera (Acalypha, Ageratum, Ambrosia, Bidens, Blechum, Caesalpinia, Calea, Carlowrightia, Croton, Eugenia (2), Furcraea, Stenandrium, Tephrosia, Trichilia (2), Randia (3), and Vitex) were selected from native flora of the Yucatan peninsula. All plants selected were collected and separated in to leaves, stems and roots. These were then extracted with ethanol and their crude extracts (54) were evaluated against Alternaria tagetica, Colletotrichum gloeosporioides, Fusarium oxysporum and Rhizopus sp. using the filter paper disc diffusion assay. Results lead to the selection of 33 crude extracts active against at least one of the target strains, which were assessed to determine their ability to inhibit the mycelial growth of the pathogenic fungi in a second antifungal assay. The results of this assay indicated that extracts from the roots of Croton chichenensis were the most promising, with a wide activity spectrum against all pathogens tested in both assays with inhibition percentages of greater than 60%. Furthermore, extracts from leaves of Ambrosia hispida, Trichilia minutiflora, and roots of Acalypha gaumeri were able to cause growth inhibition against two or three pathogen strains (≥50%). Studies of these active extracts should be continued at different levels. In general, results revealed a good bioactive potential of the flora from the Yucatan peninsula to produce metabolites with potential applications as botanical pesticides in the near future.     

Antifungal characteristics of a fluorescent Pseudomonas strain involved in the biological control of Rhizoctonia solani

A plant growth-promoting isolate of a fluorescent Pseudomonas spp. EM85 was found strongly antagonistic to Rhizoctonia solani, a causal agent of damping-off of cotton. The isolate produced HCN (HCN+), siderophore (Sid+), fluorescent pigments (Flu+) and antifungal antibiotics (Afa+). Tn5::lacZ mutagenesis of isolate EM85 resulted in the production of a series of mutants with altered production of HCN, siderophore, fluorescent pigments and antifungal antibiotics. Characterisation of these mutants revealed that the fluorescent pigment produced in PDA and the siderophore produced in CAS agar were not the same. Afa- and Flu- mutants had a smaller inhibition zone when grown with Rhizoctonia solani than the EM85 wild type. Sid- and HCN mutants failed to inhibit the pathogen in vitro. In a pot experiment, mutants deficient in HCN and siderophore production could suppress the damping-off disease by 52%. However, mutants deficient in fluorescent pigments and antifungal antibiotics failed to reduce the disease severity. Treatments with mutants that produced enhanced amounts of fluorescent pigments and antibiotics compared with EM85 wild type, exhibited an increase in biocontrol efficiency. Monitoring of the mutants in the rhizosphere using the lacZ marker showed identical proliferation of mutants and wild type. Purified antifungal compounds (fluorescent pigment and antibiotic) also inhibited the fungus appreciably in a TLC bioassay. Thus, the results indicate that fluorescent pigment and antifungal antibiotic of the fluorescent Pseudomonas spp. EM85 might be involved in the biological suppression of Rhizoctonia-induced damping-off of cotton.     

Synonymy of Metschnikowia pulcherrima and Torulopsis burgeffiana

Torulopsis burgeffiana is placed in the genus Metschnikowia because it has been shown to have a perfect stage corresponding with this genus. Due to the characteristic shape of the ascus and ascospores, T. burgeffiana is to be considered a synonym of M. pulcherrima. The physiological differences which exist between strains are not sufficiently important to maintain a separate species.     

Production of carbonyl reductase by Metschnikowia koreensis

A new strain of the yeast Metschnikowia koreensis was grown in shake flasks and a stirred bioreactor for the production of carbonyl reductase. The optimal conditions in the bioreactor for maximizing the biomass specific activity of the enzyme were found to be: a medium composed of glucose (20 g/L), peptone (5 g/L), yeast extract (5 g/L) and zinc sulfate (0.3g/L); the pH controlled at 7; the temperature controlled at 25 °C; an agitation speed of 500 rpm; and an aeration rate of 0.25 vvm. In the bioreactor, a biomass specific enzyme activity of 115.6 U/gDCW was obtained and the maximum biomass concentration was 15.3 gDCW/L. The biomass specific enzyme activity obtained in the optimized bioreactor culture was 11-fold higher than the best result achieved in shake flasks. The bioreactor culture afforded a 2.7-fold higher biomass concentration than could be attained in shake flasks.     

Antifungal properties of Singapore gorgonians: a preliminary study

Gorgonians possess a huge array of secondary metabolites for various functions, many of which are not known. One of these functions is antifungal. This study investigates if gorgonians from reefs in Singapore can defend themselves against the settlement and invasion of fungi. Crude extracts from 10 species of gorgonians from three families, Ellisellidae, Subergorgiidae and Plexauridae, were screened against nine species of deuteromycetous fungi previously isolated from gorgonians. Minimum Inhibitory Concentration (MIC) experiments were carried out in 96-microwell plates using extract concentrations ranging from 1.5 to 24.0 mg/ml. Extracts from Euplexaura cf. pinnata, Echinogorgia sp. C, Junceella cf. gemmacea, Subergorgia suberosa, Ctenocella cf. umbraculum and Junceella sp. A were found to possess inhibitory effects on fungi. MICs range from 1.5 to 18.0 mg/ml. Results showed that most of the antifungal activities were exhibited by Euplexaura cf. pinnata and Echinogorgia sp. C from the family Plexauridae. However, for most gorgonian species, the concentrations required to inhibit fungal growth are much higher than the natural concentrations of extracts found in gorgonian tissues. Only the extracts of Echinogorgia sp. C, C. cf. umbraculum and S. suberosa were found to inhibit fungal growth of a few fungal species at a concentration lower than that of its natural concentration.     

Infection of marine copepods by Metschnikowia sp.

Summary It was observed that yeasts belonging to the genusMetschnikowia parasitize marine copepods both under natural and experimental conditions. The physiological activity ofMetschnikowia sp. was investigated under a variety of conditions and the importance of these organisms in the ecology of the Strait of Georgia has been discussed.