Human dermatophytosis caused by Trichophyton equinum

A case of dermatophytosis caused by Trichophyton equinum is reported. A 25-year-old man employed at a breeding center of a horse racing course was infected on the left arm in August, 1981. The lesion had a vesicle or a small pustule accompanied by severe itching. The fungal elements of the scale were identified by microscopic observation. Griseofulvin administration was found to be very effective for treating this infection. In a mycological examination, T. equinum was isolated mainly on cycloheximide-chloramphenicol Sabouraud's dextrose agar and chloramphenicol potato dextrose agar. Equine dermatophytosis was quite prevalent at this race course, so that this area as well as equipment used for the maintenance and care of the horse was very likely to be the source of the patient's infection.     

A new approach of pellet formation of a filamentous fungus -Rhizopus oryzae

The effects of inoculum and medium composition (i.e. potato dextrose broth as carbon source, soybean peptone, calcium carbonate, and metal ions) on pellet formation of Rhizopus oryzae ATCC 20344 have been studied. Metal ions were found to have a significant negative effect on pellet formation while soybean peptone had a positive effect. In addition, potato dextrose broth and calcium carbonate were beneficial to R. oryzae for growing small, smooth pellets during the culture. The study also demonstrated that an inoculum size of less than 1.5x10(9)spores/L had no significant influence on pellet formation although it had large impacts on pellet growth. Thus, a new approach to form pellets has been developed using only potato dextrose broth, soybean peptone, and calcium carbonate allowing for pellet size to be controlled by adjusting inoculum size and the concentrations of potato dextrose broth, soybean peptone, and calcium carbonate in the medium.     

Identification of bacterial species on Lippia multiflora herbal tea leaves and the influence of steam pasteurization

The consumption of herbal teas is an increasing phenomenon among tea consumers globally. Some of these herbal teas are not pre-treated to reduce their microbial load before consumption, and thus constitute a health risk to consumers. In this study, the effect of steam pasteurization, at >99 °C for 2.5 min, on the microbial load of Lippia multiflora herbal tea leaves was evaluated. Microbial enumeration was conducted on potato dextrose agar, plate count agar, violet red bile agar, yeast peptone dextrose agar, and DeMan-Rogosa-Sharpe agar. Morphologically distinct colonies were isolated, sub-cultured and their Gram reaction recorded. These bacteria were identified to the species level using 16S ribosomal DNA sequence data. Most of the bacteria identified belonged to the genus Bacillus. One species each from the genera Pantoea and Kocuria was also identified, but only the Bacillus species survived the steam pasteurization treatment. Coliform bacteria detected prior to pasteurization were not detected after the steam treatment. Steam pasteurization reduced the microbial load from 10⁴ to 10² c.f.u.g⁻¹and it is potentially an effective method to treat L. multiflora herbal teas prior to consumption. It is important to note that the steam treatment should complement good agricultural and hygienic practices rather than replace them, as some bacteria can survive this treatment.     

Screening for phenoloxidases from edible mushrooms.

A screening test for phenoloxidases from edible mushrooms was done on potato dextrose agar plates that contained phenolic chemicals. Many edible mushrooms showed positive reactions on the agar plates. Among them, Auricularia auricula-judae, Clitocybe nebularis, Lentinus edodes, Pholiota aurivella, and Pseudohiatula oshimae produced a considerable amount of phenoloxidases, and these enzymes showed maximum activities in the acidic pH region.     

COMPARISONS OF DIFFERENT MEDIA FOR COUNTING SUGAR TOLERANT YEASTS IN CONCENTRATED ORANGE JUICE

SUMMARY: To select and count the sugar tolerant yeasts which ferment sixfold concentrated orange juice, a high sugar agar medium was developed which contains 50% of glucose, 1% of citric acid and 1% of Tryptone; it is incubated for 4–5 days at 25°.
The medium has disadvantages: it is troublesome to prepare, and colonies grow slowly and are translucent. These properties result directly' from the high sugar concentration, on which the selective action of the medium depends.
Counts on this medium have been compared with those on potato dextrose or nutrient dextrose agars (with 2% and 1% of glucose respectively), with yeasts isolated from fermenting concentrate, in pure culture, and under various practical conditions. As a rule, the counts were virtually the same on the different media; nutrient dextrose agar occasionally failed to record small numbers of these yeasts. If the two low sugar media were acidified to pH 3·5 the counts were reduced.
Potato dextrose agar recorded, besides the above yeasts, sugar intolerant yeasts entering from dirty machines or through bad canning practice: nutrient dextrose agar recorded bacteria in addition. The difference between parallel plating on these media and on the high sugar medium thus yielded useful information about sources of casual contamination.
It is suggested that the above would also be largely applicable to other sugar-rich concentrates of not less than 50° Brix.     

Ethanol-sensitive mutants of Saccharomyces cerevisiae

Saccharomyces cerevisiae mutants unable to grow at ethanol concentrations at which the wild type strain S288C does grow, have been isolated. Some of them show additional phenotypic alterations in colony size, temperature sensitivity and viability in ethanol, which cosegregate with the growth sensitivity in ethanol. 21 selected monogenic ethanol-sensitive mutants define 20 complementation groups, denominated ETA1 to ETA20, which indicates that there is a high number of genes involved in the ethanol tolerance/sensitivity mechanism.Out of 21 selected monogenic mutants, 20 are not altered in the glycolytic pathway since, when maintained in glucosesupplemented medium, they can produce as much ethanol as the wild type and at about the same velocity. Nor do any of the mutants seem to be altered in the lipid biosynthetic pathway since, whether grown in the absence or in the presence of ethanol, their concentration of fatty acids and ergosterol is similar to that of the wild type under the same conditions. Therefore growth sensitivity to ethanol does not seem necessarily to be related to carbohydrate or lipid metabolism.Non-common abbreviations YP yeast extract peptone medium - YPD yeast extract peptone dextrose agar or medium - YPG yeast extract peptone glycerol agar - YPDE yeast extract peptone dextrose ethanol agar or medium - SD yeast nitrogen base dextrose agar - SPO yeast extract potassium acetate glucose agar - PD parental ditype - NPD non-parental ditype - TT tetratype     

Influence of <Emphasis Type="Italic">Flammulina velutipes</Emphasis> mycelia culture conditions on antimicrobial metabolite production

Enokipodins A, B, C, and D are α-cuparene-type sesquiterpenoids antimicrobial metabolites produced in the stationary stage of Flammulina velutipes mycelia development in malt extract broth. This study assessed the influence of nutritional and environmental factors on F. velutipes mycelia culture for the production of these metabolites. The mycelia growth and antimicrobial activity were assessed by determining dry matter and the diffusion in agar method, respectively. The best F. velutipes mycelia growth was observed in dextrose potato broth, and greater antimicrobial metabolite production occurred in complete Pontecorvo’s culture medium. Environmental modifications, such as a rise in temperature from 25° to 37°C on the 15th day of F. velutipes mycelia culture in malt extract and peptone broth, also optimized antimicrobial metabolite production. The metabolites produced in these treatments were correlated with the enokipodins A and B in thin-layer chromatography (TLC) and the antifungal activity test by TLC bioautography. This study showed that there was no correlation between biomass production and antimicrobial metabolite production, but there may be a correlation between culture medium composition and enokipodins biosynthesis.     

Screening for Phenoloxidases from Edible Mushrooms

A screening test for phenoloxidases from edible mushrooms was done on potato dextrose agar plates that contained phenolic chemicals. Many edible mushrooms showed positive reactions on the agar plates. Among them, Auricularia auricula-judae, Clitocybe nebularis, Lentinus edodes, Pholiota aurivella, and Pseudohiatula oshimae produced a considerable amount of phenoloxidases, and these enzymes showed maximum activities in the acidic pH region.     

In vitro propagation and mass scale multiplication of a critically endangered epiphytic orchid, Gastrochilus calceolaris (Buch.-Ham ex J.E.Sm.) D.Don. using immature seeds

In vitro asymbiotic seed germination potential of its immature seeds (36 weeks after pollination) of G. calceolaris was successfully tested on three different agar gelled nutrient media i.e. Murashige and Skoog (MS), Mitra et al. (M) and potato dextrose agar (PDA). Seeds germinated within 15.75+/-0.75 to 35.75+/-0.75 days in the three different media. The protocorms developed therefrom subsequently differentiated into first leaf and root primordia, and complete seedlings were obtained within 111.25+/-1.25 to 141.25+/-1.25 days on MS and M media. The protocorms, though failed to differentiate further on basal PDA medium, despite repeated subculturings, incorporation of peptone (P; 1 gl(-1)), yeast extract (YE; 2 gl(-1)) and coconut water (CW; 20%) in the medium proved beneficial in inducing differentiation, in these germinating entities. Additional use of growth additives (P/YE/CW), in general, favoured better germination, protocorm formation and seedling development. The optimal nutritional combination during seed germination, protocorm growth and multiplication and seedling development was found to be CW (10%) enriched MS medium.     

Production of Botrytis cinerea for potential introduction into a vineyard

Botrytis cinerea was produced in solid-phase fermentation, liquid fermentation and on potato dextrose agar. Stored products were evaluated for grape colonization in grape bioassays and in field trials, and for B. cinerea density using colony forming unit analyses and a nucleic-acid-based method. B. cinerea colony forming unit density was significantly correlated to the probability of successful grape colonization in grape bioassays (p-value=0.0002). Solid fermentation products could be stored longer than liquid fermentation and potato dextrose agar products. There was little difference in the rate of grape colonization in laboratory bioassays among solid-phase fermentation, liquid fermentation and plate culture products. Although the initial B. cinerea colonization rate of field grapes was slightly greater on vines treated with solid-phase fermentation and plate culture products compared to vines treated with product from liquid fermentation, there was no significant difference in final colonization between vines treated with solid-phase fermentation, liquid fermentation and plate culture products and untreated vines.     

Culture medium for selective isolation and enumeration of Gram-negative bacteria from ground meats.

We developed a new medium, designated peptone bile amphotericin cycloheximide (PBAC) agar, which contains (per liter) 10 g of peptone, 300 mg of bile salts, 1 mg of amphotericin B, 1 g of cycloheximide, and 15 g of agar. When 21 samples of fresh ground beef were studied and plate count agar counts were used as references, we obtained a mean recovery of 28% of total counts with violet red bile agar overlay, whereas we obtained 48% recovery with PBAC agar. With 12 samples of frozen ground beef, recovery on violet red bile agar overlay was 29% of the recovery on plate count agar, whereas the corresponding value on PBAC agar was 45%. PBAC agar allowed the enumeration of 1.4 times as many gram-negative bacteria as violet red bile agar overlay. None of eight strains of gram-positive bacteria and none of eight strains of yeasts grew on PBAC agar. Of 158 colonies randomly selected from pour plates of eight fresh ground meat samples, 95% stained gram negative. In comparison, only 70% of 151 colonies selected from corresponding plate count agar plates were gram negative. The lack of background color, turbidity, and ease of use make PBAC agar easier to handle than other media used for gram-negative bacteria, such as violet red bile agar, violet red bile agar overlay, and crystal violet tetrazolium agar. In the preparation PBAC agar, all ingredients are autoclaved together except amphotericin B, which is filter sterilized and added before the plates are poured.     

Transfer of a benomyl resistance marker by heat-inactivated Trichoderma reesei protoplasts

Summary Protoplasts from a benomyl resistant Trichoderma reesei mutant were heat inactivated at 60°C for 8 min and fused with viable protoplasts from an osmosensitive, non-sporulating T. reesei strain. Fusants recovered on 50 g/ml benomyl containing potato dextrose agar plates grew and sporulated well. Cellulolytic enzyme activities produced in liquid culture by selected fusants were higher than those produced by parental strains.     

Effect of supplementall-tyrosine on pigment production in cultures of the Legionnaires' disease bacterium

The etiologic agent of Legionnaires' disease grows on certain agar media. Cultures of this organism on supplemented Mueller-Hinton agar are characterized by the appearance of brown pigment in and around areas of bacterial growth. The major peptone source in Mueller-Hinton agar is an acid hydrolysate of casein. Legionnaires' disease bacterium also grows on a medium in which the peptone source is 0.25% yeast extract, but no pigment is produced. If the yeast extract agar is enriched withl-tyrosine, pigment formation can occur. Pigmentation of cultures of Legionnaires' disease bacterium may be mediated by a phenolo-monooxygenase, or tyrosinase.     

Production of antibiotic substances by natural variants of the marine bacterium Vibrio Fischeri

It was shown that under definite conditions there was competition between natural variants of sea bacteria belonging to V. fischeri. Natural variants of V. fischeri, strain 6 differed in their resistance to streptomycin and had different growth rates under conditions of limited aeration. Morphologically all the variants were identical. V. fischeri P-0, V. fischeri P-1 and V. fischeri P-2 were studied. The study revealed that V. fischeri P-0 produced a non-dialysing thermostable trypsin-sensitive substance inhibiting the growth of V. fischeri P-1 and V. fischeri P-2. The maximum activity of the antibacterial substance was observed when V. fischeri P-0 was grown in a liquid medium with peptone and yeast extract without agitation at 26 degrees C. The inhibiting substance was also active against V. fischeri BKM B995 and V. fischeri P-7 isolated as a result of V. fischeri P-0 exposure to ethidium bromide. The substance had no effect on the following bacterial species: Aeromonas liquefaciens 301, Achromobacter liquefaciens, Pseudomonas putida 15, Pseudomonas fluorescence 7, Escherichia coli AH-32 and Staphylococcus aureus.     

Biological and physiological characteristics of Neotyphodium gansuense symbiotic with Achnatherum inebrians

Biological and physiological characteristics of Neotyphodium gansuense were compared with Neotyphodium coenophialum and Epichlo? festucae at a range of temperatures and pH values, and on carbon and nitrogen amended media. N. gansuense was able to grow at 10-30 degrees C, but not at 5 degrees C, and slowly at 35 degrees C. The optimal temperature for both N. gansuense and N. coenophialum was 25 degrees C, but that of E. festucae was 20-25 degrees C. The optimal pH ranges for mycelial growth of N. gansuense, N. coenophialum and E. festucae were 5-9, 5-9 and 5-7, respectively. The Neotyphodium and Epichlo? endophytes varied in their ability to grow on media containing different carbon and nitrogen nutrients. The preference of N. gansuense for carbon source was sucrose>glucose, lactose, sorbitol, inulin, maltose, mannitol, starch, fructose>xylose. Growth of all three endophytes tested was significantly improved by peptone, tryptone, casein, yeast extract and l-proline. Yeast extract, peptone, casein, tryptone, l-proline, potassium nitrate, ammonium oxalic acid and l-leucine significantly improved growth of N. gansuense. However, ammonium nitrite was not utilized at all by any tested endophyte. N. gansuense grew significantly better on potato dextrose agar (PDA) and oat meal agar (OMA) than on corn meal agar (CMA) and drunken-horse-grass agar (DA), and most slowly on water agar (WA) and saltwater nutrient agar (SNA).     

Effects of the demethylase inhibitor,cyproconazole, on hyphal tip cells ofSclerotium rolfsii: II. An electron microscope study

Hyphal tip cells ofSclerotium rolfsii, prepared by freeze substitution, were examined by transmission electron microscopy after continued growth on cyproconazole-amended potato dextrose agar at fungistatic concentrations of 0.1, 0.75, and 1.0 μg/ml which respectively produced 50, 80, and 95% growth inhibition. Alternatively, observations were made of control tip cells that were transferred onto potato dextrose agar amended with 0.75 μg/ml cyproconazole for 30, 60, and 240 min. Ultrastructural abnormalities observed in hyphal tip cells as a result of cyproconazole treatment included (1) Spitzenko¨rper disorganization, (2) proliferation of smooth endoplasmic reticulum with inflated cisternae, (3) increased vacuolar contents, (4) presence of monovesicular bodies, and (5) irregular cell wall characteristics. Alterations of the Spitzenko¨rper, endoplasmic reticulum, and vacuoles were noted 30 min after exposing control hyphae to cyproconazole (0.75 μg/ml). Abnormal wall depositions were observed 60 min after fungicide exposure. After a 140-min exposure to fungicide, tip cell morphology was similar to that of cells grown in the continual presence of 0.75 μg/ml cyproconazole. The changes in hyphal morphology are discussed relative to the mode of action of cyproconazole.     

Permanent mounts of fungus cultures grown on media optimal for production of characteristic structures

Summary 1. Dermatophyte cultures in plastic flasks were compared on four different media for total growth, sporulation, and pigment production. Czapek's agar favored more rapid and more abundant sporulation in most cultures than Sabouraud's dextrose agar, potato dextrose agar, or wort agar.2. A simple technique for embedding agar island cultures of fungi is described. These plastic-embedded cultures form permanent mounts useful for study of microscopic morphology.3. A relatively durable slant culture in a similar plastic flask is suggested as a companion for study of gross morphologic features of the cultures.