Nosocomial Invasive Infection Caused by Cunninghamella bertholletiae: Case Report

Human infection by Cunninghamella bertholletiae occurs almost exclusively in immunocompromised patients. Infections due to this microorganism have been most frequently diagnosed in patients with hematological malignancies, with neutropenia and in diabetes mellitus patients. This work reports a case of fungal infection by Cunninghamella bertholletiae isolated from blood in a man with a complex clinical picture, involving diabetes and pharmacological immunosupression. Blood culture at room temperature and at 37 °C on Sabouraud agar grew a single mold with characteristic properties of Cunninghamella. In the microscopic morphology, were found wide, non-septate, branching hyphae with erect sporangiophores terminated in swollen vesicles and sporangioles borne off the vesicles. C. bertholetiae was identified after subculture on Sabouraud dextrose agar at 45 °C. The patient died 15 days after the beginning of amphotericin B therapy.     

Metabolism of cinnoline to N-oxidation products by Cunninghamella elegans and Aspergillus niger

Cultures of the fungi Cunninghamella elegans and Aspergillus niger were grown in fluid Sabouraud medium at 28°C for 3 days and then dosed with cinnoline (1,2-diazanaphthalene). After 3 more days, metabolites were extracted from the cultures with ethyl acetate, separated by high-performance liquid chromatography, and identified by mass spectrometry and proton nuclear magnetic resonance spectroscopy. Both fungi oxidized 2–10% of the added cinnoline to mixtures of cinnoline 2-oxide and cinnoline 1-oxide. Received 10 June 1998/ Accepted in revised form 25 September 1998     

An evaluation of straw-extract agar media for the growth and sporulation of Madurella mycetomatis

Four new media, namely Wheat straw extract agar, Bajra straw extract agar, Jowar straw extract agar and Paddy straw extract agar, were evaluated for their potential to stimulate the growth and sporulation of Madurella mycetomatis in comparison with the conventional Sabouraud dextrose agar and Soil extract agar. Vegetative growth of M. mycetomatis on the four types of Straw extract agars was superior to that obtained on Sabouraud dextrose agar. Isolates of M. mycetomatis sporulated better and faster on the Straw extract agars than on the Sabouraud dextrose agar and Soil extract agar. Straw extract agar is recommended as a sporulation medium for M. mycetomatis. It may prove useful especially for studies of the conidium ontogeny of the fungus for elucidating its taxonomic status.     

A DL-DOPA drop test for the identification of Cryptococcus neoformans

A simple melanin assay using DL.DOPA as the substrate was developed to aid in the identification of Cryptococcus neoformans. The DL-DOPA drop test assay was simple and efficient. The best results (100% of the C. neoformans isolates were positive) occurred when C. neoformans was grown for two days at room temperature on Sabouraud agar modified. One to three loopfuls of yeast cells were then transferred to a starvation medium for 18–24 hours. Two to three drops of 0.3% DL-DOPA solution was applied to the transferred yeast cells. Only C. neoformans produced a brown or blackgrey pigment within 24 hrs, with 85% of the isolates becoming brown or black-grey within thirty minutes.     

In vivo assessment of the effect of an adhesive for complete dentures on colonisation of Candida species

doi: 10.1111/j.1741‐2358.2009.00345.x
In vivo assessment of the effect of an adhesive for complete dentures on colonisation of Candida species Objective: Denture adhesives have long been recognised by denture wearers as a useful adjunct to denture retention and stability. The objective of the present study was to evaluate, in vivo, the effect of a denture adhesive on oral quantities of Candida species by determination of absolute counts of colony‐forming units (CFU) per ml of saliva of individuals who use this denture adhesive for a period of 14 days. Materials and methods: Twenty‐four individuals were randomised in two equal groups of 12 (test and control), with the individuals of the test group using the adhesive for 14 days. Samples of saliva were collected from all individuals on days 0 (initial), 7 and 14. Aliquots of saliva were diluted and plated in duplicate on Sabouraud dextrose agar with chloramphenicol and incubated for 37°C for 48 h, the CFU/ml were counted in the individuals of each group and the data of each group were compared at the different time periods and analysed statistically by the non‐parametric Mann‐Whitney U‐test (α ≤ 5%). Results: There were no statistically significant differences between the test and control groups during the test periods. Conclusion: Within of the limitations of this study, the data suggested that the denture adhesive tested did not significantly alter the oral microbiota during the 14‐day trial period.     

Inhibition ofGeotrichum candidum by bicarbonate

Summary In a Sabouraud liquid medium system with NaHCO₃ or KHCO₃ at as low as 0.1% (w/v), the mycelial growth ofGeotrichum candidum was inhibited by greater than 90%. Bicarbonate may have a potential use as a microbial inhibitor in a variety of food systems.

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Inhibition de Geotrichum candidum par le bicarbonate

Résumé La croissance mycélienne deGeotrichum candidum a été inhibée à plus de 90% dans le milieu liquide de Sabouraud contenant le NaHCO₃ ou le KHCO₃ à des concentrations aussi faibles que 0.1%. Le bicarbonate peut ainsi avoir un usage potentiel comme inhibiteur microbien dans divers systèmes nutritifs.

     

Occurrence ofFusarium sacchari var.subglutinans and its mycotoxin production ability in broiler feed

Spores ofFusarium sacchari var.subglutinans isolated from broiler feed BR1 were obtained at an average concentration of 1.5/mg in 25% of tested samples. The spore concentration was increased from 1 to 100/mg of solid substrate (BR2; biscuit) or to 1/nL of Sabouraud broth after 3 weeks of cultivation. Mycotoxin analyses of these three substrates showed negative reactions for T-2 toxin and zearalenone but a positive reaction for deoxynivalenol (DON) which was found in concentrations of 5 ppm in Sabouraud broth, 50 ppm in BR2 and 220 ppm in biscuit. Therefore, ourF. sacchari isolate appeared to be a DON producer.     

First report of oral colonization by <Emphasis Type="Italic">Debaryomyces nepalensis</Emphasis> in a dog

A stray, young male, wire-haired pointing griffon dog, found in a street of Perugia (Italy), was examined in order to check his health status. Two oropharyngeal swabs were collected in 24 h and streaked onto Sabouraud agar and after 6 days the yeasts colonies were transferred onto Malt agar. Ascospores were observed on Potato Dextrose Agar medium. The major ubiquinone of an isolated yeast was identified as ubiquinone-9 (Q-9), and genetical analyses were performed together with the type strains of Debaryomyces hansenii (var. hansenii and var. fabry), C. psychrophila and D. nepalensis type strain. The base sequences of ITS1 and ITS2, and D1/D2 domains of LSU rDNA completely coincided with those of D. nepalensis. From these results, the isolated yeast was identified as D. nepalensis. RAPD patterns between the two strains were found to be identical. The results indicate the first colonization of D. nepalensis in a dog.     

Metabolism of the ethanolamine-type antihistamine diphenhydramine (Benadryl)TM by the fungus Cunninghamella elegans

Two strains of the filamentous fungus Cunninghamella elegans (ATCC 9245 and ATCC 36112) were grown in Sabouraud dextrose broth and screened for the ability to metabolize the ethanolamine-type antihistamine diphenhydramine. Based on the amount of parent drug recovered after 7 days incubation, both C. elegans strains metabolized approximately 74% of the diphenhydramine, 58% of this being identified as organic extractable metabolites. The organic extractable metabolites were isolated by reversed-phase high-performance liquid chromatography and identified by analyzing their mass and nuclear magnetic resonance spectra. Desorption chemical ionization mass spectrometry (DCIMS) with deuterated ammonia was used to differentiate possible isobaric diphenhydramine metabolites and to probe the mechanisms of ion formation under ammonia DCIMS conditions. C. elegans transformed diphenhydramine by demethylation, oxidation, and N-acetylation. The major metabolites observed were diphenhydramine-N-oxide (3%), N-desmethyldiphenhydramine (30%), N-acetyldidesmethyldiphenhydramine (13%), and N-acetyl-N-desmethyldiphenhydramine (12%). These compounds are known mammalian metabolites of diphenhydramine and may be useful for further toxicological studies. Received: 24 June 1999 / Revision received: 23 August 1999 / Accepted: 24 September 1999     

Effects of Temperature and Culture Media on Vegetative Growth of an Entomopathogenic Fungus <Emphasis Type="Italic">Isaria</Emphasis> sp. (Hypocreales: Clavicipitaceae) Naturally Affecting the Whitefly, <Emphasis Type="Italic">Bemisia tabaci</Emphasis> in Texas

The effects of temperature and mycological media on mycelial growth and estimates of spore production of an indigenous entomopathogenic fungus, Isaria sp., found during natural epizootics on whiteflies in the Lower Rio Grande Valley of Texas, were investigated. The radial growth (mm/day) of Isaria sp. as a function of temperature fits a linear model; with faster growth on Sabouraud dextrose agar with yeast extract, SDAY slopes (0.23) than on Sabouraud maltose agar, SMA slopes (0.14) from 20 to 30°C, with an optimal temperature of 30°C (SDAY: 4.1 mm, SMA: 3.1 mm). Moderate growth occurred at 25°C (SDAY: 3.4 mm, SMA: 2.7 mm). Growth was lowest at 20°C (SDAY: 1.9 mm, SMA: 1.8 mm). No fungal growth was observed at 35°C and 40°C. However, when Isaria sp. was exposed to 35°C for the first 7 days, it could recover and grow when transferred to 25°C (SDAY: 3.5 mm, SMA: 2.8 mm). No recovery or growth occurred after transfer from 40°C to 25°C. The average conidial production on SDAY after 20 days incubation at 25°C and a photoperiod of 14:10 h light: dark was 1.2 × 10⁸ conidia/cm² with 100% spore viability. When compared on SDAY at 25°C, the radial growth rate of I. javanica ex type CBS 134.22 (5.1 mm/day) was greater than seven Isaria isolates including Isaria sp.; but maximum growth rates were similar among all related Isaria isolates (90–97%). The Isaria sp. fungus tolerates high temperatures (35°C), suggesting that it is naturally selected for the subtropical semi-arid environment, where it could serve as an important natural control agent of the sweet potato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) biotype B, one of the most invasive and economically damaging insects to agriculture. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U. S. Department of Agriculture.     

Antifungal activity of allylamines on Epidermophyton floccosum: scanning electron microscopy study

The action of allylamine antifungal agents on Epidermophyton floccosum was studied using scanning electron microscopy. After 7 days of culture on Sabouraud dextrose agar, Epidermophyton floccosum samples were brought in contact with concentrations of 0.2 and 2 g ml^–1 and 0.01 and 0.1 g ml^–1 of naftifine and terbinafine, respectively. Lesions observed after 24 h, 3 and 7 days of contact were mainly on the structure and rigidity of the mycelial and macroconidial wall. They were characterized by hyphal ballooning and twisting and by apical bulbous bulges. Deterioration of macroconidia was characterized by wall exfoliation. The intensity of the deterioration depended on the dose and only slightly on the length of time that the sample and the antifungal drug were in contact.     

The growth of Epidermophyton floccosum and E. stockdaleae at different temperatures

The ability of 17 strains of genus Epidermophyton (15 strains belonging to Epidermophyton floccosum, one to E. floccosum var. nigricans and one to E. stockdaleae) to grow at different temperatures (4 °C, 25 °C, 28 °C, 31 °C, 34 °C, 37 °C and 40 °C) was stated.The strains were inoculated on Sabouraud Dextrose Agar and regularly controled over a period of 14 days when the plates were incubated at 25 °C, 28 °C, 31 °C, 34 °C, 37 °C and 40 °C, and over a period of 70 days when the temperature was 4 °C. The optimal growth of E. floccosum was observed at 28 °C and 31 °C, and no signs of growth were recorded neither at 4 °C nor at 40 °C. The optimal development of E. stockdaleae was observed at 25 °C and 28 °C. This species grew from 4 °C to 31 °C.     

Nocardia brasiliensis: in vitro and in vivo growth response to steroid sex hormones

As actinomycetoma is more frequent in males than in females, the possibility that hormones might modify theNocardia brasiliensis growth and the course of experimental actinomycetoma was explored. FiveN. brasiliensis strains were grown on Sabouraud agar containing estradiol, progesterone or testosterone, in 3 different concentrations. Colony diameters were measured weekly for 7 weeks.N. brasiliensis strains were also grown in Sabouraud broth containing hormones. Glucose concentration was measured weekly for 6 weeks. Finally, experimental actinomycetoma was produced in male and female hormone-treated mice. Invasion rate, plantar pad diameter and positive retrocultures were assessed. In vitro experiments showed that progesterone and testosterone inhibitN. brasiliensis growth, manifested by lower colony diameters and greater glucose concentrations. In vivo experiments demonstrated that estradiol limits actinomycetoma development. Progesterone and testosterone induced greater diameters of inoculated plantar pads and greater invasion rates with greater positive culture numbers than estradiol. Results partially explain the resistance of females to actinomycetoma.     

Comparison of coal solubilization by bacteria and fungi

Coal-solubilizing agents produced byTrametes versicolor, Phanerochaete chrysosporium, Aspergillus sp., a bacterial consortium, and a bacterial isolate,Arthrobacter sp., from that consortium were compared in terms of pH dependence, thermostability, molecular mass, mechanism of action, and product diversity. The thermostability and low molecular weights exhibited by the coal-solubilizing agents indicated a non-enzymatic mechanism of action. Competition studies using cupric copper indicated that coal solubilization by these agents involved metal chelation. Results demonstrated that oxalate could account for some but not all of the coal solubilization observed forT.versicolor andP.chrysosporium. The very low levels of oxalate detected inAspergillus sp. and the bacterial cultures indicated that oxalate is not an important factor in coal solubilization by these microbes. When subjected to gel permeation chromatography, the soluble coal products generated by each microbial coal-solubilizing agent yielded unique molecular mass profiles suggesting substantial product diversity. Such diversity increases the possibility of identifying potentially valuable compounds and extending the commercial utilization of coal.Abbreviations A₄₅₀, A₂₆₀ absorbances respectively at 450 nm and 260 nm - CSA coal-solubilizing agent - CSU coal-solubilizing unit - GPC gel permeation chromatography - MEA malt extract agar - PDA potato dextrose agar - SDA Sabouraud dextrose agar - SDB Sabouraud dextrose broth - SEM standard error of the mean - Tris tris(hydroxymethyl)aminomethane - TSA trypticase soy agar - TSB trypticase soy broth     

Simple rapid identification of Candida albicans with emphasis on the differentiation between Candida albicans and Candida stellatoidea

Summary Subcultures ofC. albicans, made from Sabouraud agar, grown at room temperature for 48 hours, were inoculated into a 10 times saline dilution of Sabouraud liquid medium and left in the incubator for 45–60 minutes at 37° C, transferred to corn meal agar plates and incubated at 37° C for 18–24 hours.Small portions of the surface agar containing the yeasts from these plates were pressed under cover glasses and examined under the oil immersion lens.Under these conditions,C. albicans cultures were observed to produce only yeast-like cells, whereasC. stellatoidea cultures contained predominantly abundant, long, thin mycelia.     

Variability of biochemical characteristics in strains of Trichophyton mentagrophytes

The characteristics of 23 strains of Trichophyton mentagrophytes isolated from clinical samples, were studied. Sabouraud agar was the most adequate medium to inhance at their utmost the different taxonomical features. Variability of enzymatic reactions in strains of T. mentagrophytes was observed.     

Isolation of fluconazole-tolerant micromycetes onto different cultivation media

On studying the dependence of quality and/or quantity of environmental fluconazole-tolerant micromycetes on culture mediumin vitro, the highest number of isolates was obtained in Sabouraud agar (SA) without fluconazole (59 CFU/mg of soil sample at 25°C and 31 CFU/mg at 37°C). In the case of SA, fluconazole inhibited the growth of micromycetes to 83%. Significant differences were found between the average concentration of CFU/mg for six cultivation media without and with fluconazole (70 mg/L) at 25 and 37°C. The genusAspergillus (A. flavus, A. fumigatus, aspergilli fromA. glaucus andA. niger group,A. nidulans, A. versicolor, A. ustus), Scopulariopsis brevicaulis andPenicillium sp. were the most common isolates. Sabouraud agar combined with malt extract agar, dichloran-18% glycerol agar and potato-dextrose agar were the most suitable for isolation of fluconazole-tolerant fungi.     

Outbreaks of phaeohyphomycosis in the chinook salmon (Oncorhynchus tshawytscha) caused by Phoma herbarum

Phoma herbarum has been associated with two outbreaks of systemic mycosis in hatchery-reared chinook salmon (Oncorhynchus tshawytscha) fingerlings. Affected fish exhibited abnormal swimming behavior, exophthalmia, multiple rounded areas of muscle softening, protruded hemorrhagic vents, and abdominal swelling. In all affected fish, swimbladders were filled with whitish creamy viscous fungal mass, surrounded by dark red areas in swimbladder walls, kidneys, and musculature. Clinical and histopathological examinations suggest that the infection may have started primarily in the swimbladder and then spread to the kidneys, gastrointestinal tract, and surrounding musculature. Consistent microscopical findings included broad septate branched fungal hyaline hyphae, 5–12 μm in diameter within the swimbladder, stomach, and often within and adjacent to blood vessels. Profuse growths of woolly brown fungal colonies were obtained from swimbladders and kidneys on Sabouraud medium. On corn meal agar the formation of pycnidia, characteristic of Phoma spp., was detected within 10 days of incubation. Morphological and molecular analyses identified this fungus as Phoma herbarum. This report underscores systemic fungal infections as a threat to raceway-raised salmon.     

Influence of fungicides and insecticides on the entomogenous fungus Metarhizium anisopliae a pathogen of the vine weevil,Otiorhynchus sulcatus

In the laboratory, the fungicides chlorothalonil and zineb prevented germination of Metarhizium anisopliae conidia when incorporated into Sabouraud dextrose agar (SDA) at the commercial concentration (based on the manufacturers’ recommended rates for horticultural crops). Twelve other fungicides and six insecticides had no effect on spore germination when applied at the same rate. Mycelial growth of M. anisopliae on SDA plates containing the recommended rate of all the pesticides (except propamocarb) was reduced compared with SDA alone. Two fungicides, benomyl and carbendazim, totally inhibited growth at 0.1 times the recommended rate. Growth was also completely prevented by the fungicides etridiazole, triforine and zineb, and the insecticides dichlorvos and hostathion, at 10 times the recommended rate. In a glasshouse experiment, a prophylactic drench of M. anisopliae conidia reduced vine weevil (Otiorhynchus sulcatus) populations on Impatiens plants by 88%. This level of control was not significantly reduced by subsequent application (7 days after egg infestation) of any of the pesticides at the recommended concentration. Larval control in pots treated with M. anisopliae plus any one of the 12 fungicides and four insecticides examined, ranged from 82% to 98%. The insecticide diazinon applied alone reduced larval numbers by 100%. Two other insecticides, dichlorvos and cypermethrin, and the fungicide pyrazaphos, also reduced weevil populations by over 50%. These experiments demonstrate the limitations of laboratory based in vitro screening programmes for assessing the chemical compatibility of M. anisopliae.     

<Emphasis Type="Italic">Trichophyton Rubrum</Emphasis> with the ‘<Emphasis Type="Italic">raubitschekii</Emphasis>’ Morphotype: The First Report from Turkey

In modern taxonomy, Trichophyton rubrum with the ‘raubitschekii’ morphotype differs from the common strains of T. rubrum morphologically, physiologically, and clinically, despite genetic homogeneity. In this paper, we report the first autochthonous Turkish cases in a 24-year-old woman and her 26-year-old fiancé with tinea cruris and tinea manuum, respectively. The isolates were proven to be T. rubrum with the ‘raubitschekii’ morphotype by their morphological and physiological characteristics including the following: (1) granular colony surface, (2) profuse macroconidia and microconidia, (3) strongly positive urease activity after 3 days, (4) non-perforating hair, (5) restricted growth and producing no alkaline reaction on bromocresol purple-milk solids-glucose agar, and (6) restricted growth and producing no conidia on 3–9% NaCl-supplemented Sabouraud agar. The internal transcribed spacer (ITS) regions of both isolates were sequenced, and the assembled DNA sequences were examined using the Basic BLAST (nucleotide–nucleotide) software of the National Center for Biotechnology Information web database. Actually, the lack of epidemiological data on this variety is notable, and the actual prevalence is unknown. Otherwise, routine methods misidentified the strains as T. rubrum or unrecognized cases were underreported.