Blastomycosis diagnosed in a nonhyperendemic area.

INTRODUCTION: Blastomycosis, caused by the dimorphic fungus Blastomyces dermatitidis, is hyperendemic in northern Wisconsin and is unevenly distributed in the rest of the state and contiguous Minnesota. Clinical presentation of this illness has been characterized by localized outbreaks and sporadic cases in endemic areas. METHODS: Using ICD-9 CPT codes, we queried our electronic health record system to identify cases of blastomycosis diagnosed at Gundersen Health System over a 32-year period. Gundersen serves a region outside the hyperendemic area of Wisconsin. Records so identified were reviewed for demographic and clinical features. We attempted to interview patients with a blastomycosis diagnosis from 2002 through 2006. Blastomycosis data also were collected from the states of Wisconsin and Minnesota from 2002 through 2006 and assessed for trends. RESULTS: Thirty-six patients had blastomycosis diagnoses at Gundersen Health System during the study period, as identified by ICD-9 code. Of these, 10 were excluded from further review because they were either miscoded or the code indicated a previous diagnosis. The remaining 26 patients were included in the study. Premorbid conditions included diabetes (38%) and smoking (62%). The mean time from onset of symptoms to the first laboratory specimen positive for B dermatitidis was 51 days. Notably, 73% of these patients were treated initially for bacterial pneumonia. The incidence of blastomycosis in Wisconsin in the review period was 2.0 per 100,000, and the rate in Minnesota was 0.5 per 100,000. Based on the census data in Gundersen Health System's 19-county service area, the incidence of blastomycosis is 0.17 cases per 100,000. CONCLUSION: In this review of blastomycosis cases diagnosed outside the hyperendemic area of northern Wisconsin, diagnosis was often delayed, and 4 patients whose infections might have been treatable died. Although uncommon, blastomycosis needs to be considered in the differential diagnosis in areas outside the hyperendemic area.

Molecular phylogeny and morphology reveal three new species of Cantharellus within 20 m of one another in western Wisconsin, USA.

Three new species, Cantharellus phasmatis, Cantharellus flavus and Cantharellus spectaculus, all previously considered Cantharellus cibarius, are described in this study. The circumscription of these three species from C. cibarius and other Cantharellus species is supported by morphological differences and nuclear DNA sequence data (nLSU, ITS, TEF1). All were found under Quercus spp. in a small plot in Hixon Forest Park in La Crosse, Wisconsin, emphasizing the need for further taxonomic study of even common and conspicuous genera in North America. In addition, a review of the current state of C. cibarius sensu lato systematics is presented, including a review of the recent elevation of C. cibarius var. roseocanus to the species rank. Taxonomic descriptions and photographs are provided for the newly described species.

Antibacterial spirobisnaphthalenes from the North American cup fungus Urnula craterium.

Urnucratins A-C (1-3), which possess an unusual bisnaphthospiroether skeleton with one oxygen bridge and one C-C bridge and represent a new subclass of bisnaphthalenes, were isolated from the North American cup fungus Urnula craterium. Their structures, including absolute configurations, were determined by means of HRMS, NMR, and quantum chemical CD calculations. Urnucratin A (1) was found to be active against methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecium, and Streptococcus pyogenes with MIC values of 2, 1, and 0.5 µg/mL, respectively.

Taxonomic revision of true morels (Morchella) in Canada and the United States.

Recent molecular phylogenetic studies have revealed the existence of at least 50 species of Morchella worldwide and demonstrated a high degree of continental endemism within the genus. Here we describe 19 phylogenetic species of Morchella from North America, 14 of which are new (M. diminutiva, M. virginiana, M. esculentoides, M. prava, M. cryptica, M. frustrata, M. populiphila, M. sextelata, M. septimelata, M. capitata, M. importuna, M. snyderi, M. brunnea and M. septentrionalis). Existing species names (M. rufobrunnea, M. tomentosa, M. punctipes and M. angusticeps) are applied to four phylogenetic species, and formal description of one species (M. sp. "Mel-8") is deferred pending study of additional material. Methods for assessing morphological features in Morchella are delineated, and a key to the known phylogenetic species of Morchella in North America is provided. Type studies of M. crassistipa, M. hotsonii, M. angusticeps and M. punctipes are provided. Morchella crassistipa is designated nomen dubium.

Screening a mushroom extract library for activity against Acinetobacter baumannii and Burkholderia cepacia and the identification of a compound with anti-Burkholderia activity.

BACKGROUND: Acinetobacter baumannii and species within the Burkholderia cepacia complex (BCC) are significant opportunistic bacterial pathogens of humans. These species exhibit a high degree of antibiotic resistance, and some clinical isolates are resistant to all currently available antimicrobial drugs used for treatment. Thus, new drugs are needed to treat infections by these species. Mushrooms could be a potential source for new drugs to treat A. baumannii and BCC infections. METHODS: The aim of this study was to screen a library of crude extracts from 330 wild mushrooms by disk diffusion assays for antibacterial activity against A. baumannii and Burkholderia cepacia in the hope of identifying a novel natural drug that could be used to treat infections caused by these species. Once positive hits were identified, the extracts were subjected to bioassay-guided separations to isolate and identify the active drug molecules. MICs were performed to gauge the in vitro activity of the purified compounds. RESULTS: Only three crude extracts (0.9%) had activity against A. baumannii and B. cepacia. Compounds from two of these extracts had MICs greater than 128 microg/ml, and further analyses were not performed. From the third extract, prepared from Leucopaxillus albissimus, 2-aminoquinoline (2-AQ) was isolated. This compound exhibited a modest MIC in vitro against strains from nine different BCC species, including multi-drug resistant clinical isolates (MIC = 8-64 microg/ml), and a weak MIC (128 microg/ml) against A baumannii. The IC50 against a murine monocyte line was 1.5 mg/ml. CONCLUSION: The small number of positive hits in this study suggests that finding a new drug from mushrooms to treat Gram-negative bacterial infections may be difficult. Although 2-AQ was identified in one mushroom, and it was shown to inhibit the growth of multi-drug resistant BCC isolates, the relatively high MICs (8-128 microg/ml) for both A. baumannii and BCC strains suggests that 2-AQ is not suitable for further drug development in its current form.

Antibacterial compounds from mushrooms II: lanostane triterpenoids and an ergostane steroid with activity against Bacillus cereus isolated from Fomitopsis pinicola.

Anti- Bacillus cereus bioassay-guided fractionation of a crude extract of the American mushroom, Fomitopsis pinicola, was performed using thin-layer chromatography, Sephadex LH-20 column chromatography, and preparative-scale HPLC. Five lanostane triterpenoids (1-5) and one ergostane steroid (6) were isolated and identified. Compound 1 is a new lanostane triterpenoid, and its structure was determined using 1D and 2D NMR experiments, HR-MS, and physical data. Each of the purified compounds (1-6) was tested for antibacterial activity against B. cereus using standard MIC assays. Compounds 1-6 had MIC values of 32, 16, 32, 32, 128, and 64 microg/mL, respectively.

Antibacterial compounds from mushrooms I: a lanostane-type triterpene and prenylphenol derivatives from Jahnoporus hirtus and Albatrellus flettii and their activities against Bacillus cereus and Enterococcus faecalis.

Antibacterial bioassay-guided fractionation of two American mushroom species, Jahnoporus hirtus and Albatrellus flettii, led to the isolation and identification of their major antibacterial constituents: 3,11-dioxolanosta-8,24( Z)-diene-26-oic acid (1) from J. hirtus and confluentin (2), grifolin (3), and neogrifolin (4) from A. flettii. Compound 1 is a new lanostane-type triterpene. All purified compounds were evaluated for their ability to inhibit the growth of Bacillus cereus and Enterococcus faecalis using standard MIC assays. Compounds 1- 4 demonstrated MIC values of 40, 20, 10, and 20 microg/mL, respectively, against B. cereus and MIC values of 32, 1.0, 0.5, and 0.5 microg/mL, respectively, against E. faecalis. Thus, one novel compound and three others were shown to possess antimicrobial activities against these gram-positive bacteria employed as surrogates for more virulent and dangerous pathogens.

Identification of Lacrymaria velutina (Pers. Ex Fr.) Konrad & Maubl. from Micheon-myeon, Jinju-city, Korea.

We identified Lacrymaria velutina of the Coprinaceae in Korea. The unusually large and sturdy fruiting body, fibrillose to fibrillose-scaly cap and stalk without a volva with an obscure superior hairy ring zone or hairy annulus, and blackish brown, warted spores distinguished this species from closely related Psathyrella species. An illustrated account of the microscopic traits is presented. Fruiting bodies with obtusely hemispherical caps, 2.5~6 cm, becoming convex with age; surface dry, densely fibrillose-scaly with split margin; stipe, 4.5~6 cm, equal, hollow, fibrillose, dry, whitish above the superior ring zone, light brown below; crowded gills, adnexed, dark black at maturity. Pileipellis typically cellular with the gill edge appearing white and beaded. Blackish brown basidiospores that discolor in concentrated sulfuric acid. Spores elliptical, warted, 9~11 × 6~8 µm, with prominent snout-like germpores. Cheilocystidia abundant, 57~68 × 19~25 µm, and narrowly elongated clavate, often clustered in threes or fours. Pleurocystidia rarely present, 45~47.5 × 12~13 µm, and clavate to utriform. This trait distinguishes our sample as L. velutina from other Psathyrella spp. of the Coprinaceae, which have smooth spores. This taxon was clarified by the observation that Psathyrella spores fade in concentrated sulfuric acid. A molecular phylogenetic study revealed that our specimen was Lacrymria velutipes, which is closely related to Lacrymaria lacrymabunda. Moreover, those two species are clearly distinguishable from other Psathyrella species, which agreed with the morphologically distinctive traits described above. We believe that this is the first report of this taxon, which has not been described in Korea.

Ectomycorrhizal characterization of an American chestnut (Castanea dentata)-dominated community in Western Wisconsin.

Circa 1900, a farmer from the eastern US planted 11 American chestnut (Castanea dentata) seeds on a newly established farm near West Salem in western Wisconsin. These trees were very successful, producing a large stand of over 6,000 trees. Since this area is well outside the natural range of chestnut, these trees remained free from chestnut blight until 1987. In the West Salem stand, chestnuts are the dominant species of a mixed forest community, reminiscent of the chestnut-oak ecosystems of pre-1900 Appalachia. To identify putative mycorrhizal associates of chestnut in this unique forest, our approach was twofold: (1) an extensive fruiting body survey was conducted for four seasons that yielded approximately 100 putative mycorrhizal species and (2) a belowground molecular approach was used to generate DNA sequences of the internal transcribed spacer region from ectomycorrhizae. Unexpectedly, chestnut did not appear to be the dominant underground ectomycorrhizal-forming plant species. This study highlights the need to identify the plant host species when conducting belowground molecular-based surveys and provides preliminary identification of ectomycorrhizal fungi associated with a disjunct stand of American chestnut.

White-rot fungi demonstrate first biodegradation of phenolic resin.

Phenolic resins, phenol-formaldehyde polymers previously thought to be nonbiodegradable, are produced at an annual rate of 2.2 million metric tons in the United States for many industrial and commercial applications. Three independent lines of evidence established their biodegradability with the white-rot fungus Phanerochaete chrysosporium. Chromatic transformation of growth medium (yellow to pink) indicated initial biodegradation of the resin 3 days after inoculation. A degradation product, 13C-labeled phenol, was detected with gas chromatography-mass spectroscopy. Scanning electron micrographs revealed physical evidence of degradation. This is the first demonstrated biodegradation of these phenol-formaldehyde polymers and stands as a platform for investigation into bioremediation and biorecycling of phenolic resins.