Sidney A. Crow

Adherence of Candida albicans to Silicone Induces Immediate Enhanced Tolerance to Fluconazole

ABSTRACT Wild-type and efflux pump-deficient cells of Candida albicans adhering to silicone were compared with planktonic cells by flow cytometry for their relative resistance to fluconazole (FCZ). Flow cytometry data on cells carrying a fusion of green fluorescent protein to efflux pump promoters confirmed that enhanced tolerance of attached cells to FCZ was due in part to increased expression of CaMDR1 and CDR1 promoters. Within 2 h of their attachment to silicone, the adherent cells demonstrated levels of FCZ tolerance shown by cells from 24-h biofilms. Following their mechanical detachment, this subset of cells retained a four- to eightfold increase in tolerance compared with the tolerance of planktonic cells for at least two generations. Enhanced efflux pump tolerance to FCZ appeared to be induced within the initial 15 min of attachment in a subset of cells that were firmly attached to the substrata.

Metabolism of aromatic hydrocarbons by yeasts

Six yeasts were examined for their ability to metabolize naphthalene, biphenyl and benzo(a)pyrene. All of the organisms tested oxidized these aromatic hydrocarbons. Candida lipolytica oxidized naphthalene to 1-naphthol, 2-naphthol, 4-hydroxy-1-tetralone and trans-1,2-dihydroxy-1,2-dihydronaphthalene. The major metabolite was 1-naphthol. C. lipolytica oxidized biphenyl to produce 2-, 3-, and 4-hydroxybiphenyl, 4,4′-dihydroxybiphenyl and 3-methoxy-4-hydroxybiphenyl. 4-Hydroxybiphenyl was the predominant metabolite formed. C. lipolytica oxidized benzo(a)pyrene to 3-hydroxybenzo(a)pyrene and 9-hydroxybenzo(a)pyrene. Metabolites were isolated and identified by absorption spectrophotometry, mass spectrometry and thin-layer, gasliquid and high-pressure liquid chromatography. Where possible the structures of these metabolites were confirmed by comparison with authenic compounds.

Initial oxidative and subsequent conjugative metabolites produced during the metabolism of phenanthrene by fungi

Three filamentous fungi were examined for the ability to biotransform phenanthrene to oxidative (phase I) and conjugative (phase II) metabolites. Phenanthrene metabolites were purified by high-performance liquid chromatography (HPLC) and identified by UV/visible absorption, mass, and1H NMR spectra.Aspergillus niger ATCC 6275,Syncephalastrum racemosum UT-70, andCunninghamella elegans ATCC 9245 initially transformed [9-14C]phenanthrene to produce metabolites at the 9,10-, 1,2-, and 3,4- positions. Subsequently, sulfate conjugates of phase I metabolites were formed byA. niger, S. racemosum, andC. elegans. Minor glucuronide conjugates of 9-phenanthrol and phenanthrenetrans-9,10-dihydrodiol were formed byS. racemosum andA. niger, respectively. In addition,C. elegans produced the glucose conjugates 1-phenanthryl β-d-glucopyranoside and 2-hydroxy-1-phenanthryl β-d-glucopyranoside, a novel metabolite. [9-14C]Phenanthrene metabolites were not detected in organic extracts from biotransformation experiments with the yeasts,Candida lipolytica 37-1,Candida tropicalis ATCC 32113, andCandida maltosa R-42.

Fungal colonization of air filters from hospitals.

Air filters of various types, selected on the basis of discoloration, were collected from the primary and secondary filter banks of the heating, ventilating, and air-conditioning systems in seven hospitals in the eastern United States and examined with direct microscopy for fungal colonization. Microscopic observations and culture results showed that filters from five of the hospitals were colonized with fungi including species of Acremonium, Alternaria, Aspergillus, Cladosporium, Epicoccum, Penicillium, and Rhinocladiella, and a Beauveria-like fungus. Several of these commonly airborne species, e.g., Epicoccum purpurescens (syn. E. nigrum) and Rhinocladiella sp., had not been previously reported to colonize (with conidiogenesis) air filters.

Fungal colonization of air filters for use in heating, ventilating, and air conditioning (HVAC) systems

SummaryNew and used cellulosic air filters for HVAC systems including those treated with antimicrobials were suspended in vessels with a range of relative humidities (55–99%) and containing non-sterile potting soil which stimulates fungal growth. Most filters yielded fungi prior to suspension in the chambers but only two of 14 nontreated filters demonstrated fungal colonization following use in HVAC systems. Filters treated with antimicrobials, particularly a phosphated amine complex, demonstrated markedly less fungal colonization than nontreated filters. In comparison with nontreated cellulosic filters, fungal colonization of antimicrobial-treated cellulosic filters was selective and delayed.

Fungal Colonization of Air Filters and Insulation in a Multi-Story Office Building: Production of Volatile Organics

Abstract. Secondary air filters in the air-handling units on four floors of a multi-story office building with a history of fungal colonization of insulation within the air distribution system were examined for the presence of growing fungi and production of volatile organic compounds. Fungal mycelium and conidia of Cladosporium and Penicillium spp. were observed on insulation from all floors and both sides of the air filters from one floor. Lower concentrations of volatile organics were released from air filter medium colonized with fungi as compared with noncolonized filter medium. However, the volatiles from the colonized filter medium included fungal metabolites such as acetone and a carbonyl sulfide-like compound that were not released from noncolonized filter medium. The growth of fungi in air distribution systems may affect the content of volatile organics in indoor air.

Growth and survival of Fusarium solani-F. oxysporum complex on stressed multipurpose contact lens care solution films on plastic surfaces in situ and in vitro.

Purpose: To analyze factors implicating the association of ReNu with MoistureLoc (ReNu ML) multipurpose contact lens solution (MPS) with the increased incidence of Fusarium keratitis. Methods: Used contact lens cases with and without contact lenses and MPS containers were collected from patients with confirmed or possible Fusarium keratitis. Direct microscopy including transparent adhesive tape preparations and swab cultures were used to determine fungal colonization. Survival and growth of selected isolates of Fusarium spp. in drying MPS on plastic surfaces were determined by microscopy and recoverable colony counts on enriched agar. Results: Discrete regions of fungal colonization, including occasional microcycle conidiation and chlamydospore formation, were observed on the surfaces of contact lens cases and, less often, on solution containers that had been used by patients with Fusarium keratitis associated with the use of ReNu ML. Isolates provisionally grouped with the F. solani-F. oxysporum complex were inhibited by fresh MPS in original solution containers and contact lens cases, but survived in stressed (drying) films of MPS, particularly ReNu ML. These in vitro test results were similar to the direct in situ observations of the materials from patients. Conclusions: Selective, rapid growth and survival of cells of the F. solani-F. oxysporum complex on plastic surfaces, particularly of contact lens cases with stressed ReNu ML films, may explain, in part, the recent Fusarium keratitis outbreak.

The Occurrence and Persistence of Mixed Biofilms in Automobile Air Conditioning Systems

Abstract. Twelve automobile air conditioner systems from six manufacturers and three countries, selected mostly because of complaints of unpleasant odors in the passenger compartment, were examined for microbial growth by direct microscopy and enrichment culture. Mixed populations of fungi and bacteria (with occasional protozoa) were observed in biofilms in at least some of the components from all used units. The aluminum heat exchanger fins from ten evaporators demonstrated bacterial biofilms that yielded Methylobacterium mesophilicum. Penicillium viridicatum colonized components from four units. These bacteria and fungi were recoverable repeatedly from these units during ‘dry’ storage of up to 27 months. This report associates a bacterial-fungal community with disagreeable air quality in some automobiles.

Fusarium keratitis and contact lens wear: facts and speculations

Over the past several decades mycotic keratitis has been considered a rare sequel to hydrogel contact lens wear. In 2005--2006 an upswing in the incidence of Fusarium keratitis was associated with a disproportionate use of one multipurpose contact lens solution (MPS, ReNu with MoistureLoc, Bausch & Lomb, Rochester, NY). The MPS, as manufactured and marketed, was sterile and met regulatory guidelines for antimicrobial activity. A multivariant interaction of poor hygienic practices and the contact lens paraphernalia were associated with a mostly selective contamination in or on the lens storage case by members of the F. solani/F. oxysporum species complexes from the environment of the user. A decline of the anti-fusaria properties of the MPS in the lens case appeared related to its dissociation from drying, or dilution and the potential for sorption of antimicrobial solution components (e.g., alexidine) to various hydrogel lenses. These factors and capacities of the fusaria for rapid amplification by microcycle conidiation, production of dormant resistant cells, and potential for attachment and penetration of hydrogel lenses, were linked to the occasional selective fungal survival and growth during storage of the lens in MPS. Lack of a manual rubbing-cleaning step in the MPS disinfection process was considered a risk factor for keratitis.

Volatile organic compounds associated with microbial growth in automobile air conditioning systems.

Volatile organic compounds from Penicillium viridicatum and Methylobacterium mesophilicum growing on laboratory media and on component materials of automobile air conditioners were analyzed with gas chromatography and mass spectrometry. P. viridicatum produced compounds such as 4-methyl thiazole, terpenes and alcohols, whereas M. mesophilicum produced dimethyl disulfide, dimethyl trisulfide, and chlorophenol with growth on laboratory media. In comparison with laboratory media, fewer volatiles were detected from colonized foam insulation materials. Biofilms of M. mesophilicum on aluminum evaporator components produced mainly dimethyl disulfide. These biofilms, after inoculation with P. viridicatum, produced offensive smelling alcohols and esters such as 2-methyl propanol, 3-penten-2-ol, and the ethyl ester of butanoic acid. The moisture and substrates innate to the automobile air conditioning systems provided an environment suitable for microbial biofilm development and odor production. Reduction of retained moisture in the air conditioning system coupled with use of less susceptible or antimicrobial substrates are advised for remediation of the noxious odors.