Yarrowia lipolytica is an ascomycetous yeast commonly found in meats, including sausages, and dairy products, especially cheeses. Its anamorph is Candida lipolytica . This yeast is a valued component of starter cultures for some sausages and cheeses, but it causes spoilage in other products. It is used for many biotechnological applications, including the production of citric acid, enzymes, and pharmaceuticals, and for the bioremediation of wastes.
Despite the use of refrigeration, improved packaging, adsorbents, and ethylene receptor blockers, on average, nearly 40% of all fruits and vegetables harvested in the US are not consumed. Many plant products, especially fruit, continue to ripen after harvesting, and as they do so, become increasingly susceptible to mechanical injury, resulting in increased rot. Other plant products during transportation and storage are susceptible to chill injury (CI). There is a real need for products that can delay ripening or mitigate the effects of CI, yet still permit full ripeness and quality to be achieved. Preliminary results are discussed where catalyst derived from cells of Rhodococcus rhodochrous DAP 96253, grown under conditions that induced high levels of nitrile hydratase, were able to extend the ripening and thus the shelf-life of selected climacteric fruits (banana, avocado, and peach). A catalyst, when placed in proximity to, but not touching, the test fruit delayed the ripening but did not alter the final ripeness of the fruit tested. Organo-leptic evaluations conducted with control peaches and with peaches exposed to, but not in contact with, the catalyst showed that the catalyst-treated peaches achieved full, natural levels of ripeness with respect to aroma, flavor, sweetness, and juice content. Furthermore, the results of delayed ripening were achieved at ambient temperatures (without the need for refrigeration).
Microcycle conidiation and microniche colonization by aspergilli was observed in-situ on various indoor construction and finishing materials. Microcycle conidiation, direct conidiogenesis from a conidium or spore with minimal intervening hyphal development, for several decades has been considered a survival mechanism during stress for a variety of moulds. Adhesive transparent tape mounts and bulk materials from various indoor materials, including air filters from hospitals and healthcare institutions, were transported to the laboratory for light microscopic and scanning electron microscopic observations. Additional materials were held in moist chambers over nonsterile soils and examined periodically for fungal development. Microcycle conidiation was observed usually in areas of sparse fungal development, mostly in association with isolations of members of the Aspergillus flavus-, A. versicolor-, A. niger groups. Branched conidiophores and medusa heads, more often associated with colonization by Eurotium spp., were observed on some preserved woods. These conidiogenesis processes might be factors in the survival and blooms of indoor aspergilli.
Sixteen commercial products for use in automobile air-conditioning systems (ACS), most designated for abatement of malodors presumably of microbial origin, were examined for their potential to inhibit attachment and to detach cells of the Gram-negative bacterium Serratia marcescens on aluminum sections. Numbers of attached cells were appreciably reduced (>60%) following immersion in three alcohol-type and two acrylic-coating-type products. Several products had essentially no effect on the attached cells. Most of the products indicated for alleviation of associated microbial odors from ACS provided only short-term effects. When products were coated onto aluminum prior to exposure to the cells, water-insoluble coatings appeared to provide more consistent inhibition of primary adherence of S. marcescens. The differences in degrees of primary adherence of a selected strain of S. marcescens to variously treated aluminum provided a rapid and reproducible assessment of potential antimicrobial efficacy of ACS products.
The rapid rise of energy costs in the 1970's and early 1980's brought about a number of energy conservation measures which are still in use today. Changes made in heating, ventilating and air conditioning (HVAC) systems increased the recirculation of conditioned air within buildings, while leakage of air through the building shell was sharply reduced. Air movement in modern buildings is typically under the control of centrally operated HVAC systems which do not allow for local control of airflow in particular areas by the occupants. Problems with the quality of indoor air in the work environment have resulted from some of these measures and collectively many of these phenomena have been referred to as the Sick Building Syndrome (SBS). SBS is a complex of symptoms, including eye and mucous membrane irritation, headache, cough, dizziness, chest tightening, fatigue and general malaise, which occur more frequently than would normally be expected (greater than 20%) among the occupants of a building.
Automobile air conditioning systems (ACS) might be considered an extreme environment for many microorganisms. Organisms surviving and proliferating in these systems may be presented with temperature changes ranging from subzero to over 140°F, water activity from saturation to dryness and a nutrient complexity including varying levels of hydrocarbons. Microbial communities develop in ACS and sometimes proliferate to the extent of massive colonization and production of objectionable odors.In a few instances microorganisms emanating from ACS have been associated with hypersensitivity pneumonitis and other allergic reactions. Previously we demonstrated that foam insulation and glues, in particular, on ACS insulations were colonized by fungi such as Aspergillus, Aureobasidium, Cladosporium, and Penicillium. Such fungi often are implicated in colonization of similar substrates in buildings categorized with the sick building syndrome.A total of 42 ACS from the United States, Europe and Central America have been examined by cabin air sampling culture swab, direct microscopy or a combination of these techniques.
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.
Air samples and swab samples of the air conditioning vents were collected from 29 automobiles in the metropolitan region of Atlanta, GA, and cultured for fungi. Among the fungi observed, species of Acremonium, Aspergillus, Alternaria, Aureobasidium, Cladosporium, and Penicillium were in the highest densities. Transparent adhesive tape imprints, SEM observations, and enrichment culture of components of five systems demonstrated fungal hyphae on the metal surfaces and within the matrix of various insulation materials. The evaporator removed from one automobile because of a series of complaints of noxious odors was densely colonized by Penicillium viridicatum. The amplification of known allergenic and odor-producing fungi occurred within the automobile air conditioning systems.
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.
Complaints characteristic of those for sick building syndrome prompted mycological investigations of a modern multi-story office building on the Gulf coast in the Southeastern United States (Houston-Galveston area). The air handling units and fiberglass duct liner of the heating, ventilating and air conditioning system of the building, without a history of catastrophic or chronic water damage, demonstrated extensive colonization withPenicillium spp andCladosporium herbarum. Although dense fungal growth was observed on surfaces within the heating-cooling system, most air samples yielded fewer than 200 CFU m−3. Several volatile compounds found in the building air were released also from colonized fiberglass. Removal of colonized insulation from the floor receiving the majority of complaints of mouldy air and continuous operation of the units supplying this floor resulted in a reduction in the number of complaints.
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.
New 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.
Materials being used or considered for use in space flight were examined for their susceptibility to fungal colonization. The materials included soft goods (clothing) and insulation and fabrication products such as Velcro® attachments and elastic cord binders. Materials were exposed for at least 28 days in a highhumidity chamber colonized with over 50 species of fungi, including those species recommended for determining recalcitrance of materials to fungal biodegradation. At least nine of 25 products demonstrated extensive microscopic colonization by fungi, mostly byAcremonium obclavatum. Challenge procedures that rely on observations with the unaided eye, or 40×magnification of growth by a restricted number of fungal species with a cellulosic substrate as a positive control, are insufficient for determining the resistance of synthetic substrates to fungal colonization.
The number of fungal species colonizing thermal and acoustic fiberglass insulations used in heating, ventilation, and air conditioning (HVAC) systems was fewer than that obtained from initial direct culture of these insulations. The colonization, determined by the microscopic observation of conidiophores with conidia, was primarily of acrylic-latex-facing material, but eventually the fungi permeated the fiberglass matrix. Isolates ofAspergillus versicolor were most often obtained from non-challenged insulation, whereasAcremonium obclavatum appeared to be the primary colonizing fungus in high-humidity (>90%) challenge chambers. At a lower humidity (about 70%)Aspergillus flavus was one of the more prominent fungi. Not all duct liner samples were equally susceptible to colonization and duct board appeared relatively resistant to colonization.
The trans-dihydrodiols produced during the metabolism of phenanthrene by Cunninghamella elegans, Syncephalastrum racemosum, and Phanerochaete chrysosporium were purified by high-performance liquid chromatography (HPLC). The enantiomeric compositions and optical purities of the trans-dihydrodiols were determined to compare interspecific differences in the regio- and stereoselectivity of the fungal enzymes. Circular dichroism spectra of the trans-dihydrodiols were obtained, and the enantiomeric composition of each preparation was analyzed by HPLC with a chiral stationary-phase column. The phenanthrene trans-1,2-dihydrodiol produced by C. elegans was a mixture of the 1R,2R and 1S,2S enantiomers in variable proportions. The phenanthrene trans-3,4-dihydrodiol produced by P. chrysosporium was the optically pure 3R,4R enantiomer, but that produced by S. racemosum was a 68:32 mixture of the 3R,4R and 3S,4S enantiomers. The phenanthrene trans-9,10-dihydrodiol produced by P. chrysosporium was predominantly the 9S,10S enantiomer, but those produced by C. elegans and S. racemosum were predominantly the 9R,10R enantiomer. The results indicate that although different fungi may exhibit similar regioselectivity, there still may be differences in stereoselectivity that depend on the species and the cultural conditions.
A set of 21 polyheteroaromatic compounds substituted with flexible cationic groups and of similar molecular size has been analyzed for binding with DNA and for effects of the bleomycin-mediated degradation of the DNA double helix. Increases in apparent rates of the DNA digestion were observed in all cases under the experimental conditions of noncompetitive binding of these compounds and bleomycin to DNA. Surprisingly, the quantitative structure-activity relationship analysis revealed two distinct correlations despite close structural similarities for the set of bleomycin amplifiers. These unusual results are explained in terms of the formation of two stereochemically different ternary complexes of activated bleomycin-DNA-amplifier. The relevance of this finding for the design of new bleomycin amplifiers is discussed.