London Plane Tree bioaerosol exposure and allergic sensitization in Sydney, Australia.

BACKGROUND: Exposure to London Plane Tree (Platanus) bioaerosols in Sydney, Australia has been anecdotally linked to respiratory irritation, rhinitis, and conjunctivitis. OBJECTIVE: To determine the relationships between Platanus bioaerosol exposure, allergic sensitization, and symptoms. METHODS: Sixty-four subjects with self-reported Platanus symptoms were recruited from inner-urban Sydney. Allergic sensitization was determined by skin prick test (SPT) to 13 allergens. Airborne concentrations of Platanus pollen, trichomes, and achene fibers, and other pollen and fungal spores, were measured over the spring and summer of 2006-2007. Subjects' allergic symptoms were monitored concurrently. The Halogen immunoassay (HIA) was used to measure subjects' immunoglobulin E (IgE) reactivity to collected bioaerosols. RESULTS: Platanus pollen constituted 76% of total pollen between July 2006 and April 2007. Airborne concentrations of Platanus pollen peaked from August until October. Non-Platanus pollen peaked from July to December. Elevated concentrations of trichomes and achene fibers occurred from September to December and August to October, respectively. As determined by SPT, 85.9% of subjects were sensitized, 65.6% to any pollen tested, 56.3% to Lolium perenne, and 23.4% to Platanus. Higher mean daily symptom scores were only associated with high counts of non-Platanus pollens. HIA analysis demonstrated IgE binding to Platanus pollen in all Platanus sensitized subjects. Personal nasal air sampling detected airborne trichomes that were capable of being inhaled. Platanus trichomes or achene fibers did not bind IgE from any subject. CONCLUSIONS: Platanus bioaerosols exist in high concentrations between August and November in inner-urban Sydney but were not associated with seasonal symptoms. Platanus trichomes are inhaled and may constitute a respiratory irritant. TRIAL REGISTRATION: Clinicaltrials.gov Identifier: NCTXXXXX.

The vertical distribution of house dust mite allergen in carpet and the effect of dry vacuum cleaning.

BACKGROUND: Carpets are large reservoirs of the dust mite allergen Der p 1. The effect of vacuum cleaning on the distribution of Der p 1 in carpets is poorly understood. METHODS: Samples were cut from 7 used household carpets, all over 5 years of age. From each carpet, 10 samples were left untreated, 10 vacuumed with an upright vacuum cleaner, and 10 vacuumed with a canister vacuum cleaner. Each section was then cut into 3 horizontal layers: the top 2mm, the remainder of the carpet pile, and the carpet base. The mass of Der p 1 as a proportion of carpet volume was then determined. RESULTS: The concentrations of Der p 1 in each depth layer varied considerably between the 7 untreated carpets. In the top layer, Der p 1 concentrations were (median; 25th-75th percentiles): 41.9; 28.3-92.6 pg/mm3. For the middle layer they were similar (38.1; 22.4-108.5 pg/mm3), and for the carpet base, higher (212.4; 98.8-456.2 pg/mm3). In most cases, cleaning using either type of vacuum cleaner resulted in no significant reduction in allergen concentration throughout all depth layers. A subset of carpets showed an apparent increase in Der p 1 concentration in one or more layers following vacuum cleaning. In all tests Der p 1 was collected in the vacuum cleaners' filter bags. CONCLUSIONS: The depth-distribution of Der p 1 differs widely amongst used carpets. Vacuum cleaning changes the distribution of Der p 1 within such carpets but does not necessarily result in a reduction in the overall content.

Detection of allergens from Alternaria alternata by gold-conjugated anti-human IgE and field emission scanning electron microscopy.

Fungal allergens are present in viable and non-viable conidia, hyphae and fungal fragments. It has been shown that large quantities of allergen are released from conidia during germination. We used a gold immunolabelling technique and field emission scanning electron microscopy to examine the allergen release from Alternaria alternata conidia. Immunolabelling was associated with the hyphal tip and amorphous matter associated with the emerging hyphae. Non-specific antibody controls showed no labelling associated with germinating fungi. This suggests that material released from hyphae may be an additional source of fungal allergens.

Airborne fungal fragments and allergenicity.

Exposure to fungi, particularly in water damaged indoor environments, has been thought to exacerbate a number of adverse health effects, ranging from subjective symptoms such as fatigue, cognitive difficulties or memory loss to more definable diseases such as allergy, asthma and hypersensitivity pneumonitis. Understanding the role of fungal exposure in these environments has been limited by methodological difficulties in enumerating and identifying various fungal components in environmental samples. Consequently, data on personal exposure and sensitization to fungal allergens are mainly based on the assessment of a few select and easily identifiable species. The contribution of other airborne spores, hyphae and fungal fragments to exposure and allergic sensitization are poorly characterized. There is increased interest in the role of aerosolized fungal fragments following reports that the combination of hyphal fragments and spore counts improved the association with asthma severity. These fragments are particles derived from any intracellular or extracellular fungal structure and are categorized as either submicron particles or larger fungal fragments. In vitro studies have shown that submicron particles of several fungal species are aerosolized in much higher concentrations (300-500 times) than spores, and that respiratory deposition models suggest that such fragments of Stachybotrys chartarum may be deposited in 230-250 fold higher numbers than spores. The practical implications of these models are yet to be clarified for human exposure assessments and clinical disease. We have developed innovative immunodetection techniques to determine the extent to which larger fungal fragments, including hyphae and fractured conidia, function as aeroallergen sources. These techniques were based on the Halogen Immunoassay (HIA), an immunostaining technique that detects antigens associated with individual airborne particles >1 microm, with human serum immunoglobulin E (IgE). Our studies demonstrated that the numbers of total airborne hyphae were often significantly higher in concentration than conidia of individual allergenic genera. Approximately 25% of all hyphal fragments expressed detectable allergen and the resultant localization of IgE immunostaining was heterogeneous among the hyphae. Furthermore, conidia of ten genera that were previously uncharacterized could be identified as sources of allergens. These findings highlight the contribution of larger fungal fragments as aeroallergen sources and present a new paradigm of fungal exposure. Direct evidence of the associations between fungal fragments and building-related disease is lacking and in order to gain a better understanding, it will be necessary to develop diagnostic reagents and detection methods, particularly for submicron particles. Assays using monoclonal antibodies enable the measurement of individual antigens but interpretation can be confounded by cross-reactivity between fungal species. The recent development of species-specific monoclonal antibodies, used in combination with a fluorescent-confocal HIA technique should, for the first time, enable the speciation of morphologically indiscernible fungal fragments. The application of this novel method will help to characterize the contribution of fungal fragments to adverse health effects due to fungi and provide patient-specific exposure and sensitization profiles.