Childhood fish oil supplementation modifies associations between traffic related air pollution and allergic sensitisation.

BACKGROUND: Studies of potential adverse effects of traffic related air pollution (TRAP) on allergic disease have had mixed findings. Nutritional studies to examine whether fish oil supplementation may protect against development of allergic disease through their anti-inflammatory actions have also had mixed findings. Extremely few studies to date have considered whether air pollution and dietary factors such as fish oil intake may interact, which was the rationale for this study. METHODS: We conducted a secondary analysis of the Childhood Asthma Prevention Study (CAPS) birth cohort, where children were randomised to fish oil supplementation or placebo from early life to age 5 years. We examined interactions between supplementation and TRAP (using weighted road density at place of residence as our measure of traffic related air pollution exposure) with allergic disease and lung function outcomes at age 5 and 8 years. RESULTS: Outcome information was available on approximately 400 children (~ 70% of the original birth cohort). Statistically significant interactions between fish oil supplementation and TRAP were seen for house dust mite (HDM), inhalant and all-allergen skin prick tests (SPTs) and for HDM-specific interleukin-5 response at age 5. Adjusting for relevant confounders, relative risks (RRs) for positive HDM SPT were RR 1.74 (95% CI 1.22-2.48) per 100 m local road or 33.3 m of motorway within 50 m of the home for those randomised to the control group and 1.03 (0.76-1.41) for those randomised to receive the fish oil supplement. The risk differential was highest in an analysis restricted to those who did not change address between ages 5 and 8 years. In this sub-group, supplementation also protected against the effect of traffic exposure on pre-bronchodilator FEV1/FVC ratio. CONCLUSIONS: Results suggest that fish oil supplementation may protect against pro-allergic sensitisation effects of TRAP exposure. Strengths of this analysis are that supplementation was randomised and independent of TRAP exposure, however, findings need to be confirmed in a larger experimental study with the interaction investigated as a primary hypothesis, potentially also exploring epigenetic mechanisms. More generally, studies of adverse health effects of air pollution may benefit from considering potential effect modification by diet and other factors. TRIAL REGISTRATION: Australia New Zealand Clinical Trial Registry. www.anzctr.org.au Registration: ACTRN12605000042640 , Date: 26th July 2005. Retrospectively registered, trial commenced prior to registry availability.

DETECTION OF AEROSOLIZED BACTERIA IN EXPIRED AIR SAMPLES FROM ASIAN ELEPHANTS (ELEPHAS MAXIMUS).

Elephant-mediated transmission of tuberculosis is assumed to be similar to human models, which state close and prolonged contact with an infected individual is required for transmission. Although considered a risk factor for infection, several case studies have reported that close contact with an elephant is not always necessary for transmission, and the role of aerosolized bacteria remains unclear. To investigate aerosol-mediated transmission of pathogenic bacteria from elephants, a method for the detection of aerosols using an adapted sampling system was developed. A commensal bacterium was isolated from the upper respiratory tract of elephants ( Elephas maximus ) and was used as a proxy organism to detect aerosolized droplets in the sampling system. It was found that elephants are capable of producing aerosolized bacterial particles of a size small enough to remain airborne for prolonged periods and penetrate the lower regions of the human respiratory tract.

Peanut protein extraction conditions strongly influence yield of allergens Ara h 1 and 2 and sensitivity of immunoassays.

The clinical importance of peanut (Arachis hypogaea) allergies demands standardized allergen extraction protocols. We determined the effectiveness of common extraction conditions (20 buffers, defatting reagents, extraction time/temperatures, processing, extraction repeats) on crude protein and Ara h 1 and 2 yields. Despite similar 1D-gel profiles, defatting with n-hexane resulted in significantly higher yields of crude protein, Ara h 1, and Ara h 2 than with diethyl ether. The yields were affected by the composition and pH of the extraction buffers and other conditions, but crude protein yield did not always correlate with Ara h 1 and 2 yields. Denaturants, reducing agents, acidic buffers, and thermal processing of peanuts perturbed allergen quantification in ELISAs, probably via exposure of additional epitopes. Allergen detection in 2D-Western blots with PBS resulted in greater sensitivity than with TBS or Tris. We recommend that allergen extraction conditions be selected based on the research question being investigated.

Time-Based Measurement of Personal Mite Allergen Bioaerosol Exposure over 24 Hour Periods.

Allergic diseases such as asthma and rhinitis are common in many countries. Globally the most common allergen associated with symptoms is produced by house dust mites. Although the bed has often been cited as the main site of exposure to mite allergens, surprisingly this has not yet been directly established by measurement due to a lack of suitable methods. Here we report on the development of novel methods to determine the pattern of personal exposure to mite allergen bioaerosols over 24-hour periods and applied this in a small field study using 10 normal adults. Air was sampled using a miniature time-based air-sampler of in-house design located close to the breathing zone of the participants, co-located with a miniature time-lapse camera. Airborne particles, drawn into the sampler at 2L/min via a narrow slot, were impacted onto the peripheral surface of a disk mounted on the hour-hand of either a 12 or 24 hour clock motor. The impaction surface was either an electret cloth, or an adhesive film; both novel for these purposes. Following a review of the time-lapse images, disks were post-hoc cut into subsamples corresponding to eight predetermined categories of indoor or outdoor location, extracted and analysed for mite allergen Der p 1 by an amplified ELISA. Allergen was detected in 57.2% of the total of 353 subsamples collected during 20 days of sampling. Exposure patterns varied over time. Higher concentrations of airborne mite allergen were typically measured in samples collected from domestic locations in the day and evening. Indoor domestic Der p 1 exposures accounted for 59.5% of total exposure, whereas total in-bed-asleep exposure, which varied 80 fold between individuals, accounted overall for 9.85% of total exposure, suggesting beds are not often the main site of exposure. This study establishes the feasibility of novel methods for determining the time-geography of personal exposure to many bioaerosols and identifies new areas for future technical development and clinical applications.

Absence of back to school peaks in human rhinovirus detections and respiratory symptoms in a cohort of children with asthma.

Much of what is known about the seasonality of human rhinovirus (hRV) infections has been learned from the study of acute asthma exacerbations presenting to emergency care, including those among children at the start of the school term. Much less is known about the patterns of hRVs in the community. In this study, viruses and day-to-day symptoms of asthma and colds were monitored twice weekly in 67 children with asthma aged 5-12 years, over a 15 month period in Sydney, Australia. Overall hRV was detected in 314/1232 (25.5%) of nasal wash samples and 142/1231 (11.5%) of exhaled breath samples; of these, 231 and 24 respectively were genotyped. HRVs were detected with similar prevalence rate throughout the year, including no peak in hRV prevalence following return to school. No peaks were seen in asthma and cold symptoms using twice-weekly diary records. However, over the same period in the community, there were peaks in asthma emergency visits both at a large local hospital and in state-wide hospitalizations, following both return to school (February) and in late autumn (May) in children of the same age. This study suggests that hRV infections are common throughout the year among children, and differences in virus prevalence alone may not account for peaks in asthma symptoms.

Rhinoviruses significantly affect day-to-day respiratory symptoms of children with asthma.

BACKGROUND: Viruses are frequently associated with acute exacerbations of asthma, but the extent to which they contribute to the level of day-to-day symptom control is less clear. OBJECTIVE: We sought to explore the relationship between viral infections, host and environmental factors, and respiratory symptoms in children. METHODS: Sixty-seven asthmatic children collected samples twice weekly for an average of 10 weeks. These included nasal wash fluid and exhaled breath for PCR-based detection of viral RNA, lung function measurements, and records of medication use and asthma and respiratory symptoms in the previous 3 days. Atopy, mite allergen exposure, and vitamin D levels were also measured. Mixed-model regression analyses were performed. RESULTS: Human rhinoviruses (hRVs) were detected in 25.5% of 1232 nasal samples and 11.5% of breath samples. Non-hRV viruses were detected in less than 3% of samples. hRV in nasal samples was associated with asthma symptoms (cough and phlegm: odds ratio = 2.0; 95% CI = 1.4-2.86, P = .0001; wheeze and chest tightness: odds ratio = 2.34, 95% CI = 1.55-3.52, P < .0001) and with cold symptoms, as reported concurrently with sampling and 3 to 4 days later. No differences were found between the 3 hRV genotypes (hRV-A, hRV-B, and hRV-C) in symptom risk. A history of inhaled corticosteroid use, but not atopic status, mite allergen exposure, or vitamin D levels, modified the association between viruses and asthma symptoms. CONCLUSION: The detection of nasal hRV was associated with a significantly increased risk of day-to-day asthma symptoms in children. Host, virus genotype, and environmental factors each had only a small or no effect on the relationship of viral infections to asthma symptoms.

Respiratory virus RNA is detectable in airborne and droplet particles.

Aerosol transmission routes of respiratory viruses have been classified by the WHO on the basis of equilibrium particle size. Droplet transmission is associated with particles sized >5 µm in diameter and airborne transmission is associated with particles sized ≤5 µm in diameter. Current infection control measures for respiratory viruses are directed at preventing droplet transmission, although epidemiological evidence suggests concurrent airborne transmission also occurs. Understanding the size of particles carrying viruses can be used to inform infection control procedures and therefore reduce virus transmission. This study determined the size of particles carrying respiratory viral RNA produced on coughing and breathing by 12 adults and 41 children with symptomatic respiratory infections. A modified six-stage Andersen Sampler collected expelled particles. Each stage was washed to recover samples for viral RNA extraction. Influenza A and B, parainfluenza 1, 2 and 3, respiratory syncytial virus (RSV), human metapneumovirus and human rhinoviruses (hRV) were detected using RT-PCR. On breathing, 58% of participants produced large particles (>5 µm) containing viral RNA and 80% produced small particles (≤5 µm) carrying viral RNA. On coughing, 57% of participants produced large particles containing viral RNA and 82% produced small particles containing viral RNA. Forty five percent of participants produced samples positive for hRV viral RNA and 26% of participants produced samples positive for viral RNA from parainfluenza viruses. This study demonstrates that individuals with symptomatic respiratory viral infections produce both large and small particles carrying viral RNA on coughing and breathing.

Most personal exposure to house dust mite aeroallergen occurs during the day.

BACKGROUND: The bed is commonly regarded as the main site of house dust mite exposure; however this has not been directly established by continuous measurements. The objective of this study was to determine the pattern of personal exposure to mite aeroallergen over 24 hours. METHODS: 12 adults each collected 9 sequential samples (8 during the day, mean 115 mins, and one overnight, mean 514 mins) over 24 hours using a portable air-pump (2L/min) connected to an IOM filter located on the shoulder during the day and on the bed head overnight. Samples were analysed for mite allergen Der p 1 by ELISA. Location and activity were recorded. A mixed model analysis was performed to determine exposure as a function of 14 categories of activity. RESULTS: Personal aeroallergen exposure differed widely over time, both within and between subjects. The highest average exposure (1117 pg/m(3), 95% CI: 289-4314) occurred on public transport and the lowest overnight in bed (45 pg/m(3), 95% CI: 17-17), which contributed only 9.8% (95% CI: 4.4%-15.1%) of total daily exposure. Aeroallergens were not related to bed reservoirs. CONCLUSION: The study challenges the current paradigm that the bed is the main site of HDM exposure and instead suggests most exposure occurs in association with domestic activity and proximity to other people. Effective mite interventions, designed to improve asthma outcomes, need to first identify and then address the multiple sources of aeroallergen exposure.

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.

It's time to rethink mite allergen avoidance.

The role of allergen exposure in the etiology of allergic sensitization and asthma is complex. Advice on strategies to avoid domestic allergens remains contentious because trials of interventions aimed to prevent asthma or reduce symptoms have often failed to demonstrate benefits. Asthma management guidelines differ widely in their recommendations, while Web-based information often claims benefits associated with products. In this rostrum we argue that although many factors have a role in both the etiology and the exacerbation of asthma, allergen exposure probably remains an important contributor to the manifestations of the disease. Currently, there is no evidence-based framework for effective domestic allergen avoidance interventions to reduce chronic aeroallergen exposure. The development of an effective approach to allergen avoidance requires a better understanding of (a) the physical nature of chronic aeroallergen exposure and methods for measuring and reducing this, (b) the interaction between allergen exposure and innate immune modulators at different disease stages, and (c) markers enabling the identification of individuals who would benefit from this. The strategic risk of overemphasizing other novel mechanisms and approaches to asthma management is that we will prematurely abandon and fail to improve an existing approach that could have a significant impact on the development, progression, and symptoms of the disease.

Effects of early cat or dog ownership on sensitisation and asthma in a high-risk cohort without disease-related modification of exposure.

Variation in the observed association between pet ownership and allergic disease may be attributable to selection bias and confounding. The aim of this study was to suggest a method to assess disease-related modification of exposure and second to examine how cat acquisition or dog ownership in early life affects atopy and asthma at 5 years. Information on sociodemographic factors and cat and dog ownership was collected longitudinally in an initially cat-free Australian birth cohort based on children with a family history of asthma. At age 5 years, 516 children were assessed for wheezing, and 488 for sensitisation. Data showed that by age 5 years, 82 children had acquired a cat. Early manifestations of allergic disease did not foreshadow a reduced rate of subsequent acquisition of a cat. Independent risk factors for acquiring a cat were exposure to tobacco smoke at home odds ratio (OR) 1.92 [95% confidence interval (CI) 1.13, 3.26], maternal education < or =12 years OR 1.95 [1.08, 3.51] and dog ownership OR 2.23 [1.23, 4.05]. Cat or dog exposure in the first 5 years was associated with a decreased risk of any allergen sensitisation, OR 0.50 [0.28, 0.88] but no association with wheeze OR 0.96 [0.57, 1.61]. This risk was not affected by age at which the cat was acquired or whether the pet was kept in- or outdoors. In conclusion, cat or dog ownership reduced the risk of subsequent atopy in this high-risk birth cohort. This cannot be explained by disease-related modification of exposure. Public health recommendations on the effect of cat and dog ownership should be based on birth cohort studies where possible selection bias has been taken into account.

Exhalation of respiratory viruses by breathing, coughing, and talking.

There is a lack of quantitative information about the generation of virus aerosols by infected subjects. The exhaled aerosols generated by coughing, talking, and breathing were sampled in 50 subjects using a novel mask, and analyzed using PCR for nine respiratory viruses. The exhaled samples from a subset of 10 subjects who were PCR positive for rhinovirus were also examined by cell culture for this virus. Of the 50 subjects, among the 33 with symptoms of upper respiratory tract infections, 21 had at least one virus detected by PCR, while amongst the 17 asymptomatic subjects, 4 had a virus detected by PCR. Overall, rhinovirus was detected in 19 subjects, influenza in 4 subjects, parainfluenza in 2 subjects, and human metapneumovirus in 1 subject. Two subjects were co-infected. Of the 25 subjects who had virus-positive nasal mucus, the same virus type was detected in 12 breathing samples, 8 talking samples, and in 2 coughing samples. In the subset of exhaled samples from 10 subjects examined by culture, infective rhinovirus was detected in 2. These data provide further evidence that breathing may be a source of respirable particles carrying infectious virus.

Surveillance of Fungal Allergic Sensitization Using the Fluorescent Halogen Immunoassay.

OBJECTIVE: Conidia derived from a small number of common fungal genera are widely accepted as the etiological agents responsible for fungal allergic sensitization. The contribution of fungal conidia, spores, airborne hyphae, and subcellular fragments from other uncharacterized fungal genera remains unclear. In this proof-of-concept study, we examined the composition of mycoaerosols that atopic women were exposed and sensitized to in their own indoor environment using the fluorescent halogen immunoassay (fHIA). PATIENTS AND METHODS: Mycoaerosols were collected onto mixed cellulose ester protein binding membranes (PBMs) for 30 minutes with volumetric air sampling pumps. The PBMs were laminated with an adhesive cover slip and indirectly immunostained with individual patient serum IgE using the fHIA. Samples were examined using confocal laser scanning microscopy and immunostained particles were expressed as a percentage of total particles. RESULTS: All air samples contained a broad spectrum of fungal spores, conidia, hyphae, and other fungal particulates. Airborne concentrations varied between individual study participant environments. Positively immunostained conidia belonging to moniliaceous amerospores, Cladosporium, Alternaria, and many unknown species were observed in the majority of air samples. Other fungal genera including Bipolaris, Curvularia, Pithomyces, and Stachybotrys, in addition to, ascospore genera and dematiaceous hyphal fragments released detectable allergen. Twelve percent of all fHIA haloes quantified in the analysis were directed towards fungal particles. No immunostaining was detected to conidia belonging to Epicoccum, Fusarium, and Spegazzinia species. CONCLUSION: In addition to characterized fungal aeroallergens, we observed a wider composition of fungi that bound human IgE. Field surveillance studies that utilize immunodiagnostic techniques such as the fHIA will provide further insight into the diversity of fungi that function as aeroallergen sources in individual study participant environments.

Nonlinear relationship of mite allergen exposure to mite sensitization and asthma in a birth cohort.

BACKGROUND: There is uncertainty about the nature of the relationship between mite allergen exposure during infancy and the expression of allergic diseases in childhood. OBJECTIVE: We sought to explore the relationships between repeated measurements of mite allergen exposure during the first 5 years of life and clinical allergic disease outcomes at age 5 years. METHODS: In a birth cohort of 516 at-risk children, 13 bed dust samples were collected between birth and 5 years of age and analyzed for mite allergen. At age 5 years, the presence of mite atopy was assessed based on skin prick test results, and clinical assessments for asthma, eczema, and wheeze were conducted. The association of allergen exposure with each clinical outcome was examined by means of logistic regression, with adjustments for potential confounders. RESULTS: The lowest and highest mite exposure quintiles, expressed as mean allergen concentration averaged over 0 to 5 years, were associated with a lower prevalence of mite atopy and of asthma compared with intermediate levels of exposure. These relationships, when determined by using average allergen concentration over 0 to 18 months, were weaker. CONCLUSIONS: This study demonstrates a nonlinear relationship between mite allergen exposure and clinical outcomes in this generally high mite allergen environment.

Allergen avoidance.

The systematic avoidance of indoor allergens by modification of houses, furnishings, or hygiene practices has long been advocated to reduce both the incidence of allergic diseases in at-risk infants and exacerbations or symptoms in those previously sensitized with such a disease. However, such advocacy is now under challenge, due to both a lack of evidence of clinical efficacy of avoidance measures and rapidly changing models of disease causation and pathology. This article reviews the current dilemmas in understanding the complexity of allergen sources, disease risks, nature of bioaerosol exposure, and the inadequacies of many available methods.

A new method for sampling and detection of exhaled respiratory virus aerosols.

We have developed a mask sampler for exhaled respiratory viruses. Among a group of 9 patients with cold symptoms who had virus-positive nasal mucus specimens, as analyzed by multiplexed polymerase chain reaction, virus-positive mask samples were obtained after coughing (20 times), talking (20 min), or breathing (20 min) from 6, 5, and 3 patients, respectively.