Measurement of Horse Allergens Equ c 1 and Equ c 2: A Comparison among Breeds.

INTRODUCTION: Horse allergens are less studied than allergens from other furry animals and these allergens must be evaluated to understand the complexity of allergy to horses. The aims of this study were to develop assays for the horse allergens Equ c 1 and Equ c 2 in dander and saliva and to determine their levels in ten horse breeds. The study also included a comparison of these findings with previous results on the levels of Equ c 4 performed on the same study population. METHOD: The study population included 170 horses from 10 horse breeds including American Curly and Russian Bashkir horse, which have been suggested to be hypoallergenic. Competitive ELISA assays were developed, with polyclonal antibodies as capture antibodies, for the detection of Equ c 1 and Equ c 2 in dander and saliva samples. RESULTS: The horse allergens Equ c 1 and Equ c 2 were found in all dander and saliva samples from the ten horse breeds. The GM level (ng/µg protein) of Equ c 1 in dander was 470 (range 129-2,569) and in saliva samples, 40 (range 6-160). The GM level of Equ c 2 in dander was 138 (range 18-1,650) and in saliva samples, 0.8 (range 0.03-17). In dander, there were no significant differences in Equ c 1 and Equ c 2 GM levels between stallions, mares, and geldings. CONCLUSION: Our results show high intra- and inter-breed variability. Neither the American Curly horse nor the Russian Bashkir horse, earlier categorized as hypoallergenic breeds, was associated with lower allergen levels of Equ c 1, Equ c 2, or Equ c 4 than the other horse breeds investigated.

Levels of horse allergen Equ c 4 in dander and saliva from ten horse breeds.

BACKGROUND: Horses are an important source of allergens, but the distribution of horse allergens is poorly understood. Five horse allergens have been identified, Equ c 1-4 and 6. Equ c 4 seems to be an important allergen, with an IgE-binding frequency of 77% in horse-sensitized individuals. OBJECTIVES: The aim of this study was to investigate levels of horse allergen Equ c 4 in dander, saliva and urine from ten horse breeds. METHOD: The study population included 170 horses (87 mares, 27 stallions, 56 geldings) from ten breeds. Horse dander, saliva and urine samples were collected. Levels of horse allergen Equ c 4 were quantified using a two-site sandwich ELISA (mAb 103 and 14G4) and were expressed as Equ c 4 U/µg protein. RESULTS: The horse allergen Equ c 4 was present in all dander and saliva samples from ten horse breeds, with high within-breed and inter-breed variations; GM values were 639 Equ c 4 U/µg protein (range 5-15 264) for dander and 39.5 (4-263) for saliva. Equ c 4 was found in 19/21 urine samples. Adjusted for age, sex and changes over time, no differences between breeds could be seen in dander, while in saliva the North Swedish horse showed lower levels of Equ c 4 than any other breed. The levels of Equ c 4 protein in dander and saliva were significantly higher in samples from stallions compared to mares and geldings, independent of breed. CONCLUSIONS AND CLINICAL RELEVANCE: The results show a high variability in allergen levels of Equ c 4 in dander and saliva both within and between breeds. Significantly higher levels were found in stallions compared to mares and geldings, independent of breed. Results suggest that none of the horse breeds studied can be recommended for individuals allergic to Equ c 4.

Use of a 3-D dispersion model for calculation of distribution of horse allergen and odor around horse facilities.

The interest in equestrian sports has increased substantially during the last decades, resulting in increased number of horse facilities around urban areas. In Sweden, new guidelines for safe distance have been decided based on the size of the horse facility (e.g., number of horses) and local conditions, such as topography and meteorology. There is therefore an increasing need to estimate dispersion of horse allergens to be used, for example, in the planning processes for new residential areas in the vicinity of horse facilities. The aim of this study was to develop a method for calculating short- and long-term emissions and dispersion of horse allergen and odor around horse facilities. First, a method was developed to estimate horse allergen and odor emissions at hourly resolution based on field measurements. Secondly, these emission factors were used to calculate concentrations of horse allergen and odor by using 3-D dispersion modeling. Results from these calculations showed that horse allergens spread up to about 200 m, after which concentration levels were very low (<2 U/m³). Approximately 10% of a study-group detected the smell of manure at 60m, while the majority--80%-90%--detected smell at 60 m or shorter distance from the manure heap. Modeling enabled horse allergen exposure concentrations to be determined with good time resolution.

Systematic review of respiratory health among dairy workers.

The dairy industry is changing on a global scale with larger, more efficient operations. The impact of this change on worker health and safety, specifically, associations between occupational lung disease and inhalation exposures, has yet to be reported in a comprehensive review of the scientific literature. Therefore, a three-tier process was used to identify information using a keyword search of online databases of scientific literature. Of the 147 citations reviewed, 52 met initial screening criteria, and 30 were included in this review. Dairy workers experience lung conditions such as asthma, chronic obstructive pulmonary disease, hypersensitivity pneumonitis, chronic bronchitis, and cancer. Recent pulmonary function studies have identified obstructive lung changes among dairy farm workers. The increased scale of dairy production with significant changes in technology and work practices has altered inhalation exposure patterns among dairy workers. The inhalation exposure in the dairy work environment may elicit differing inflammatory responses in relation to timing of initial exposure as well as to repeated exposures. Few studies have measured inhalation exposure while simultaneously assessing the impact of the exposure on lung function of dairy farm workers. Even fewer studies have been implemented to assess the impact of aerosol control technology to reduce inhalation exposure. Future research should evaluate worker exposure to aerosols through a task-based approach while utilizing novel methods to assess inhalation exposure and associated inflammatory responses. Finally, potential solutions should be developed and tested to reduce inhalation exposure to inflammatory agents and respiratory diseases in the dairy farm work environment.

Installation of mechanical ventilation in a horse stable: effects on air quality and human and equine airways.

OBJECTIVES: To examine the effects of installing a mechanical ventilation system at a riding-school stable on indoor air quality and human and horse airways. METHODS: The intervention was the installation of mechanical ventilation in a riding-school stable. Carbon dioxide (CO2), ammonia, particles, horse allergen, microorganisms and endotoxins were measured in the stable. The stable-workers and riding-students completed a questionnaire and underwent the following tests: analysis of nasal lavage for inflammation biomarkers; levels of exhaled nitrogen oxide (NO); measurements of daily peak-expiratory flow (PEF). The horses were examined clinically by airway endoscopy and bronchoalveolar lavage (BAL) and were analysed for cytology and biomarkers. RESULTS: Levels of CO2 were nearly halved and airborne horse allergen levels were markedly reduced (5-0.8 kU/m3) after the intervention. A decreased level of ultrafine particles was observed (8000-5400 particles/cm3) after the intervention, while total and respirable dust levels were mainly unchanged (200 and 130 µg/m3). Levels of microorganisms in surface samples decreased following the intervention, whereas airborne microorganisms and endotoxin increased. There was no significant change in human symptoms, PEF-variability, exhaled NO or inflammatory biomarkers in the nasal lavage. In horses, the mean score of lower airway mucus was significantly reduced together with the mean level of expression of interleukin-6 mRNA in BAL cells after the intervention. CONCLUSIONS: The installation of a mechanical ventilation system resulted in an increased air exchange rate, as demonstrated by reduced levels of CO2, ammonia, ultrafine particles and horse allergen. There was no significant clinical effect on human airways, but there was a tendency for reduced inflammation markers. The results on the horses may indicate less impact on their airways after the intervention.

Respiratory health among Korean pupils in relation to home, school and outdoor environment.

There are few studies about school-environment in relation to pupils' respiratory health, and Korean school-environment has not been characterized. All pupils in 4th grade in 12 selected schools in three urban cities in Korea received a questionnaire (n = 2,453), 96% participated. Gaseous pollutants and ultrafine particles (UFPs) were measured indoors (n = 34) and outdoors (n = 12) during winter, 2004. Indoor dampness at home was investigated by the questionnaire. To evaluate associations between respiratory health and environment, multiple logistic- and multi-level regression models were applied adjusting for potential confounders. The mean age of pupils was 10 yr and 49% were boys. No school had mechanical ventilation and CO(2)-levels exceeded 1,000 ppm in all except one of the classrooms. The indoor mean concentrations of SO(2), NO(2), O(3) and formaldehyde were 0.6 µg/m(3), 19 µg/m(3), 8 µg/m(3) and 28 µg/m(3), respectively. The average level of UFPs was 18,230 pt/cm(3) in the classrooms and 16,480 pt/cm(3) outdoors. There were positive associations between wheeze and outdoor NO(2), and between current asthma and outdoor UFPs. With dampness at home, pupils had more wheeze. In conclusion, outdoor UFPs and even low levels of NO(2) may adversely contribute to respiratory health in children. High CO(2)-levels in classrooms and indoor dampness/mold at home should be reduced.

Measurement of Horse Allergen (Equ cx) in Schools.

Background. The presence of horse allergen in public places is not well-known, unlike for instance cat and dog allergens, which have been studied extensively. The aim was to investigate the presence of horse allergen in schools and to what extent the influence of number of children with regular horse contact have on indoor allergen levels. Methods. Petri dishes were used to collect airborne dust samples during one week in classrooms. In some cases, vacuumed dust samples were also collected. All samples were extracted, frozen and analysed for Equ cx content shortly after sampling, and some were re-analysed six years later with a more sensitive ELISA assay. Results. Horse allergen levels were significantly higher in classrooms, in which many children had horse contact, regardless of sampling method. Allergen levels in extracts from Petri dish samples, which had been kept frozen, dropped about 53% over a six-year period. Conclusion. Horse allergen was present in classrooms and levels were higher in classrooms where many children had regular horse contact in their leisure time. This suggests that transfer of allergens takes place via contaminated clothing. Measures should be taken to minimize possible transfer and deposition of allergens in pet-free environments, such as schools.

Influence of horse stable environment on human airways.

BACKGROUND: Many people spend considerable amount of time each day in equine stable environments either as employees in the care and training of horses or in leisure activity. However, there are few studies available on how the stable environment affects human airways. This study examined in one horse stable qualitative differences in indoor air during winter and late summer conditions and assessed whether air quality was associated with clinically detectable respiratory signs or alterations to selected biomarkers of inflammation and lung function in stable personnel. METHODS: The horse stable environment and stable-workers (n = 13) in one stable were investigated three times; first in the winter, second in the interjacent late summer and the third time in the following winter stabling period. The stable measurements included levels of ammonia, hydrogen sulphide, total and respirable dust, airborne horse allergen, microorganisms, endotoxin and glucan. The stable-workers completed a questionnaire on respiratory symptoms, underwent nasal lavage with subsequent analysis of inflammation markers, and performed repeated measurements of pulmonary function. RESULTS: Measurements in the horse stable showed low organic dust levels and high horse allergen levels. Increased viable level of fungi in the air indicated a growing source in the stable. Air particle load as well as 1,3-beta-glucan was higher at the two winter time-points, whereas endotoxin levels were higher at the summer time-point. Two stable-workers showed signs of bronchial obstruction with increased PEF-variability, increased inflammation biomarkers relating to reported allergy, cold or smoking and reported partly work-related symptoms. Furthermore, two other stable-workers reported work-related airway symptoms, of which one had doctor's diagnosed asthma which was well treated. CONCLUSION: Biomarkers involved in the development of airway diseases have been studied in relation to environmental exposure levels in equine stables. Respirable dust and 1,3-beta-glucan levels were increased at winter stabling conditions. Some employees (3/13) had signs of bronchial obstruction, which may be aggravated by working in the stable environment. This study contributes to the identification of suitable biomarkers to monitor the indoor horse stable environment and the personnel. An improved management of the stable climate will be beneficial for the health of both stable workers and horses.

Acute health effects on planters of conifer seedlings treated with insecticides.

OBJECTIVES: The aim of this study was to assess acute health effects on planters caused by planting conifer seedlings treated with two insecticides, with active ingredients imidacloprid and cypermethrin, in comparison with untreated seedlings. METHODS: The investigation was a double-blind crossover study, which included a follow-up of 19 planters over a 3-week period. During Week 1, the 19 planters handled untreated conifer seedlings while they planted imidacloprid- and cypermethrin-treated seedlings during study Week 2 and 3, respectively. Signs and symptoms of acute health effects were documented by a questionnaire, administered by the field staff, during these 3 weeks. Inflammation markers in the nasal mucous membrane were also measured as an objective test. Exposure to cypermethrin was further assessed by measuring 3-phenoxybenzoic acid (3-PBA) in urine. No validated biomarker was available to assess internal exposure to imidacloprid. RESULTS: No clear, acute adverse health effects could be found in planters during the week of exposure to conifer seedlings treated with imidacloprid (Merit Forest) or cypermethrin (Forester), as compared to during the week of planting untreated seedlings. During the week of cypermethrin exposure, the individuals had 3-PBA values that were 12-54% higher (P < 0.05), depending on the worker, than those observed during the untreated week. There were no statistically significant correlations between the raised levels of 3-PBA and self-reported health problems. These results have been obtained during planting in late summer/early autumn and with good use of protective clothing. CONCLUSIONS: No clear, acute adverse health effects could be found in planters after exposure to conifer seedlings treated with imidacloprid (Merit Forest) or cypermethrin (Forester), as compared with planting untreated seedlings. The metabolite, 3-PBA, was found in low levels in urine and was increased after exposure to cypermethrin. However, no clear relationships could be found between exposure and reported symptoms or between elevated 3-PBA levels and reported symptoms.

Markers of respiratory inflammation in horses in relation to seasonal changes in air quality in a conventional racing stable.

Airborne factors in a conventionally managed racing stable and markers of pulmonary inflammation in the stabled horses were investigated on 3 occasions at 6-month intervals, including 2 winter periods and the intervening summer period. The stable measurements included inside and outside ambient temperature and relative humidity, levels of total and respirable dust, endotoxin, and 1,3-beta-glucan. Horses (n = 12) were examined in detail clinically as well as by endoscopy, bronchoalveolar lavage (BAL) cytology, and real-time polymerase chain reaction (RT-PCR) analysis of BAL-cells for IL-6 and IL-10 mRNA. Indoor measurements showed low dust levels irrespective of season. Inhalable dust, as well as 1,3-beta-glucan, were higher during the winter stabling period, whereas endotoxin levels were higher during summer. Complete data from all sampling occasions to be used for further evaluation was obtained for only 8 of the horses. There was a trend for elevation of BAL neutrophils in the horses during winter stabling that coincided with a 3.7-fold increased expression of IL-6 mRNA in BAL cells (P = 0.014). Compared to summer sampling, IL-10 mRNA expression was significantly upregulated in only 1 of the winter sampling occasions, implicating influence on immune regulation by factor/s apart from seasonal differences. Our findings suggest up-regulation of innate immunity in the airways of stabled horses; in particular involving IL-6 in association with mild elevations in respirable dust, 1,3-beta-glucan, and/or cold ambient air. However, given that this study was observational, other unmeasured environmental factors associated with winter stabling need to be considered.

Detection of horse allergen around a stable.

BACKGROUND: Integrating horse stables with built-up areas may lead to conflicts. Dispersion of horse allergen may become a health risk for allergic people. The aim was to measure the dispersion of horse allergen around a stable, considering wind speed and direction and vegetation. The disturbance of staff at a workplace nearby a stable was investigated. METHODS: Air sampling was performed around a stable (32 horses) at distances of 50-500 m in all directions. Sampling was done with a pump and an IOM sampler. Samples were collected at 50 points during all seasons. Horse allergen levels were determined using ELISA. Disturbance by horses was studied with a questionnaire handed to the employees in an office near the stable. RESULTS: The median horse allergen level at the stable entrance was 316 U/m(3), in the horse fields 40 U/m(3) and in the whole source area 16 U/m(3), which declined to <2 U/m(3) at about 50 m from the source area. Downwind of the prevailing winds low levels of horse allergen (2-4 U/m(3)) could sometimes be detected at up to 500 m. The staff, including those allergic to horses, managed to tolerate horses close to the workplace. CONCLUSIONS: At low winds horse allergen spread in ambient air about 50 m from the stable and horse fields. At higher winds low allergen levels were sometimes found in open areas up to 500 m from the source area. These levels were similar to those found in the office after moving away from the stable area. The employees did not report more symptoms of allergy or asthma while working close to the stable compared to after the move.

Different IgE reactivity profiles in birch pollen-sensitive patients from six European populations revealed by recombinant allergens: an imprint of local sensitization.

BACKGROUND: Sensitivity to birch pollen allergens is a common feature among European patients with seasonal pollen allergy. In this in vitro study, we examined the specific serum IgE binding profiles to individual birch pollen allergens in birch-sensitive patients from six European populations. METHODS: The study included 242 patients from Finland, Sweden, Austria, France, Switzerland and Italy. All suffered from seasonal rhinoconjunctivitis and/or asthma. Their sera were analyzed for specific IgE reactivity to individual birch pollen allergens (recombinant Bet v 1, Bet v 2 and Bet v 4) and natural birch pollen extract using Pharmacia CAP System and immunoblotting. RESULTS: Almost all Finnish, Swedish and Austrian sera contained IgE specific for Bet v 1 (>or=98%). Bet v 1-specific IgE antibodies were found in 90% of the French sera, and in 65 and 62% of the sera from Switzerland and Italy, respectively. Few Finnish (2%) and Swedish (12%) patients had IgE to Bet v 2, while Bet v 2 reactivity was more common in the other populations (20-43%). Reactivity to Bet v 4 was rare in all populations (5-11%) except for the Italian patients, in whom 3 of 11 sera were positive (27%). The immunoblot results supported the specific IgE profiles obtained with Pharmacia CAP System showing a broader IgE reactivity profile in patients from central and southern Europe as compared to northern Europe. CONCLUSION: Component-resolved allergy diagnosis with recombinant allergens reveals that the IgE reactivity profiles to individual birch pollen allergens vary between European populations. This observation may be explained by sensitization to different allergen sources and will have an impact on allergen-specific prevention and therapy strategies.