Development of a Sandwich ELISA to Measure Exposure to Occupational Cow Hair Allergens

Background: Cow hair and dander are important inducers of occupational allergies in cattle-exposed farmers. To estimate allergen exposure in farming environments, a sensitive enzyme immunoassay was developed to measure cow hair allergens. Methods: A sandwich ELISA was developed using polyclonal rabbit antibodies against a mixture of hair extracts from different cattle breeds. To assess the specificity of the assay, extracts from other mammalian epithelia, mites, molds and grains were tested. To validate the new assay, cow hair allergens were measured in passive airborne dust samples from the stables and homes of farmers. Dust was collected with electrostatic dust fall collectors (EDCs). Results: The sandwich ELISA was found to be very sensitive (detection limit: 0.1 ng/ml) and highly reproducible, demonstrating intra-and interassay coefficients of variation of 4 and 10%, respectively. The assay showed no reactivity with mites, molds and grains, but some cross-reactivity with other mammalian epithelia, with the strongest reaction with goat. Using EDCs for dust sampling, high concentrations of bovine allergens were measured in cow stables (4,760-559,400 mu g/m(2)). In addition, bovine allergens were detected in all areas of cattle farmer dwellings. A large variation was found between individual samples (0.3-900 mu g/m(2)) and significantly higher values were discovered in changing rooms. Conclusion: The ELISA developed for the detection of cow hair proteins is a useful tool for allergen quantification in occupational and home environments. Based on its low detection limit, this test is sensitive enough to detect allergens in passive airborne dust. Copyright (C) 2011 S. Karger AG, Base

Respiratory Allergens from Furred Mammals: Environmental and Occupational Exposure

Furry mammals kept as pets, farm and laboratory animals are important allergen sources. The prevalence of sensitization to furred mammals appears to be increasing worldwide. Several mammalian allergens from diverse species are well characterized with regard to their molecular structure and immunogenicity, and some are already available for component-resolved allergy diagnostics. The distribution of various mammalian allergens has been extensively studied during the past few decades. Animal allergens were found to be ubiquitous in the human environment, even in places where no animals reside, with concentrations differing considerably between locations and geographical regions. This review presents an overview of identified mammalian respiratory allergens classified according to protein families, and compiles the results of allergen exposure assessment studies conducted in different public and occupational environments

Respiratory allergens from furred mammals

Furry mammals kept as pets, farm and laboratory animals are important allergen sources. The prevalence of sensitization to furred mammals appears to be increasing worldwide. Several mammalian allergens from diverse species are well characterized with regard to their molecular structure and immunogenicity, and some are already available for component-resolved allergy diagnostics. The distribution of various mammalian allergens has been extensively studied during the past few decades. Animal allergens were found to be ubiquitous in the human environment, even in places where no animals reside, with concentrations differing considerably between locations and geographical regions. This review presents an overview of identified mammalian respiratory allergens classified according to protein families, and compiles the results of allergen exposure assessment studies conducted in different public and occupational environments

Animal allergens and their presence in the environment

Exposure to animal allergens is a major risk factor for sensitization and allergic diseases. Besides mites and cockroaches, the most important animal allergens are derived from mammals. Cat and dog allergies affect the general population; whereas, allergies to rodents or cattle is an occupational problem. Exposure to animal allergens is not limited to direct contact to animals. Based on their aerodynamic properties, mammalian allergens easily become airborne, attach to clothing and hair, and can be spread from one environment to another. For example, the major cat allergen Fel d 1 was frequently found in homes without pets and in public buildings, including schools, day-care centers, and hospitals. Allergen concentrations in a particular environment showed high variability depending on numerous factors. Assessment of allergen exposure levels is a stepwise process that involves dust collection, allergen quantification, and data analysis. Whereas a number of different dust sampling strategies are used, ELISA assays have prevailed in the last years as the standard technique for quantification of allergen concentrations. This review focuses on allergens arising from domestic, farm, and laboratory animals and describes the ubiquity of mammalian allergens in the human environment. It includes an overview of exposure assessment studies carried out in different indoor settings (homes, schools, workplaces) using numerous sampling and analytical methods and summarizes significant factors influencing exposure levels. However, methodological differences among studies have contributed to the variability of the findings and make comparisons between studies difficult. Therefore, a general standardization of methods is needed and recommended

Use of nasal filters for allergen exposure measurements in veterinary practices

Objectives: In this study, we applied novel nasal filters to assess animal allergen exposure of veterinary staff during their normal daily routine. Methods: Rhinix nasal filters were worn during work by 94 employees at different veterinary practices and 18 employees at a research institute, who acted as controls representing an animal-free environment. Contact with animals and the activities performed were documented by the study participants using a short questionnaire. Major allergens of cats (Fel d 1), dogs (Can f 1), and domestic mites (DM) were measured using fluorescence enzyme immunoassays. Results: At the practices, Can f 1 was detected in 98%, Fel d 1 in 82%, and DM allergens in 39% of the samples. Allergens were also detected in some control samples (6% for Can f 1, 39% for Fel d 1, and 17% for DM) but in very low concentrations. There was a highly significant difference between allergen levels in veterinary workers who treated at least one cat or dog during the sampling period and those who did not (2.66 vs. 0.70 ng/filter for Can f 1 and 1.01 vs. 0.20 ng/filter for Fel d 1). The amount of sampled Fel d 1 increased significantly with increasing duration of contact with cats. This effect was not observed for Can f 1. Conclusions: The majority of veterinary workers are exposed to dog and cat allergens, even without direct contact with these animals. Rhinix nasal filters may be considered a simple and easily applicable method for monitoring personal allergen exposure

Comparing the concentration levels of allergens and endotoxins in employees’ homes and offices

Objective!#!The aim of the study was to find out whether allergen and endotoxin concentrations in offices differ from those measured at the homes of employees, and identify the parameters that influence exposure.!##!Methods!#!Electrostatic dust collectors (EDCs) were placed in five office buildings (68 rooms, 436 EDCs), as well as the homes of the office workers (145 rooms, 405 EDCs) for 14 days, four times a year. In addition, surface samples were collected from the offices four times a year by vacuuming the carpeted floors. Domestic mite (DM), and the major cat and dog allergens (Fel d 1 and Can f 1) were quantified in all samples using fluorescence enzyme immunoassays. Endotoxin was measured in the EDC samples, using the Limulus amoebocyte lysate assay. The allergen and endotoxin concentrations were log transformed and analysed with multilevel models.!##!Results!#!Endotoxin concentrations were significantly higher in personal homes compared to levels measured in the offices, and depended on the number of persons living in each household, as well as the presence of a dog. DM allergens were significantly higher in households than in offices, and were significantly higher in bedrooms compared to living rooms. Offices occupied by cat owners had significantly higher Fel d 1 concentrations than offices or homes without. Additionally, Can f 1 concentrations were significantly higher in offices occupied by dog owners compared to those without.!##!Conclusions!#!Pet owners appear to transfer cat and dog allergens to their offices. Therefore, in case of allergy complaints at the office, employers and physicians might consider possible contamination by cat and dog allergens

Lower allergen levels in hypoallergenic Curly Horses? A comparison among breeds by measurements of horse allergens in hair and air samples.

BackgroundExposure to horses can cause severe allergic reactions in sensitized individuals. The breed, American Bashkir Curly Horse is categorized as hypoallergenic, primarily due to reports of allergic patients experiencing fewer symptoms while handling this special breed. The possible reasons for this phenomenon could be lower allergen production and/or reduced allergen release into the air because of increased sebum content in their skin and hair compared to other breeds. Therefore, the aim of the current study was to compare different horse breeds in relation to allergen content in hair and airborne dust samples.MethodsIn total, 224 hair samples from 32 different horse breeds were investigated. Personal nasal filters were used to collect airborne dust during the grooming of 20 Curly Horses and 20 Quarter Horses. Quantitative analysis of all samples was performed using two newly developed immunoassays for the detection of horse dander (HD) antigens and the major allergen Equ c 1 and the commercial assay for Equ c 4. Results were analyzed using multiple linear regression models for hair samples and the Mann Whitney U test for airborne samples.ResultsHorse antigen and allergen levels differed up to four orders of magnitude between individual animals. Despite enormous variability, levels of HD antigen, Equ c 1 and Equ c 4 in hair were significantly related to the breed and gender combined with the castration status of male animals. Curly Horses had significantly higher concentrations of all three tested parameters compared to the majority of the investigated breeds (medians: 11800 μg/g for HD antigen, 2400 μg/g for Equ c 1, and 258 kU/g for Equ c 4). Tinker Horses, Icelandic Horses and Shetland Ponies were associated with approximately 7-fold reduced levels of HD antigen and Equ c 1, and up to 25-fold reduced levels of Equ c 4 compared to Curly Horses. Compared to mares, stallions displayed increased concentrations of HD antigens, Equ c 1 and Equ c 4 by a factor 2.2, 3.5 and 6.7, respectively. No difference was observed between mares and geldings. No differences in airborne allergen concentrations collected with personal nasal filters during grooming were found between Curly and Quarter Horses.ConclusionBreed and castration status had a significant influence on the antigen and allergen levels of horse hair. However, these differences were smaller than the wide variability observed among individual horses. Compared to other breeds, Curly Horses were not associated with lower allergen levels in hair and in air samples collected during grooming. Our approach provides no molecular explanation why Curly Horses are considered to be hypoallergenic

Allergen levels in the hair of different cattle breeds

$\textit {Background:}$ Cattle are well-known sources of respiratory allergens in agricultural environments. Breed-specific differences in Bos d 2 (a major bovine allergen) levels in cattle hair have been previously suggested but not fully characterized. Therefore, the aim of the current study was to determine whether hair from common cattle breeds differs in protein and allergen content. $\textit {Methods:}$ In total, 80 hair samples from 16 different cattle breeds were analyzed. The protein concentration was determined using the Bradford assay. The allergen content was measured using a sandwich ELISA based on polyclonal antibodies against a bovine hair protein extract and a commercial immunoassay based on monoclonal antibodies against Bos d 2. Results are given in micrograms per gram of hair. Statistical analysis was performed using the Kruskal-Wallis test and Spearman’s rank correlation. $\textit {Results:}$ A wide variability in all 3 tested parameters was observed between the individual samples. The protein content differed by about 35-fold (0.3–12 mg/g), the bovine hair allergen content differed by about 500-fold (37–18,553 $\mu$g/g), and the Bos d 2 content differed by about 1,200-fold (5–6,323 $\mu$g/g). Protein, bovine hair allergen, and Bod d 2 values correlated strongly and significantly with one another. The median Bos d 2/bovine hair allergen ratio was 0.28. No significant differences were found between the most common breeds in Germany (Simmental, Holstein, and Braunvieh) and a group of rare breeds or between female and male animals. $\textit {Conclusions:}$ The results confirm a high variability in allergen levels between individual animals but also indicate that allergen production is related neither to the breed nor to gender