ABSTRACT
NABARUN GHOSH,1 GRISELDA ESTRADA,1 MITSY VELOZ,1 DANIUS BOUYI,1 JON BENNERT,2 JEFF BENNERT,2 CONSTANTINE SAADEH,3 and CHANDINI REVANNA4
1Life, Earth and Environmental Sciences, West Texas A&M University, Canyon, Texas 79015, USA, E-mail: nghosh@wtamu.edu, velozfam6401@yahoo.com; gestrada1@buffs.wtamu.edu, emersonbouyi@hotmail.com
2Air Oasis, Research and Development, Amarillo, Texas 79118, USA, E-mail: jon@airoasis.com; drj@airoasis.com
3Allergy A.R.T.S., Amarillo, Texas 79124, USA, E-mail: csaadeh@ allergyarts.com
4Department of Environmental Health and Safety, Texas Tech University, Lubbock, TX 79409, E-mail: c.revanna@ttu.edu
Abstract ................................................................................................. 102 4.1 Introduction .................................................................................. 103 4.2 Aeroallergens and Allergic Rhinitis ............................................. 106 4.3 Fungal Spores as Aeroallergens ................................................... 106 4.4 Fluorescence and Scanning Electron Microscopy on Pollen ....... 108 4.5 Installing and Preparing the Burkard Volumetric Spore Trap
for Pollen and Spore Analysis ...................................................... 108
4.6 Digital Microscopic Analysis of Collected Aeroallergens ............111 4.7 Observation on Pollen and Spores ................................................112 4.8 Effect of Meteorological Factors
on Distribution of Pollen and Spores ............................................112 4.9 Observation on Pollen with Fluorescence and Scanning
Electron Microscopy .....................................................................117 4.10 Concluding Remarks .................................................................. 121 Acknowledgements ............................................................................... 123 Keywords .............................................................................................. 123 References ............................................................................................. 124
ABSTRACT
Regional aerobiology can help in diagnosis and treatment of allergic rhinitis. Environmental factors contribute to a high concentration of aeroallergen that led to the increased allergy cases among the residents of Texas Panhandle. Allergy and Asthma cases have doubled in the Texas Panhandle area since 2007 (Ranaivo, 2011). Aeroallergens cause serious allergic and asthmatic reactions. Analyzing the aeroallergens with a Burkard Spore Trap provided information regarding the onset, duration, and severity of the pollen season that clinicians use to guide allergen selection for skin testing and treatment. We have been investigating the daily aeroallergen concentration in terms of the meteorological conditions such as daily temperature, wind speed and precipitation. We used a Burkard Volumetric Spore Trap to determine the daily aeroallergen index by collecting aeroallergen samples and characterizing them for 15 years. In the Burkard Volumetric Spore Trap the drum rotates while a vacuum pump sucks in the air trapping the aeroallergen onto the tape. Exposed Melinex tape was stained, mounted and observed under a BX-40 Olympus microscope attached to a DP-70 digital camera and a computer with the Image Pro plus software. The aeroallergens were micro-graphed and the monthly aeroallergen data were compared with the incidence of allergy and asthma cases from 2000-2014. Aeroallergens were viewed for fluorescence with FITC and TRITC fluorescent filters, micrographed and analyzed. A high-pressure
mercury lamp was used to excite the storage molecules or proteins, which exhibited autofluorescence. SEM proved to be useful for observing ultrastructural details like pores, colpi, sulci and ornamentations on the pollen surface. Pollen grains were measured under SEM using the TM-1000 imaging software that revealed the size of colpi or sulci and the distance between the micro-structures. The aeroallergen data that we collected using a Burkard Spore Trap for 15 years showed a steady increase in aeroallergen concentration in the Texas Panhandle area. A fluctuation and gradual shift in aeroallergen index with the warmer climate and a shift in flowering seasons were noticed that contributed to the increased allergy cases. The characterization and analysis of microscopic aeroallergens was accomplished using Fluorescent and Scanning Electron Microscopy. Aeroallergens that were viewed, recorded, and analyzed with fluorescent microscopy exhibited storage protein, oil granules, and the layer of sporopollenin, along with additional ultra-structural details like concordant pattern, exines, pores, colpi, sulci, and other ornamentations. The SEM provided micro-measurements and additional views of the detailed ultrastructural morphology. Analyzing the aeroallergens collected and sampled with the Burkard Spore Trap provided information regarding the onset, duration, and severity of the pollen season that was compared to the number of patient cases seen over a 15-year period. The data accumulated for these studies can be utilized for the forecasting the types and duration of the pollen season. Temperature was found to have an inverse relationship with mold spore concentration. Rainfall had a direct correlation with the mold count directly, increase in precipitation resulted in subsequent higher mold spore concentrations.
