We describe a new microtiter immunospore trapping device (MTIST device) that uses a suction system to directly trap air particulates by impaction in microtiter wells. This device can be used for rapid detection and immunoquantification of ascospores of Mycosphaerella brassicicola and conidia of Botrytis cinerea by an enzymelinked immunosorbent assay (ELISA) under controlled environmental conditions. For ascospores of M. brassicicola correlation coefficients (r 2 ) of 0.943 and 0.9514 were observed for the number of MTIST device-impacted ascospores per microtiter well and the absorbance values determined by ELISA, respectively. These values were not affected when a mixed fungal spore population was used. There was a relationship between the number of MTIST device-trapped ascospores of M. brassicicola per liter of air sampled and the amount of disease expressed on exposed plants of Brassica oleracea (Brussels sprouts). Similarly, when the MTIST device was used to trap conidia of B. cinerea, a correlation coefficient of 0.8797 was obtained for the absorbance values generated by the ELISA and the observed number of conidia per microtiter well. The relative collection efficiency of the MTIST device in controlled plant growth chambers with limited airflow was 1.7 times greater than the relative collection efficiency of a Burkard 7-day volumetric spore trap for collection of M. brassicicola ascospores. The MTIST device can be used to rapidly differentiate, determine, and accurately quantify target organisms in a microflora. The MTIST device is a portable, robust, inexpensive system that can be used to perform multiple tests in a single sampling period, and it should be useful for monitoring airborne particulates and microorganisms in a range of environments.
Detection and enumeration of air-borne spores of fungal plant pathogens has generally been achieved by microscopic examination of surfaces on which spores have been impacted and conventional agar-plating techniques. However, this requires considerable amounts of time and expertise. Such methods are inaccurate for obligate pathogens and unrealistic where there is no selective medium or where there are spore types of similar morphology (such as those produced by ascosporic fungi). In field disease-transmission studies, the use of a Burkard 7-day volumetric spore trap combined with an immunofluorescence test has, however, enabled the detection and quantification of field-trapped ascosporic inoculum of Mycosphaerellu brassicicola (the ringspot pathogen of brassicas). This test has also been found useful in the validation of more rapid user-friendly immunoassay-based trapping procedures. A microtitre immunospore trapping device, which uses a suction system to trap air particulates directly by impaction into microtitre wells, has been used successfully for the rapid detection and quantification of ascosporic inoculum of M. brassicicola. The system shows potential for field detection of air-borne ascosporic inoculum of the ringspot pathogen.
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