Comparison of endotoxin assays using agricultural dusts

Endotoxins from gram-negative bacteria pose a significant respiratory hazard. Establishing dose-response relationships is problematic because there are no standard procedures for sampling and analysis. The goal of this study was to compare endotoxin analyses in six laboratories using Limulus-based assays for analysis of organic dusts from three agricultural environments: chicken barns, swine barns, and corn processing facilities. For each dust generation experiment 14 side-by-side air samples were collected on 37-mm glass fiber filters at flows of 1.8 L/min. Each laboratory was randomly allocated two filters from each of seven experiments per dust type. Three laboratories used the QCL-1000 endpoint assay, and three used the kinetic-QCL method. To eliminate variability among different lots, a single lot of Limulus amebocyte lysate for endpoint assays and one similar lot for kinetic assays was provided. Precision of assays performed within labs was very good, with pooled coefficients of variation for replicate samples ranging from 1 to 11% over all labs and all dust types. There were significant differences between laboratories for all three dust types (p < 0.01). The pattern of differences between labs varied by dust type. For chicken dust, labs using the endpoint method reported higher results than those using kinetic methods. For swine and corn dusts, labs using the kinetic method reported the highest endotoxin values. For chicken dust, results from all labs except A and B were highly correlated (r = 0.86-1.00). For swine dust, only labs B and E, and C and D were correlated. For corn, A, B, and D were significantly correlated with most other labs. In conclusion, statistical differences in performance between laboratories were apparent and may be related to the extraction and analytical methods. The results of this study will be useful for standardization of sampling and analysis of airborne endotoxin in agriculture.