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Evaluation of the Limulus amebocyte lysate and recombinant factor C assays for assessment of airborne endotoxin.

ABSTRACT: As a potent inflammatory agent, endotoxin is a key analyte of interest for studies of lung ailments in domestic environments and occupational settings with organic dust. A relatively unexplored advance in endotoxin exposure assessment is the use of recombinant factor C (rFC) from the Limulus pathway in a fluorometric assay. In this study, we compared airborne endotoxin concentrations in laboratory- and field-collected parallel air samples using the kinetic Limulus amebocyte lysate (LAL) assay and the rFC assay. Air sampling was performed using paired Institute of Occupational Medicine (IOM) samplers, Button samplers, closed-face cassettes, and cyclone samplers. Field sampling was performed in 10 livestock production facilities, including those housing swine, chicken, turkey, dairy cows, cattle, and horses. Laboratory sampling was performed in exposure chambers using resuspended airborne dust collected in five livestock facilities. Paired samples were extracted in pyrogen-free water with 0.05% Tween 20 and analyzed using LAL and rFC assays. In 402 field sample pairs there was excellent agreement between endotoxin concentrations determined by LAL and rFC (r = 0.93; P < 0.0001). In 510 laboratory sample pairs there was also excellent agreement between the two assays (r = 0.86; P < 0.0001). Correlations for subgroups of facility or dust type ranged from 0.65 to 0.96. Mixed-model analysis of variance (ANOVA) for the field studies showed significant interactions of facility-sampler and facility-assay. rFC/LAL ratios of the geometric means were 0.9 to 1.14 for the samplers (not significantly different from 1.0). The data from this study demonstrate that the LAL assay and the rFC assay return similar estimates of exposure in livestock facilities. Both methods provided suitable lower limits of detection such that all but 19 of 1,824 samples were quantifiable.

SUBMITTER: Thorne PS

PROVIDER: S-EPMC2916455 | biostudies-literature | 2010 Aug

REPOSITORIES: biostudies-literature

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Evaluation of the Limulus amebocyte lysate and recombinant factor C assays for assessment of airborne endotoxin.

Thorne Peter S PS Perry Sarah S SS Saito Rena R O'Shaughnessy Patrick T PT Mehaffy John J Metwali Nervana N Keefe Thomas T Donham Kelley J KJ Reynolds Stephen J SJ

Applied and environmental microbiology 20100604 15

As a potent inflammatory agent, endotoxin is a key analyte of interest for studies of lung ailments in domestic environments and occupational settings with organic dust. A relatively unexplored advance in endotoxin exposure assessment is the use of recombinant factor C (rFC) from the Limulus pathway in a fluorometric assay. In this study, we compared airborne endotoxin concentrations in laboratory- and field-collected parallel air samples using the kinetic Limulus amebocyte lysate (LAL) assay an ...[more]

PMID: 20525858

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Project description:BACKGROUND:Elevated endotoxin levels have been measured in ambient air around livestock farms, which is a cause of concern for neighbouring residents. There is clear evidence that occupational exposure to high concentrations of airborne endotoxin causes respiratory inflammation, respiratory symptoms and lung function decline. However, health effects of exposure to low levels of endotoxin are less well described. The aim of this systematic review is to summarize published associations between exposure to relatively low levels of airborne endotoxin and respiratory health endpoints. METHODS:Studies investigating respiratory effects of measured or modelled exposure to low levels of airborne endotoxin (average < 100 EU/m3) were eligible for inclusion. In total, 1362 articles were identified through a Pubmed database search, of which 31 articles were included in this review. Studies were included up to February 2017. Overview tables and forest plots were created, and study quality was assessed. RESULTS:Twenty-two included studies had a cross-sectional design, others were designed as longitudinal observational (n = 7) or experimental (n = 2) studies. Most studies (n = 23) were conducted in an occupational setting, some involved domestic or experimental exposure. Several studies reported statistically significant effects of exposure to low levels of endotoxin on respiratory symptoms and lung function. However, considerable heterogeneity existed in the outcomes of the included studies and no overall estimate could be provided by meta-analysis to quantify the possible relationship. Instead, a best evidence synthesis was performed among studies examining the exposure-response relationship between endotoxin and respiratory outcomes. Significant exposure-response relationships between endotoxin and symptoms and FEV1 were shown in several studies, with no conflicting findings in the studies included in the best evidence synthesis. Significantly different effects of endotoxin exposure were also seen in vulnerable subgroups (atopics and patients with broncho-obstructive disease) and smokers. CONCLUSIONS:Respiratory health effects of exposure to low levels of airborne endotoxin (< 100 EU/m3) seem plausible. Future studies are needed to investigate ambient exposure to endotoxin and potential respiratory health effects, especially in vulnerable subgroups of the population.

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Evaluation of the Limulus amebocyte lysate and recombinant factor C assays for assessment of airborne endotoxin.

Publications

Evaluation of the Limulus amebocyte lysate and recombinant factor C assays for assessment of airborne endotoxin.

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