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Home / Equine Research Bank / Appl Environ Microbiol / Evaluation of the Limulus amebocyte lysate and recombinant f...
Applied and environmental microbiology2010; 76(15); 4988-4995; doi: 10.1128/AEM.00527-10

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.
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Publication Date: 2010-06-04 PubMed ID: 20525858PubMed Central: PMC2916455DOI: 10.1128/AEM.00527-10Google Scholar: Lookup
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  • Comparative Study
  • Evaluation Study
  • Journal Article
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  • U.S. Gov't
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study compared the efficacy of two methods, the Limulus amebocyte lysate (LAL) assay and the recombinant factor C (rFC) assay, in measuring airborne endotoxin concentrations. The researchers concluded that both assays provided similar results when assessing endotoxin exposure in livestock facilities.

Understanding Endotoxins, LAL and rFC

  • Endotoxin is a potent inflammatory substance that stimulates immune response. It’s often present in organic dust in livestock facilities, posing potential health risks, such as lung ailments, to those in residential and occupational settings.
  • The Limulus Amebocyte Lysate (LAL) and the Recombinant Factor C (rFC) are methods used to detect and measure endotoxin concentrations.
  • The LAL assay is a traditional method of endotoxin detection, based on the blood cells (amebocytes) of the horseshoe crab, Limulus polyphemus.
  • Recombinant Factor C (rFC) is a synthetically produced equivalent of a factor in the Limulus pathway. It is used in a fluorometric assay – an advanced approach in endotoxin detection.

Research Process and Findings

  • The researchers collected parallel air samples in both the field (at livestock production facilities) and laboratory (using resuspended airborne dust from livestock facilities).
  • They employed different types of samplers for air sampling, including Institute of Occupational Medicine (IOM) samplers, Button samplers, closed-face cassettes, and cyclonic samplers.
  • After the extraction of samples, they analyzed them using both LAL and rFC assays.
  • Across 402 field sample pairs, there was a high degree of correlation between endotoxin concentrations recorded by the LAL and rFC methods (r = 0.93).
  • In 510 laboratory sample pairs, an almost equally strong correlation was observed (r = 0.86).
  • The correlations varied depending on the facility or dust type, ranging from 0.65 to 0.96.
  • An analysis of variance (ANOVA) revealed significant interactions of facility-sampler and facility-assay.
  • The LAL and rFC assays returned closely aligned results, reaffirming the utility of both methods in assessing endotoxin exposure in livestock facilities.

Implications and Conclusions

  • This study backed up both the Limulus amebocyte lysate (LAL) and recombinant factor C (rFC) assays as reliable methods for assessing airborne endotoxin.
  • The two assays indicated similar exposure estimates, demonstrating comparable performance.
  • The results highlight the possibility of incorporating both methods into future studies or assessments for airborne endotoxin concentrations in various environments.

Cite This Article

APA
Thorne PS, Perry SS, Saito R, O'Shaughnessy PT, Mehaffy J, Metwali N, Keefe T, Donham KJ, Reynolds SJ. (2010). Evaluation of the Limulus amebocyte lysate and recombinant factor C assays for assessment of airborne endotoxin. Appl Environ Microbiol, 76(15), 4988-4995. https://doi.org/10.1128/AEM.00527-10

Publication

Applied and environmental microbiology
ISSN: 1098-5336
NlmUniqueID: 7605801
Country: United States
Language: English
Volume: 76
Issue: 15
Pages: 4988-4995

Researcher Affiliations

Thorne, Peter S
  • Department of Occupational and Environmental Health, University of Iowa, Iowa City, Iowa 52242, USA. peter-thorne@uiowa.edu
Perry, Sarah S
    Saito, Rena
      O'Shaughnessy, Patrick T
        Mehaffy, John
          Metwali, Nervana
            Keefe, Thomas
              Donham, Kelley J
                Reynolds, Stephen J

                  MeSH Terms

                  • Air / analysis
                  • Air Microbiology
                  • Animals
                  • Animals, Domestic
                  • Arthropod Proteins
                  • Bacteriological Techniques / methods
                  • Endotoxins / analysis
                  • Enzyme Precursors / metabolism
                  • Housing, Animal
                  • Limulus Test / methods
                  • Sensitivity and Specificity
                  • Serine Endopeptidases / metabolism

                  Grant Funding

                  • 5U50OH07548 / NIOSH CDC HHS
                  • P30 ES005605 / NIEHS NIH HHS
                  • U50 OH008085 / NIOSH CDC HHS
                  • R01 OH007841 / NIOSH CDC HHS
                  • U50 OH007548 / NIOSH CDC HHS
                  • P30ES05605 / NIEHS NIH HHS
                  • 5U50OH008085 / NIOSH CDC HHS
                  • R01OH007841 / NIOSH CDC HHS

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                  Citations

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