Laboratory and clinical evaluation of a chromogenic endotoxin assay for horses with acute intestinal disorders.
Abstract: In this study the laboratory and clinical performance of a chromogenic endotoxin assay for equine plasma was evaluated. The assay was sensitive (detection limit 3 ng LPS/L plasma), reproducible (within and between-assay CV at 50 ng LPS/L E. coli O111:B4 LPS standard addition was 5% and 7.5%, respectively), and not substantially affected by enhancement or inhibition phenomena (recovery of an in vitro spike was 75-125% in 80% of the samples). LPS added to whole blood was rapidly inactivated upon incubation at 37 degrees C but not at 0 degrees C. A recently developed blood collection tube for LPS testing was found suitable, i.e. LPS-free and providing non-contaminated samples. In 48 horses suffering from acute abdominal diseases requiring surgical treatment, LPS levels were significantly higher in platelet-rich plasma (PRP) than in platelet-poor plasma (PPP), and the proportional difference was related to the PRP platelet count (r = 0.52, p < 0.001, mean difference 48%, range 8-77%). LPS levels were also significantly higher in horses that died or were euthanized than in surviving horses (mean 16.5 and 7.1 ng/L PRP, respectively, p < 0.05). We conclude that LPS can be measured in equine plasma with picogram sensitivity and recommend the use of PRP instead of PPP for clinical LPS testing. For clinical use a decision limit for endotoxaemia of 5 ng LPS/L PRP appeared to be inadequate. Analysis at a higher cut-off level for endotoxaemia and the evaluation of clinical, pathological, and laboratory parameters would be more meaningful.
Publication Date: 1994-05-01 PubMed ID: 7801503
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- Journal Article
Summary
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The research investigates the effectiveness of a chromogenic endotoxin assay (test) used for detecting levels of endotoxin, a component of some harmful bacteria, in the blood of horses suffering from acute abdominal disorders. It concludes that the test shows high sensitivity and recommends it for clinical use, but further suggests modifications to the interpretation of its results.
Overview of Research Methodology
- The sensitivity and reproducibility of a chromogenic endotoxin assay were tested. The goal was to determine whether this assay could effectively measure endotoxin levels in horse plasma (a component of blood).
- The assay’s sensitivity – its ability to detect low levels of endotoxins – was determined to be 3 ng LPS/L plasma.
- It was also found to be reproducible, meaning that the test could return similar results under consistent conditions. Specifically, it had a 5% within-assay coefficient of variation (CV) and a 7.5% between-assay CV for a 50 ng/L endotoxin (E. coli LPS) standard addition.
- The research also observed whether the assay was significantly affected by enhancement or inhibition phenomena and found it to have 75-125% recovery in 80% of the samples.
Evaluation of Blood Collection Tube and Different Plasmas
- The paper examined the utility of blood collection tubes for LPS testing, verifying them to be suitable as they were LPS-free and provided non-contaminated samples.
- In clinical examinations, LPS levels in the blood of 48 horses suffering from acute abdominal diseases were found to be significantly higher in platelet-rich plasma (PRP) than in platelet-poor plasma (PPP).
- The difference in LPS levels between the two types of plasma was related to the PRP platelet count.
Outcomes in Relation to Survival Rates
- The study found that LPS levels were significantly higher in horses that died or were euthanized, compared to surviving horses.
- This information indicates that high levels of endotoxins in plasma may contribute to worse outcomes in horses with acute abdominal diseases.
Conclusions and Recommendations
- The study concludes that this chromogenic endotoxin assay can effectively be used to measure LPS in equine plasma.
- While PRP is recommended for clinical LPS testing, the researchers suggest that the current decision limit of 5 ng LPS/L PRP may not be sufficient. Higher cut-off levels for endotoxemia were recommended. To make the results more meaningful, researchers advised evaluating additional clinical, pathological, and laboratory parameters.
Cite This Article
APA
Steverink PJ, Salden HJ, Sturk A, Klein WR, van der Velden MA, Németh F.
(1994).
Laboratory and clinical evaluation of a chromogenic endotoxin assay for horses with acute intestinal disorders.
Vet Q, 16 Suppl 2, S117-S121.
Publication
Researcher Affiliations
- Department of General and Large Animal Surgery, Faculty of Veterinary Medicine, Utrecht University, The Netherlands.
MeSH Terms
- Acute Disease
- Animals
- Chromogenic Compounds
- Horse Diseases / blood
- Horses
- Intestinal Diseases / blood
- Intestinal Diseases / veterinary
- Lipopolysaccharides / blood
- Sensitivity and Specificity
- Time Factors
Citations
This article has been cited 7 times.- Abdelfatah SH, Yassin AM, Khattab MS, Abdel-Razek AS, Saad AH. Spirulina platensis as a growth booster for broiler; Insights into their nutritional, molecular, immunohistopathological, and microbiota modulating effects. BMC Vet Res 2024 Jan 5;20(1):11.
- Ehrmann C, Engel J, Moritz A, Roscher K. Assessment of platelet biology in equine patients with systemic inflammatory response syndrome. J Vet Diagn Invest 2021 Mar;33(2):300-307.
- Taylor S. A review of equine sepsis. Equine Vet Educ 2015 Feb;27(2):99-109.
- MacDonald ES, Barrett JG. The Potential of Mesenchymal Stem Cells to Treat Systemic Inflammation in Horses. Front Vet Sci 2019;6:507.
- Patan-Zugaj B, Egerbacher M, Licka TF. Endotoxin-induced changes in expression of cyclooxygenase isoforms in the lamellar tissue of extracorporeally haemoperfused equine limbs. Anat Histol Embryol 2020 Sep;49(5):597-605.
- Chen J, Tellez G, Richards JD, Escobar J. Identification of Potential Biomarkers for Gut Barrier Failure in Broiler Chickens. Front Vet Sci 2015;2:14.
- Krueger CR, Ruple-Czerniak A, Hackett ES. Evaluation of plasma muscle enzyme activity as an indicator of lesion characteristics and prognosis in horses undergoing celiotomy for acute gastrointestinal pain. BMC Vet Res 2014;10 Suppl 1(Suppl 1):S7.
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