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The Limulus Amoebocyte Lysate Test: Detection of Endotoxin in Plasma of Swine and Cattle

Limulus Amøbocyt Lysat testen: Påvisning af endotoksin i svine- og kvægplasma. IL In vivo undersøgelser

II. In Vivo Investigations

Acta Veterinaria Scandinavica volume 26, pages 246–261 (1985)Cite this article

Abstract

Endotoxin was detected by the LAL test in the plasma of swine and cattle following in vivo injection of endotoxin in order to evaluate the applicability of the test in veterinary medicine for detection of endotoxemia.

Clinical symptoms of endotoxemia occurred after the injection into 3 swine of 0.10–0.25 mg endotoxin/100 kg bwt and after the infusion during 1–1½ h of 2.0–2.2 mg endotoxin/100 kg bwt into 3 calves. The concentration of endotoxin detected by the LAL test in the experimental animals ranged from 0.15 ng to 6.0 ng endotoxin per ml crude plasma. As positive LAL reactions were obtained only in close connection to the administration of endotoxin, clearance of endotoxin to levels below the sensitivity of the test was fast. In spite of the fast clearance, light symptoms of endotoxemia could be seen as long as 24 h after the last sample showing a positive test result. The applied technique for LAL analysis on blood, therefore, was not adequate for detection of endotoxin at sufficiently low concentrations and some possibilities of improving the technique are discussed.

Though leukocytosis were found not to influence the outcome of the LAL test on blood, leukocytic mediators released by endotoxin or endotoxin-derived injuries may still have caused the persistence of the symptoms of endotoxemia, and this question is disputed in relation to the benefit of improving the technique.

Especielly in the veterinary clinic, great precautions are necessary to obviate false positive test samples resulting from, e.g., external contamination or transient stress caused by excitement, and it is concluded that the possible application of the LAL test for detection of endotoxemia in veterinary medicine is restricted to surveillance of hospitalized animals and to research purposes.

Sammendrag

Ved hjælp af LAL testen blev endotoksin påvist i svine- og kvægplasma efter in vivo injektion med henblik på at vurdere anvendeligheden af testen i veterinærmedicinen til påvisning af endotoksinæmi.

Kliniske symptomer på endotoksinæmi opstod efter injektion af 0,10–0,25 mg endotoxin/100 kg lgv. i 3 svin og efter 1–1½ times infusion af 2,0–2,2 mg endotoksin/100 kg lgv. i 3 kalve. Koncentrationen of endotoksin, der blev påvist ved hjælp af LAL testen i forsøgsdyrene, varierede fra 0,15 ng til 6,0 ng endotoksin pr. ml ubehandlet plasma. På grund af en hurtig clearance blev koncentration af endotoksin hurtigt lavere end testens følsomhed, hvorfor positive LAL reaktioner kun forekom i nær tilknytning til administrationen af endotoksin. På trods af den hurtige clearance kunne lette symptomer på endotoksinæmi ses så længe som 24 timer efter den sidste positive prøve. Derfor var den anvendte teknik til LAL analyse af blod ikke god nok til påvisning af endotoksin i tilstrækkelig lave koncentrationer, og nogle muligheder for at forbedre teknikken diskuteres.

Selv om det blev fundet, at leukocytose ikke influerede på udfaldet af LAL testen på blod, kan de vedvarende symptomer på endotoksinæmi stadig være forårsaget enten af mediatorer frigjort fra leukocyter under endotoksinæmien eller af endotoksin-betingede skader, og dette spørgsmål drøftes i relation til den fordel, der kan opnås ved at forbedre teknikken.

Især i den veterinære klinik er det nødvendigt med mange foranstaltninger for at undgå falske positive prøver, der f. eks. kan skyldes kontamination med endotoksin eller forbigående stress i forbindelse med ophidselse, og det konkluderes, at den mulige anvendelse af LAL testen til påvisning af endotoksinæmi i veterinærmedicinen er begrænset til overvågning af hospitalsindlagte dyr og til forskningsrelaterede formål.

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  1. Institute for Animal Reproduction and the Institute of Surgery, Royal Veterinary and Agricultural University, Bülowsvej 13, DK-1870, Copenhagen V, Denmark

    M. Binder & K. Mortensen

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Binder, M., Mortensen, K. The Limulus Amoebocyte Lysate Test: Detection of Endotoxin in Plasma of Swine and Cattle. Acta Vet Scand 26, 246–261 (1985). https://doi.org/10.1186/BF03546555

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Acta Veterinaria Scandinavica

ISSN: 1751-0147