Effects of the addition of endotoxin during perfusion of isolated forelimbs of equine cadavers

Bianca Patan-Zugaj Department for Horses and Small Animals, Veterinary University Vienna, A-1210 Vienna, Austria.

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Felicia C. Gauff Department for Horses and Small Animals, Veterinary University Vienna, A-1210 Vienna, Austria.

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Theresia F. Licka Department for Horses and Small Animals, Veterinary University Vienna, A-1210 Vienna, Austria.
Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, EH25 9RG, Scotland.

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Abstract

Objective—To examine the effect of endotoxins on metabolism and histopathologic changes of isolated perfused equine forelimbs.

Sample—Forelimbs (comprising the metacarpus and digit) were collected from cadavers of 12 healthy adult horses after slaughter at an abattoir (14 limbs; 1 forelimb of 10 horses and both forelimbs of 2 horses).

Procedures—Forelimbs were perfused for 10 hours with autologous blood, with and without the addition of endotoxin (80 ng of lipopolysaccharide [LPS]/L). Two limbs of the endotoxin exposure group and 2 nonperfused limbs were loaded to failure of the suspensory apparatus of the pedal bone to evaluate the effect of body weight. Metabolic and histologic variables were evaluated.

Results—Blood pressure increased during the first hour and did not differ between groups. Lactate dehydrogenase activity was similar in both groups and increased significantly during the 10-hour period; glucose consumption at 5 hours and lactate concentration at 8 hours were significantly higher in limbs exposed to endotoxin. The width of secondary epidermal lamellae was greater in LPS limbs. In the primary dermal lamellae of LPS limbs, there were significantly more vessels with an open lumen and aggregates of intravascular neutrophils.

Conclusions and Clinical Relevance—In the blood-perfused isolated forelimbs of equine cadavers, exposure to LPS led to significant changes in the laminar tissue as well as to metabolic changes. Therefore, endotoxin should be considered as a causative factor for laminitis and not merely as a risk factor.

Abstract

Objective—To examine the effect of endotoxins on metabolism and histopathologic changes of isolated perfused equine forelimbs.

Sample—Forelimbs (comprising the metacarpus and digit) were collected from cadavers of 12 healthy adult horses after slaughter at an abattoir (14 limbs; 1 forelimb of 10 horses and both forelimbs of 2 horses).

Procedures—Forelimbs were perfused for 10 hours with autologous blood, with and without the addition of endotoxin (80 ng of lipopolysaccharide [LPS]/L). Two limbs of the endotoxin exposure group and 2 nonperfused limbs were loaded to failure of the suspensory apparatus of the pedal bone to evaluate the effect of body weight. Metabolic and histologic variables were evaluated.

Results—Blood pressure increased during the first hour and did not differ between groups. Lactate dehydrogenase activity was similar in both groups and increased significantly during the 10-hour period; glucose consumption at 5 hours and lactate concentration at 8 hours were significantly higher in limbs exposed to endotoxin. The width of secondary epidermal lamellae was greater in LPS limbs. In the primary dermal lamellae of LPS limbs, there were significantly more vessels with an open lumen and aggregates of intravascular neutrophils.

Conclusions and Clinical Relevance—In the blood-perfused isolated forelimbs of equine cadavers, exposure to LPS led to significant changes in the laminar tissue as well as to metabolic changes. Therefore, endotoxin should be considered as a causative factor for laminitis and not merely as a risk factor.

Contributor Notes

Supported in part by a research grant from the Veterinary University Vienna and the Austrian Science Fund (FWF Project No. P22598).

The authors thank Magda Helmreich for technical assistance.

Address correspondence to Dr. Licka (theresia.licka@vetmeduni.ac.at).
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