Surfactant alterations in horses with recurrent airway obstruction at various clinical stages

Undine Christmann Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech and University of Maryland, Blacksburg, VA 24061.

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R. Duncan Hite Section of Pulmonary, Critical Care, Allergy and Immunologic Diseases Medicine, Department of Internal Medicine, School of Medicine, Wake Forest University, Winston-Salem, NC 27157.

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Rachel H. H. Tan Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech and University of Maryland, Blacksburg, VA 24061.

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Craig D. Thatcher Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech and University of Maryland, Blacksburg, VA 24061.

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Sharon G. Witonsky Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech and University of Maryland, Blacksburg, VA 24061.

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Stephen R. Werre Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech and University of Maryland, Blacksburg, VA 24061.

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Virginia A. Buechner-Maxwell Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech and University of Maryland, Blacksburg, VA 24061.

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Abstract

Objective—To evaluate the phospholipid composition and function of surfactant in horses with recurrent airway obstruction (RAO) at various clinical stages and compare these properties with findings in horses without RAO.

Animals—7 horses with confirmed RAO and 7 without RAO (non-RAO horses).

Procedures—Pairs of RAO-affected and non-RAO horses were evaluated before, during, and after exposure to hay. Evaluations included clinical scoring, lung function testing, airway endoscopy, and bronchoalveolar lavage fluid (BALF) absolute and differential cell counts. Cell-free BALF was separated into crude surfactant pellet and supernatant by ultracentrifugation, and phospholipid and protein concentrations were determined. Phospholipid composition of crude surfactant pellets and surface tension were evaluated with high-performance liquid chromatography and a pulsating bubble surfactometer, respectively. Findings were compared statistically via mixed-effects, repeated-measures ANOVA.

Results—Total phospholipid concentration in BALF was lower in RAO-affected versus non-RAO horses at all sample collection times. In the RAO-affected group, total phospholipid concentration was lower during exposure to hay than before or after exposure. There were no significant differences in BALF protein concentration, percentages of phospholipid classes, or surface tension between or within groups of horses.

Conclusions and Clinical Relevance—All clinical stages of RAO-affected horses were characterized by low surfactant concentration in BALF. Exacerbation of RAO led to an additional decrease in surfactant concentration. Causes for low surfactant concentration in RAO-affected horses remain to be determined. Low phospholipid concentration may render RAO-affected horses more susceptible than unaffected horses to surfactant alterations and contribute to clinical disease status and progression.

Abstract

Objective—To evaluate the phospholipid composition and function of surfactant in horses with recurrent airway obstruction (RAO) at various clinical stages and compare these properties with findings in horses without RAO.

Animals—7 horses with confirmed RAO and 7 without RAO (non-RAO horses).

Procedures—Pairs of RAO-affected and non-RAO horses were evaluated before, during, and after exposure to hay. Evaluations included clinical scoring, lung function testing, airway endoscopy, and bronchoalveolar lavage fluid (BALF) absolute and differential cell counts. Cell-free BALF was separated into crude surfactant pellet and supernatant by ultracentrifugation, and phospholipid and protein concentrations were determined. Phospholipid composition of crude surfactant pellets and surface tension were evaluated with high-performance liquid chromatography and a pulsating bubble surfactometer, respectively. Findings were compared statistically via mixed-effects, repeated-measures ANOVA.

Results—Total phospholipid concentration in BALF was lower in RAO-affected versus non-RAO horses at all sample collection times. In the RAO-affected group, total phospholipid concentration was lower during exposure to hay than before or after exposure. There were no significant differences in BALF protein concentration, percentages of phospholipid classes, or surface tension between or within groups of horses.

Conclusions and Clinical Relevance—All clinical stages of RAO-affected horses were characterized by low surfactant concentration in BALF. Exacerbation of RAO led to an additional decrease in surfactant concentration. Causes for low surfactant concentration in RAO-affected horses remain to be determined. Low phospholipid concentration may render RAO-affected horses more susceptible than unaffected horses to surfactant alterations and contribute to clinical disease status and progression.

Contributor Notes

Dr. Tan's present address is School of Veterinary and Biomedical Sciences, James Cook University, Townsville, QLD 4811, Australia.

Dr. Thatcher's present address is School of Applied Arts and Sciences, Arizona State University, Mesa, AZ 85212.

Dr. Chistmann's present address is CIRALE – Equine Sports Medicine, RD 675, 14430 Goustranville, France.

Supported by the American College of Veterinary Internal Medicine Foundation and Morris Animal Health Foundation (fellowship grant).

Presented in part at the 24th Meeting of the Veterinary Comparative Respiratory Society, Jena, Germany, October 2006; the 25th Annual American College of Veterinary Internal Medicine Forum, Seattle, June 2007; and the 3rd European College of Equine Internal Medicine Conference, Barcelona, Spain, January 2009.

The authors thank Barbara Dryman and Bonnie Grier for assistance with sample analysis.

Address correspondence to Dr. Christmann (chrisun@vt.edu).
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