J Vet Intern Med 1998;12:163-170

Pulmonary Eosinophilia Associated with Increased Airway

Responsiveness in Young Racing Horses
Jonathan E. Hare and Laurent Viel

Horses are known to acquire small airway disease (SAD), an allergen-induced naturally occurring syndrome of reversible obstruc-
tive lung disease accompanied by airway hyperresponsiveness and increased inflammatory cell numbers on bronchoalveolar lavage
(BAL). This disorder has received scant attention in young racehorses. The purpose of the present report was to examine the effect
of BAL eosinophilia in young racehorses on clinical examination, BAL, hematology, airway responsiveness, and on pulmonary
function at rest and after a standardized exercise challenge. Five (3 males, 2 females; age 2.6 -t 0.9 years) with a history of
respiratory compromise and BAL eosinophil differential count > 5% and 6 controls (4 males, 2 females; age 3.5 2 1.0 years)
training and performing to expectation with normal BAL cell differential (eosinophils < 1%) were studied. Respiratory system
clinical examination was performed and expressed as a clinical score. Arterial blood gas measurements, CBC, and pulmonary
function testing were performed at rest. Pulmonary mechanics measurements were repeated 1 hour and 20 hours after a standardized
treadmill exercise challenge. Incremental histamine inhalation challenge was performed and the concentration of histamine effecting
a 35% decrease in dynamic compliance (PC35Cl,,,) was determined. Significant differences were noted between and controls with
regard to clinical score ( P = .Ol), blood eosinophils ( P = .04), BAL cell count ( P = .04), BAL macrophage differential ( P =
.04), PC35Cl,,, ( P = .008), and tidal volume and respiratory rate at 20 hours following exercise challenge ( P = .05). We conclude
that pulmonary eosinophilia and airway hyperresponsiveness are manifest in some young horses without overt airway obstruction
at rest. We speculate that these may be early events in the natural progression of heaves.
Key words: Lung.

S mall airway disease (SAD) is a commonly recognized

respiratory disability in the horse. Affected individuals
demonstrate recurrent periods of airway obstruction asso-
ciated with increased nonspecific airway responsiveness. I
This syndrome is characterized clinically by coughing, ex-
piratory dyspnea, and an inability to perform maximally
due to respiratory embarrassment.’ Affected individuals
also exhibit varying degrees of nasal discharge, wheezes,
and expanded lung fields on physical examination. The dis-
ease is thought to be an allergic condition because chroni-
cally affected or “heavey” horses demonstrate clinical
signs of obstructive lung disease after exposure to environ-
mental allergens, most commonly hay and straw molds and
fungi.* During exacerbation of their condition, affected
horses demonstrate characteristic reductions in arterial ox-
ygen pressure (Pao,) and pulmonary dynamic compliance
(C), as well as increases in transpulmonary pressure
(APPl),lung resistance (R,), and nonspecific airway hyper-
responsiveness.’
In the past, chronic SAD was considered to be a condi-
tion of the older horse? Recent work, however, has sug-
gested that mild to subclinical SAD is a major cause of
morbidity in the young performance horse and consequent-
ly is a condition of considerable economic concern in the
equine racing industry.5.h
Histologically, the disease is manifest as a bronchiolitis
with varying degrees of smooth muscle hyperplasia, mucus
hypersecretion, epithelial desquamation, and inflammatory
cell infiltration.’ Equine SAD commonly is associated with
~~~
From the Department of Clinicul Studies, Ontario Veterinary Col-
lege, University of Guelph. Guelph, ON, Canada.
Reprint requests: Laurent Viel, Phd, Department of Clinical Studies,
Ontario Veterinary College, University of Guelph, Guelph, ON NIG
2WI, Canada; e-mail: lviel@uoguelph.ca.
Accepted September 30, 1997.
Copyright 0 1998 by the American College of Veterinaty Internal
Medicine
0891-6640/98/1203-0006/$3.00/0
an influx of neutrophils on bronchoalveolar lavage (BAL)
c y t ~ l o g y , although
~.~ other inflammatory cells, particularly
mast cellsh and eosinophils,’.“’ have been implicated in the
pathophysiology of the condition, Observations in our lab-
oratory have identified a subset of young racing horses with
a predominantly eosinophilic population in BAL fluid, a
situation similar to that in human patients with asthma.l’
Approximately 50% of asthmatic human patients dem-
onstrate exacerbation of their disease in response to modest
exercise.Iz We sought to determine if this phenomenon oc-
curred in horses with SAD, given that exercise appears to
be one of the factors that initiates clinical signs of the dis-
ease.
In summary, the purpose of the present study was to
evaluate the effect of pulmonary disease characterized by
BAL fluid eosinophilia in a group of young racing horses
on clinical examination, BAL, hematology, and airway re-
sponsiveness as well as its effect on pulmonary function at
rest and after a standardized exercise challenge.
Materials and Methods
Experimental Animals
Eleven Standardbred horses in full training or racing condition were
selected for inclusion in the trial. All horses were studied under guide-
lines established by the Canadian Council on Animal Care.
Five horses, designated as principals (3 males, 2 females; age 2.6
2 0.9 years), had clinical signs of exercise intolerance referrable to
the respiratory system in the 2 weeks preceding examination. Clinical
signs of poor exercise tolerance included prolonged respiratory recov-
ery, cough, and mucopurulent nasal discharge after exercise or respi-
ratory embarrassment at exercise. Horses had exhibited at least 2 of
the preceding signs in the 2 weeks before the trial. Horses were iden-
tified from the referral population presented to the Ontario Veterinary
College as well as in the practice territories of equine veterinarians in
southwestern Ontario. A full physical examination was performed on
each of these horses to eliminate nonrespiratory causes of poor per-
formance. Upper airway endoscopy was performed at rest, without
sedation, to eliminate pharyngeal and laryngeal anomalies as the cause
of poor performance. All horses were on a regular anthelmintic pro-

164 Hare and Vie1
Table 1. Clinical scoring of respiratory examination. The maximum possible score is 15.
Expiratory effort 0 = normal
Respiratory rate 0 = 512/minute
Nasal discharge 0 = none to thin serous
Submandibular lymph nodes 0 = normal
Thoracic auscultation on rebreathing 0 = normal breath
sounds
Thoracic percussion 0= normal
Cough 0 = none during exam
Bronchoscopic appearance of airways
Airway secretion 0 = none
Airway edema 0 = none
gram and had been treated with ivermectin (Eqvalan, Merck Agvet,
Kirkland, PQ, Canada) in the 2 months preceding the trial. Six horses,
designated as controls (4 males, 2 females; age 3.5 2 1.0 years), had
been racing and training to expectation and were normal on complete
physical examination.
Bronchoalveolar lavage was performed and principal horses were
further selected as those horses having >5% eosinophils on BAL cell
differential count. Control horses had BAL cell differential counts
within normal ranges as determined previously.6
Signed informed consent was obtained from all owners or agents
associated with the animals in the trial. All horses entering the trial
had adhered to the following drug withdrawal guidelines: bronchodi-
lators, expectorants, sodium cromoglycate, antibiotics, and furosemide:
48 hours; nonsteroidal anti-inflammatory drugs: 72 hours; prednisone:
96 hours; dexamethasone: 7 days; and steroids given as depot prepa-
rations: 45 days.
Experimental Protocol
All horses were acclimatized to hospital surroundings for 24 hours
before study. On the morning of day I , a general and detailed respi-
ratory examination was performed and blood was obtained for arterial
blood gas analysis, CBC and differential, and fibrinogen assay. Feces
also were collected for parasitologic examination. Horses were accli-
matized to walking on the treadmill for 30 minutes on the afternoon
of day I . Bronchoalveolar lavage was performed after introduction to
the treadmill. On day 2, horses were instrumented for pulmonary func-
tion testing (PIT) and baseline pulmonary mechanics measurements
were performed between 1300 and 1400 hours. Treadmill exercise
challenge then was performed. Pulmonary function testing was re-
peated at 30, 60, and 90 minutes and 20 hours post-treadmill exercise.
Histamine inhalation challenge was performed on the morning of day 3.
Respiratory Examination
A physical examination of the respiratory system was performed by
the same clinician on all horses before BAL (Table I). Horses were
assigned a clinical respiratory score based on the method of Hare and
coworkers.6 The maximum possible score that any horse could achieve
was 15.
Hematology
After the initial respiratory examination, venous blood samples were
collected for CBC and differential (in ethylenediaminetetraacetic acid
[EDTA]) and fibrinogen (in sodium citrate) measurement (Monoject
blood collection tubes, Sherwood Medical, St Louis, MO). CBCs were
performed using an automated cell counter (Coulter Counter ZM,
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1 = increased
1 = 12-16/minute 2 = > 16/minute
I = copious serous 2 = mucopurulent
1 = enlarged
1 = crackles or wheezes 2 = crackles or wheezes
on 5 2 sites on >2 sites
1 = increased resonance
in peripheral lung or
reduced resonance in
caudodorsal lung
1 = once during exam 2 = more than once dur-
ing exam
1 = mucoid 2 = mucopurulent
1 = mild blunting of ca- 2 = moderate blunting of
rina carina
Coulter Electronics of Canada Ltd, Burlington, ON, Canada). WBC
differential counts were performed in a routine manner. Plasma fibrin-
ogen concentrations were measured in blood collected in citrate
(FibroSystem, BBL, Cockeysville, MD).
Arterial blood samples were collected anaerobically by direct punc-
ture of the carotid or transverse facial artery using a 20-gauge needle
and heparinized plastic syringe (Bard Parker A-line Kit, Becton Dick-
inson Acute Care, Franklin Lakes, NJ). Blood gas analysis was per-
formed immediately on a Radiometer ABL3 analyzer (Radiometer
Corp, Copenhagen, Denmark).
Parasitology
Feces and BAL fluid were examined for evidence of equine lung-
worm (Dictyocaulus arnjeldi) larvae by the Baermann technique.
Bronchoscopy and BAL
Bronchoscopy and BAL procedures were conducted as previously
de~cribed.~.’ Briefly, the horse was restrained in stocks and a nose
twitch was applied. A 13-mm-diameter 180-cm-long bronchoscope
(Olympus CF-LB2, Olympus Corp, Tokyo, Japan) equipped with a
cold light source (Olympus CLK-3, Olympus Corp) was passed into
the nasal cavity and the nasopharynx was inspected. The bronchoscope
then was withdrawn and the animal was sedated with 0.5 m g k g xy-
lazine (Anased, Lloyd Laboratories, Shenandoah, IA) IV. After seda-
tion the bronchoscope was passed through the naris into the trachea
to the level of the carina and 60 mL of 0.2% lidocaine solution was
sprayed through the biopsy channel to alleviate airway discomfort and
cough. Next, the bronchoscope was wedged in a subsegmental bron-
chus of the right diaphragmatic lung lobe. The lavage consisted of the
infusion and immediate aspiration of 2 250-mL aliquots of warmed
(37°C). sterile physiologic saline. Fluid was retrieved by means of a
medical suction pump (Medi-pump, Power Air Division, Sheboygan,
WI) calibrated to deliver a constant negative pressure of 100 mm Hg.
Aspiration was discontinued when 200 mL of fluid had been retrieved.
Processing of BAL Fluid
The lavage fluid was kept on ice until processing. Concentrations
of nucleated cells in BAL fluid were measured with an automated cell
counter (Coulter Counter ZM). Air-dried slides of BAL fluid were
prepared by cytocentrifuge at 600 rpm for 6 minutes (Cytospin 2,
Shandon Southern Products Ltd, Astmoor, UK). The slide preparations
then were stained with a Wright-Giemsa stain using an automated
stainer (Hema-Tek Slide Stainer, Miles Laboratories Ltd, Rexdale, ON,
Canada). Five hundred consecutive nucleated cells were identified on
each Wright-Giemsa-stained slide for a complete differential count.

Pulmonary Eosinophilia in Young Racing Horses
The investigator was blinded as to the identity of the slides during the
differential cell counts. Calculation of cell concentrations in BAL fluid
was obtained by multiplying the cell differential percentage by the
nucleated cell count. Sixty-milliliter aliquots of BAL fluid were sub-
mitted for parasitologic examination using the Baermann technique.
Pulmonary Function Testing
Horses were restrained with a lead and halter in a 1 X 3-m stock
without sedation. A flexible plastic face mask was fitted over the an-
imal’s muzzle and connected to a heated Fleisch #4 pneumotachograph
(Gould Electronics, Biltholven, The Netherlands), which was used to
measure air flow, with volume derived by the electronic integration of
the flow signal (Pulmonary Mechanics Analyzer, Buxco Electronics
Inc, Sharon, CT). Transpulmonary pressure was measured as the dif-
ference between pressure at the airway opening (facemask) and pres-
sure in a latex esophageal balloon (10 cm long, with 3 mL of air
injected, attached to a 2-m polyethylene catheter, 2.69 mm inner di-
ameter, 3.5 mm outer diameter), inserted to mid-thorax, to estimate
pleural pressure. The frequency responses of the transducer and cath-
eter systems were measured, with amplitude found to be without at-
tenuation up to 20 Hz. The phase lag between the pressure and flow
signals was corrected by using a digital delay system (Buxco Elec-
tronics Inc) The flow and pressure signals were collected and stored
on a personal computer for respiratory loop analysis using software
from Buxco Electronics. Five to 10 breaths from each recording seg-
ment, chosen on the basis of loop closure and uniformity, were aver-
aged and the average parameter values recorded. Flow-volume and
pressure-volume loops were analyzed from the same selected breaths.
Exercise Challenge
A safety harness and heart rate monitor (Equistat model HR-8A)
were attached to the horse and hobbles were fitted prior to exercise
on a treadmill designed for equine use (Sato equine treadmill, Uppsala,
Sweden). Temperature in the treadmill room was maintained at 22°C
and humidity was between 40 and 60%. The horse was led onto the
treadmill and the treadmill was started at its lowest speed (1.5 m/
second) for 3 minutes. The treadmill then was inclined 3” and the
speed increased to 7-8 &second for 2 minutes, then to between 10
and 1 1 d s e c o n d for 3 minutes, the final speed being that at which
the horse maintained a heart rate of approximately 200 beats per min-
ute. After this time, the treadmill was slowly brought to a standstill.
The horse was removed from the treadmill and cooled down by hand-
walking.
Histamine Inhalation Challenge
The method of histamine aerosol provocation was a modification of
that of Mirbahar and coworkers” and Klein and Deegen,I4 and was
based on the work of Cockroft and coworkers.Ir Horses were re-
strained with a lead and halter in a 1 X 3-m stock without sedation.
A flexible plastic face mask was fitted over the animal’s muzzle and
connected to a low resistance nonrebreathing valve to prevent the mix-
ing of inspired and expired gases.Ih Baseline pulmonary function pa-
rameters were recorded prior to aerosolization of a 0.9% saline solu-
tion using a gas flow of 4.5 L/minute generated by a compressor (AFP
Medical, Rugby, UK) through a nebulizer (Mistineb, Wilder Medical,
Kitchener, ON, Canada). At this flow rate, the nebulizer output was
0.13 T 0.05 mL/minute. Each horse was then exposed to aerosoliza-
tion of doubling doses of histamine diphosphate solution (1-32 mg/
mL; Sigma Chemical Co, St Louis, MO). Pulmonary function param-
eters were recorded immediately after each 2-minute aerosolization.
Aerosolization was discontinued when dynamic compliance decreased
to 50% of that recorded after saline aerosolization, a value previously
reported by Derksen and coworkers,l or when transpulmonary pressure
doubled. Data were plottcd against the log of histamine dose and, by
interpolation between the points of the concentration-response curve,
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165
the provocative concentration of histamine that caused a 35% decrease
in dynamic compliance (PC35Cl,,,) was determined. If C,,,,, had de-
creased by less than 35% after 32 mg/mL histamine, PC35Cl,,, was
expressed as 32 mg/mL.
Statistical Analysis
Respiratory score, hematology, BAL, and airway responsiveness
data were compared between principal and control horses by the
Mann-Whitney U-test (Statistix 4.0, Analytical Software, St Paul,
MN).” Correlations between percent BAL fluid eosinophils and other
parameters were determined by Spearman rank correlation (Statistix
4.0).lx The effect of BAL eosinophilia on pulmonary function at base-
line and after exercise challenge was evaluated by repeated measures
analysis of variance using Dunnet’s test for means comparison.“’
Results
The results of the respiratory scores, hematology, BAL
cell constituents, and airway responsiveness data of prin-
cipal and control horses are presented in Table 2. Signifi-
cant differences were observed between principal and con-
trol horses with regard to respiratory score, peripheral blood
eosinophils, BAL total cell count and macrophage percent-
age on differential, and PC35C,,,. Pulmonary function pa-
rameters obtained at different time periods postexercise are
summarized in Table 3. Significant differences were noted
between principal and control horses with regards to both
respiratory rate and tidal volume at 20 hours. The BAL
eosinophil differential count correlated strongly with both
clinical score (Fig 1) and PC35C,,,, (Fig 2).
Discussion
Strong evidence exists to support the hypothesis that air-
way inflammation is associated with increased nonspecific
airway responsiveness in horses’.2oas well as in other spe-
cies.2‘-23In general, it is believed that release of mediators
by the inflammatory cell population serves to increase the
permeability of the respiratory epithelium to noxious
agents, to potentiate the contraction of airway smooth mus-
cle, to increase the sensitivity of neuronal receptors in the
airway, and to increase the thickness of the airway walls
by cellular infiltration and mucus p r o d u ~ t i o nThe . ~ ~ human
eosinophil has received particular attention in this regard
because it is a predominant airway inflammatory cell in
chronic asthma.” Equine eosinophils, like their human
counterparts, are known to contain several cationic proteins
including major basic protein,25.2heosinophil peroxidase,z5
and eosinophil cationic protein.?’ These enzymes are be-
lieved to contribute to a number of airway events including
mast cell degranulation, epithelial damage, and lymphocyte
i n h i b i t i ~ n In
. ~ ~addition, both lipoxygenase and cyclooxy-
genase products of the arachidonic acid pathway are pro-
duced by e o s i n o p h i l ~and , ~ ~ these cells are a major source
of platelet activating factor.2X
In the present study, pulmonary eosinophilia on BAL
cytology was taken as evidence of allergic lung disease.
The presence of airway eosinophilia is not a novel obser-
vation in the h o r ~ e . ~The , ~ condition
~ - ~ ~ has been described
to occur secondary to infection with D. amfeldi, the lung-
worm of horses.2y.30 Other investigators have postulated that
airway eosinophilia may occur secondary to SAD in the
 19391676, 1998, 3, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/j.1939-1676.1998.tb02112.x by Cochrane Colombia, Wiley Online Library on [10/07/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
166 Hare and Vie1

Table 2. Clinical score, hematology, bronchoalveolar lavage, and airway responsiveness results in principal and control
horses. Values are mean 2 SD.
Principals (n = 5) Controls (n = 6) P
(Mann-
Parameter Median Range Median Range Whitney)
Clinical score 4 3-10 I .5 0-3 .o I
Hematology
WBC ( X 10y/L) 10.4 8.4-1 1.O 7.4 5.7-9.5 .06
Eosinophils ( X 109/L) 0.3 0.1-0.9 0.07 04.11 .04
Paoz (mm Hg) 91.0 78.6-96.1 95.8 88.6-102.1 .I7
Bronchoalveolar lavage
Total cells ( X 106iL) 650 320-1,100 360 260-540 .04
Macrophage (%) 58.6 40.6-62.4 67.7 61.0-78.8 .o 1
( X loon) 389 202-501 263 179-339 .06
Lymphocyte (%) 25.8 22.6-3 1.4 31.5 19.0-35.0 .32
( X lOf'/L) 203 74-250 100 52-1 84 .I2
Neutrophil (410) 0.8 0-1.8 0.4 0.2-1.4 .58
( X IOVL) 5.8 0-7.8 1.1 0.8-7.6 .3 I
Eosinophil (%) 11.8 6.4-26.4 0.3 0-1 .o
( X IOVL) 65 38-292 0.8 0-5.4
Mast cell (%) 1.4 0.4-2.4 1.o 0-2.8 36
( X IOVL) 12 3-16 2.8 0-15.2 .I7
Epithelial (%) 0 0-3.2 0 04.6 .93
( X 10") 0 0-2 1 0 0-3.1 .93
Airway responsiveness: PC35C,,,. (m.dmL) 4.4 1-10.7 21.4 1 1.2-32 .o 1
Pao,, arterial oxygen tension: PC35C,,,,, concentration of histamine effecting a 35% decrease in dynamic compliance
Entry variable.

absence of parasitic i n f i l t r a t i ~ n . ~The

, ~ ~ )horses
, ~ ' in the pres-
ent study were found to be negative for parasites on Baer-
mann examination of feces and BAL fluid, suggesting that
the presence of a patent infection with D. arnjeldi was
unlikely. In addition, all animals had been treated recently
with ivermectin, a drug demonstrated to have excellent ef-
ficacy against all life stages of D. ~ r n j e l d i . 'The ~ likelihood
of migrating ascarids inducing pulmonary eosinophilia in
the principal horses was extremely unlikely given the fact
that these horses were regularly dewormed with ivermectin,
actively training, and housed in racing barns and given al-
most no access to pasture. It is extremely unlikely, there-
fore, that the eosinophil infiltration in the airways of the
horses in the present study was secondary to parasitic lung
disease. We believe that the airway eosinophilia noted in
these horses may be indicative of allergic lung disease, a
situation similar to that previously recognized in the cat'?
and dog.'h
The characterization of this equine disease as solely an
eosinophil-mediated disorder is likely to be erroneous, just
as is the designation of chronic SAD as a purely neutrophil-
mediated disorder. Workers have demonstrated the patho-
physiologic significance of mast cell^,'^,'^ lymphocyte^,?^
and even eosinophilsy,10in the latter condition. It may be
more appropriate to view the principal horses in the present
study as representing part of the natural progression of SAD
given their young age and relatively mild disease state.
The usefulness of the clinical respiratory score described
here in assessing airway disease in horses was addressed in
a previous study? Using other clinical scores, other workers

Table 3. Pulmonary function in principal and control horses in response to exercise challenge. Values are mean 2 SD.
Parameter Baseline 60 Minutes 20 Hours
Rate (breaths per minute) P 13.6 t 4.6 16.9 t- 4.6 15.4 t- 3.1a
C 10.6 t- 1.9 13.5 t 3.2 10.8 t 3.2
Tidal volume (L) P 5.9 t 0.9 5.8 t- 1.2 6.3 t 0.7
C 7.3 t 1.1 6.5 t- 1.0 8.3 t- 2.1
AP,, (cm HZO) P 7.9 t 1.5 8.2 t 1.7 8.8 t 1.3
C 10.7 t 3.3 8.7 t- 3.2 8.7 t 2.9
C,,,, (Lkm H 2 0 ) P 2.9 t- 1.0 2.7 t- 0.5 2.6 t 0.3
C 2.1 t 1.2 2.4 t- 0.7 2.1 t 0.8
R,, (cm HzO/L/second) P 0.72 t 0.16 0.67 t 0.23 0.73 t 0.20
C 0.94 t 0.50 0.70 t 0.34 0.74 t 0.44
WorWminute (kg X m/second/minute) P 380 t 200 450 t 90 450 t 140
C 390 t- 130 450 t 150 300 t 100

P, principal; C, control; AP,,, maximum change in transpulmonary pressure; C,,,, inspiratory dynamic compliance; R, , total lung resistance.
Significantly different from control, P 5 .05.

Pulmonary Eosinophilia in Young Racing Horses
0 5 10 15 20 25
BAL Eosinophil Dflerenthl(%)
Fig 1. Correlation between bronchoalveolar lavage (BAL) eosinophil
differential (%) and respiratory score in study horses; r2 = 0.94, P <
,005.
have demonstrated that semiquantitative measurement of
clinical respiratory signs is a sensitive tool in the diagnosis
of equine lung disease.40Hoffman and coworkers 4 1 further
demonstrated that the discrimination of healthy from dis-
eased foals on the basis of clinical signs alone was a valid
criterion for the definition of distal respiratory tract infec-
tions. The present study further emphasizes the importance
of clinical examination in that it demonstrated a strong cor-
relation between the respiratory score and the degree of
airway eosinophilia.
Upon examination of hematology, fibrinogen, and blood
gases, principal horses were found to differ from controls
in that they had a higher concentration of blood eosinophils.
There was a trend towards higher total leukocyte numbers
that may have been attributable to the peripheral eosino-
philia. It has been suggested that peripheral blood eosino-
phi1 numbers are only a crude indicator of the systemic
state, implying that these cells are in transit from their site
of production to their site of s on sump ti on.^' Indeed, other
workers have found similar increases in blood eosinophil
counts in horses with eosinophilic pulmonary infiltrates.72.33
In the case of the principal horses in the present study, the
assumption is that these peripheral eosinophils were in tran-
sit to the lung. Other body tissues were not, however, ex-
amined in order to test this hypothesis.
The association between Pao, and disease did not
acheive a statistical significance (P = .17). Previous studies
on equine SAD assert that Pao, is a sensitive indicator of
pulmonary function in affected The absence of
prominent differences in PaO, in the face of striking pul-
monary inflammation supports the hypothesis that the hors-
es of the present report were, at the time of study, in re-
mission from their disease. This situation is similar to that
in the chronic heavey horse in clinical remission where
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I67
40
-
I)
-
0
0
I I I I I
M 0 5 10 15 20 25 30
BAL Eosinophil Differential (%)
Fig 2. Correlation between bronchoalveolar lavage (BAL) eosinophil
differential (%) and the provocative concentration of histamine that
caused a 35% decrease in dynamic compliance (PC35C,,,,) (mg/mL
histamine) in study horses; v2 = -0.78, P < 0.05.
BAL neutrophilia may persist in the absence of clinical
signs of airway o b s t r ~ c t i o nAlternatively,
.~~ the level of pul-
monary function alteration in principal horses may not have
been profound enough in these horses to cause impaired
gas exchange at rest, regardless of whether they were in
disease exacerbation or remission.
Results of bronchoalveolar lavage showed that airway
eosinophilia did not occur independently of other cellular
fluxes. Total cell counts in principal horses were approxi-
mately double those in control horses. This increase ap-
peared to be a reflection of the trend towards the harvest
of a greater number of lower airway cells of all types except
epithelial cells in principal horses. Increased cell harvest
was not likely a reflection of BAL technique because as-
piration of the infusate was terminated at 200 mL recovery
in all horses. The increased cell harvest may indicate that
the cells were less adherent to the airways and alveolar
walls in principal horses, that pulmonary eosinophilia is
associated with increases in all inflammatory cells in the
lung, or that an area of higher cell population was selec-
tively lavaged in the principal horses. The latter 2 hypoth-
eses would appear to be most likely in that activated in-
flammatory cells usually are more adherent to respiratory
epithelium than their quiescent counte~parts.4~ Eosinophils
have been demonstrated to produce a variety of cytokines
and chemotactic substances that attract and activate a wide
spectrum of inflammatory cells.27It is well known that la-
vage of larger airways results in higher BAL cell counts
than lavage of the smaller, more distal Given that
the principal horses demonstrated prominent airway re-
sponsiveness compared to controls, it would seem probable
that these horses would be more likely to undergo bron-

168 Hare and Vie1
choconstriction during the BAL procedure, thus resulting
in lavage of somewhat more proximal airways.
When the percentage differentials of the BAL cell con-
stituents were evaluated, it appeared that BAL eosinophilia
was associated with a decrease in the percentage of mac-
rophages. This finding has been observed in horses with
chronic SAD where the prominent neutrophilia results in a
suppression of the macrophage differential.x In both in-
stances, this effect is strictly a result of the dilutional effect
of an increase in cell percentage on the proportion of other
cell types.
The occurrence of increased airway responsiveness in
our principal horses is supportive of the fact that these an-
imals were suffering from SAD, despite the fact that they
did not demonstrate overt airway obstruction. This is in
agreement with the work of Doucet and coworkersZoand
Tesarowski and colleagues,4h who demonstrated the pres-
ence of increased airway responsiveness in horses in clin-
ical remission from SAD. Derksen and colleagues3 con-
cluded that ponies in clinical remission from SAD were not
hyperresponsive to inhaled histamine despite the fact that
their data suggested a trend otherwise. In humans, asth-
matic individuals may retain significant airway hyperreac-
tivity despite the amelioration of their clinical signs with
allergen avoidance or pharmac~therapy.~~ Furthermore, the
real differences observed between principals and controls
with regard to airway responsiveness may have been even
greater than those reported herein. At the time of the his-
tamine challenge (20 hours postexercise) principal horses
had greater respiratory rates and smaller tidal volumes than
did controls. The smaller tidal volumes of the principal
horses could have limited the distribution of the histamine
to the lower airways and may in fact have resulted in higher
than expected values of PC35C,,,.
Airway responsiveness closely correlated with the degree
of airway eosinophilia. This may reflect eosinophil-medi-
ated damage to the airway epithelium or modulation of rest-
ing smooth muscle tone.
In the present study, horses could not be studied after a
period of exacerbation of their SAD with mouldy hay or
straw, a well-known stimulus for this ~ o n d i t i o n . ~This
.~.~
was not possible because the animals were client-owned.
Neither was it possible to achieve consent for lung biopsy
or similar invasive techniques that may have allowed better
characterization of airway pathology.
Given that the clinical signs of respiratory disease were
reported by the trainers to occur in association with exer-
cise, it seemed prudent to examine the animals after a stan-
dardized exercise protocol. Exercise has been studied ex-
tensively as a trigger for both early and late bronchocon-
strictor responses in human asthmatic patients.I2 Although
studies have been performed in the horse documenting the
respiratory response during exercise,4’ to our knowledge,
this was the 1st study to describe the effects of exercise on
the equine respiratory system after exercise challenge.
We exercised the horses of the present study at an inten-
sity that approximated that of racing conditions, insofar as
the clinical signs the horses experienced were associated
with their high level of training. Horses were exacised at
the same time each day to minimize any diurnal influence
on pulmonary function. The treadmill challenge was per-
19391676, 1998, 3, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/j.1939-1676.1998.tb02112.x by Cochrane Colombia, Wiley Online Library on [10/07/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
formed indoors in conditions of controlled temperature and
humidity as these factors have been demonstrated to influ-
ence the occurrence of exercise-induced bronchoconstric-
tion (EIB) in humans.’* In a pilot study, respiratory recov-
ery in both principal and control horses took 20-40 min-
utes, during which time the respiratory rate was markedly
increased from normal. Analysis of pulmonary mechanics
data obtained during this time period could not reliably be
compared with that obtained at rest. For this reason, pul-
monary mechanics measures were not instituted until 60
minutes after treadmill challenge. In humans, EIB is most
severe about 15 minutes after exercise and continues for
approximately 45 minutes.I2 In some individuals, a late-
phase reaction occurs in which bronchoconstriction be-
comes prominent at approximately 4-8 hours and can con-
tinue for 24 hours or more.I2 The protocol we established
did not allow for meaningful interpretation of pulmonary
mechanics until at least 60 minutes after exercise; therefore,
it is possible that acute EIB may have been missed in our
principal horses. Nevertheless, we were not able to dem-
onstrate a difference in airway patency between principal
and control horses either in the early or late phase after
exercise challenge.
The treadmill protocol may have been too demanding for
the purposes of this study. In humans, exercise challenge
for detection of EIB varies greatly but it generally consists
of treadmill running or stationary cycling at a level below
predicted maximal oxygen uptake.12 In our effort to simu-
late racing intensity, we may have exceeded the anaerobic
threshold of our study horses. Plasma lactate concentrations
in 2 horses immediately after cessation of treadmill exercise
were 7.2 and 8.4 mmoVL, respectively, indicating a prom-
inent degree of anaerobic metabolism. Furthermore, all
horses were sweating heavily at the end of the exercise
period. Consequently, it is likely that the hyperpnea ob-
served in all horses in the 30 minutes after exercise was
partly due to thermoregulation. Had the horses been chal-
lenged less vigorously, much of the delay in P F r measure-
ment may have been prevented.
Principal horses had a significantly higher respiratory
rate and reduced tidal volume 20 hours postexercise. In
addition, there was a trend towards reduced tidal volume in
principal horses at rest ( P = .06). The reason for these
differences is not known. However, Doucet4*noted signif-
icantly increased respiratory rate and a trend towards re-
duced tidal volume in young Standardbred horses with ex-
ercise-induced pulmonary hemorrhage (EIPH) when com-
pared to non-EIPH controls. She speculated that part of the
reason for this phenomenon may lie in the fact that these
horses, by virtue of their disease history, may have been
more apprehensive about restraint for diagnostic or thera-
peutic procedures. Regardless of its cause, this observation
may have served to overestimate the dynamic compliance
of the principal horses because this variable is frequency-
dependent and known to decrease with increases in respi-
ratory rate.49
Another reason for our inability to detect EIB may reflect
differences between SAD in horses and asthma in humans.
EIB in humans may occur secondary to the cooling and
rapid rewarming of the bronchi and is a result of impaired
regulation of bronchial vascular tone.’* Pathologic changes

Pulmonary Eosinophilia in Young Racing Horses
in equine SAD are limited to the bronchiolar area: and,
given the likelihood that significant airway cooling does not
occur distal to the conducting airways, temperature f uctu-
ations in this region are unlikely to occur during or after
exercise and EIB, as it occurs in humans, is unlikely to
afflict the horse.
In summary, we have described the phenomenon of
equine SAD associated with airway eosinophilia in the
young performance horse. This condition is associated with
marked airway responsiveness at rest and occurs in the ab-
sence of parasitic infection. Further work in this area is
needed in order to define the extent of disease in affected
horses during clinical exacerbation.
Acknowledgments
We acknowledge Dr David Tesarowski and Mr Dan
Schnurr for their expert technical assistance. This study was
supported by a grant from the Firan Foundation.
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