J Vet Intern Med 2015;29:1728–1734

Thoracic Ultrasonography and Bronchoalveolar Lavage Fluid

Analysis in Holstein Calves with Subclinical Lung Lesions
T.L. Ollivett, J.L. Caswell, D.V. Nydam, T. Duffield, K.E. Leslie, J. Hewson, and D. Kelton
Background: Thoracic ultrasonography (US) and bronchoalveolar lavage fluid (BALF) analysis are antemortem methods
used to identify the lung lesions associated with bovine respiratory disease (BRD). Accuracy of US and the cell distributions
in BALF have not been characterized in calves with subclinical disease.
Objectives: To evaluate the accuracy of US and BALF and describe BALF characteristics in calves with subclinical lung
lesions.
Animals: Twenty-five Holstein calves, 1–12 weeks old.
Methods: In this prospective study, calves with low respiratory scores underwent US, BALF and postmortem examination
(normal US, n = 5; comet-tails, n = 5; consolidation, n = 15). Bronchoalveolar lavage fluid was collected and analyzed for
total and differential cell counts. Lung lesions were assessed by gross and histopathologic examination. Data were analyzed
using nonparametric methods and relative risk analysis. The accuracy of US and BALF were estimated relative to post-
mortem examination.
Results: The sensitivity and specificity of US for detecting lung lesions was 94% (95% CI, 69–100%) and 100% (95% CI,
64–100%), respectively. A cut-point of ≥4% BALF neutrophils was associated with the highest BALF sensitivity and speci-
ficity, 81% (95% CI, 56–94%) and 75% (95% CI, 36–95%). The presence of consolidation on US increased the risk of hav-
ing a BALF neutrophil proportion ≥4% (RR, 3.9; 95% CI, 1.13–13.45; P = .003).
Conclusions and Clinical Importance: Ultrasonography accurately detects lung lesions in calves with subclinical disease.
Clinicians should use a cut-point of ≥4% BALF neutrophils to diagnose subclinical respiratory disease.
Key words: Bovine; Pneumonia; Respiratory; Validation.

linical bovine respiratory disease (BRD) is a

C common cause of morbidity and mortality in
young dairy heifers.1 Within-herd prevalence is highly
Abbreviations:
BAL bronchoalveolar lavage
variable, ranging from 0 to 90% of calves affected.2,3 BALF bronchoalveolar lavage fluid
This within-herd variation could result from differences BRD bovine respiratory disease
in disease frequency, recording method, or scoring ICS intercostal space
system. The constituents of scoring systems vary, but in LS ultrasonographic lesion score
general they are based on the combined severity of RS respiratory score
sBRD subclinical bovine respiratory disease
Se sensitivity
Sp specificity
From the Department of Medical Sciences, UW-Madison School TNCC total nucleated cell count
of Veterinary Medicine, Madison, WI (Ollivett); the Department of US thoracic ultrasonography
Pathobiology, Ontario Veterinary College, University of Guelph,
Guelph, Ontario, Canada (Caswell); the Department of Population
Medicine and Diagnostic Sciences, Cornell University College of
Veterinary Medicine, Ithaca, NY (Nydam); the Department of several clinical signs associated with respiratory
Population Medicine, Ontario Veterinary College, (Duffield, Leslie, disease.4,5 Pitfalls of scoring systems include the subjec-
Kelton); and the Department of Clinical Studies, Ontario Veterinary tive nature of ranking the severity of clinical signs5 as
College, University of Guelph, Guelph, Ontario Canada (Hewson). well as the inherent failure to identify calves with sub-
This work was performed at the Elora and Ponsonby Dairy clinical bovine respiratory disease (sBRD).
Research Centres, University of Guelph, Ontario, Canada.
This study was presented as a research abstract at the 2013 The association between postmortem lung lesions and
American Association of Bovine Practitioners Annual Conference, decreased average daily gain in the absence of clinical
Milwaukee, WI (by Ollivett) and 2014 American College of Veteri- signs is evidence of sBRD in beef and veal calves.6,7
nary Internal Medicine Annual Forum, Nashville, TN (by Hewson). Others have documented increased neutrophil propor-
This manuscript was part of the PhD thesis submitted by Dr. tions8,9 or pathogens in bronchoalveolar lavage fluid
Theresa Ollivett to the University of Guelph. (BALF) from clinically normal calves.10 Although little
Corresponding author: T.L. Ollivett, Department of Medical
Sciences, UW-Madison School of Veterinary Medicine, Madison,
work has been done to define and measure sBRD in dairy
WI 53706; e-mail: [email protected]. calves, the association between greater body weight gains
Submitted March 2, 2015; Revised May 25, 2015; Accepted after implementation of metaphylaxis suggests the pres-
July 28, 2015. ence of sBRD.11 Accurate antemortem methods of
Copyright © 2015 The Authors. Journal of Veterinary Internal detecting sBRD by identification of lung lesions or detec-
Medicine published by Wiley Periodicals, Inc. on behalf of American tion of low-grade lung inflammation will improve classifi-
College of Veterinary Internal Medicine. cation of disease in both individuals and herds.
This is an open access article under the terms of the Creative
Commons Attribution-NonCommercial License, which permits use,
Direct methods of identifying lung inflammation have
distribution and reproduction in any medium, provided the original relied on sampling the airways by means of transtra-
work is properly cited and is not used for commercial purposes. cheal aspiration or bronchoalveolar lavage (BAL). Both
DOI: 10.1111/jvim.13605 methods are invasive, seldom used on the farm, and
 19391676, 2015, 6, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/jvim.13605 by Readcube (Labtiva Inc.), Wiley Online Library on [26/12/2024]. 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
Subclinical Lung Lesions in Dairy Calves 1729

cut-points for differential cell counts have not been well applied to the hair as the transducing agent. The hair was not
defined. Indirect methods of detecting lung inflamma- shaved. Systematic scanning started at the level of the epaxial mus-
tion identify the downstream effects of inflammation, cles in the right/left 10th intercostal space (ICS). Within each ICS,
such as fever and acute phase proteins,12 but these tests the probe was positioned parallel to the ribs and moved ventrally
toward the costal arch or the sternum until specified ultrasono-
lack specificity (Sp).
graphic landmarks were visualized (Tables 1, 2). The probe was
Recently, interest in using thoracic ultrasonography moved cranially to examine each ICS up to the right 1st or left
(US) to diagnose the lung lesions associated with sBRD 2nd ICS (Fig 1). The scapula marked the dorsal margin of the
in dairy calves has grown.13–15 Bacterial and occasion- examination cranial to the 7th ICS bilaterally. The lung adjacent
ally viral respiratory diseases result in non-aerated to the right 4th through 1st ICS and left 4th through 2nd ICS was
superficial lung lobules. This changes lung density, alter- examined with the transducer between the forelimb and the cranial
ing the US image from that of a strong reflector with ventral thoracic body wall. Peripheral lung tissue was considered
reverberation artifact to a homogenous hypoechoic normal when a hyperechoic line with reverberation artifact was
structure similar to liver.16 present signifying the interface between the high impedance tissue
The accuracy of US has been documented in clinical of the thorax and the low impedance tissue of the lung.21 Pleural
roughening, or comet-tailing artifact, was noted when a vertical
cases of bronchopneumonia,17,18 but few of these
hyperechoic line emanated from the pleural surface.21 Lung lesions
reports were prospective or case-controlled19,20 and (also referred to as consolidated lung or nonaerated lung)
none examined sBRD. Therefore, the first objective of appeared hypoechoic and lacked both the bright white band at the
this study was to determine the sensitivity (Se) and Sp
of a widely available, portable US unita in detecting the
lung lesions associated with sBRD in apparently healthy Table 2. Landmarks for the left lung during ultrasono-
calves. The second objective was to evaluate BALF graphic examination.
characteristics and develop a BALF neutrophil propor-
tion cut-point for determining Se and Sp for detecting Lung Lobe
sBRD. We hypothesized that US would accurately Caudal Aspect Cranial Aspect
detect lung lesions and that these lesions would be asso- Caudal of Cranial Lobe of Cranial Lobe
ciated with inflammatory BALF.
L – ICS 6–10 4–5 2–3
Ventral Diaphragm CCJ & pleural Heart
Materials and Methods landmark(s) deviation

General L – ICS, left intercostal space; CCJ, costochondral junction.

This prospective study was completed between January 1, 2012
and December 15, 2012 at the Elora and Ponsonby Dairy Research
Centres of the University of Guelph in southwestern Ontario,
Canada. Sixty-two 3–6-day-old Holstein bull calves were enrolled
into a separate study evaluating the effect of an intranasal respira-
tory vaccine over a 12-week follow-up period. From this popula-
tion, 25 calves were selected for the current intensive study. Calves
were raised in individual stalls, fed whole milk until 6 weeks of age,
and moved to groups by 8 weeks. The Animal Care Committee of
the University of Guelph approved this study (AUP #11R110).

Respiratory Scoring and Thoracic Ultrasonography

As part of the other study, one author (TO) and a research
technician visited the research centers twice weekly to determine
respiratory scores (RSs) according to a standardized respiratory
scoring system.4 Calves with RS > 4 were considered sick4 and
were excluded from the current intensive study.
Once per week, immediately after the RS, US was performed
using a portable linear rectal ultrasound unita, set at a depth of Fig 1. Right lung in situ. The lung is outlined in white. Ribs are
9 cm, frequency of 6.2 MHz, and gain of 16 dB (near 13 dB far labelled by number. Cranial aspect of the right cranial lobe is situ-
36 dB). Approximately 300 mL of 70% isopropyl alcohol was ated in the 1st and 2nd intercostal spaces.

Table 1. Landmarks for the right lung during ultrasonographic examination.

Lung Lobe

Caudal Middle Caudal Aspect of Cranial Lobe Cranial Aspect of Cranial Lobe
R-ICS 6–10 5 3–4 1–2
Ventral landmark(s) Diaphragm CCJ & pleural deviation Heart Internal thoracic artery & vein

R-ICS, right intercostal space; CCJ, costochondral junction.

 19391676, 2015, 6, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/jvim.13605 by Readcube (Labtiva Inc.), Wiley Online Library on [26/12/2024]. 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
1730 Ollivett et al

pleural interface and reverberation artifact.16 Lung lesions were tissues, and rib caudal to the lesion were reflected dorsally to
documented according to location and size as measured by the confirm location of the lesion with respect to external landmarks
ventral-dorsal distance. All observations were dictated, digitally (Fig 2). Once confirmed, all of the ribs were reflected dorsally to
recorded, and later transcribed to a database. expose the whole affected lung. The heart and lung were removed
en bloc, placed on ice, and transferred back to the University
pathology suite for further evaluation.
Animal Selection for Intensive Study
Once in the pathology suite, all lung lesions were recorded on a
Selection criteria for this intensive study included: RS < 5, pre- standard diagram after thorough palpation of the tissues to con-
viously normal US, and no antimicrobial treatment within the firm consolidation and identify any pulmonary changes within the
prior 2 weeks or since birth if calves were <2 weeks old. Based on parenchyma. Tissue samples for histopathology were taken from
sample size calculations, 4 calves were needed per comparison the border zone between normal and consolidated lung. Additional
group to detect a difference of 10% in the BALF neutrophil pro- samples for histopathology were taken from the right and left cra-
portion (standard deviation, 5%; power, 80%; alpha = 0.05). nial lobes, the right middle lung lobe, and the right and left caudal
Therefore, for this study, the goal was to obtain 5 calves for each lobes in unaffected portions of affected lungs. In unaffected lungs,
category of a 5-point ultrasonographic lesion score (LS) for a total samples for histopathology were taken from the right and left
of 25 calves. The categorizations were based on one author’s clini- cranial lobes, the right middle lung lobe, and the right and left
cal experience (TO) regarding normal (LS = 0), very mild caudal lobes. Routine hematoxylin and eosin stains were made of
(LS = 1), mild (LS = 2), moderate (LS = 3), and severe (LS = 4)
levels of ultrasonographic lung lesions in large animals at a
University hospital. Definitions for each were as follows: LS0: nor- A
mal aerated lung imaged as a smooth hyperechoic line adjacent to
the body wall with reverberation artifact and no comet-tail artifact
or hypoechoic consolidations; LS1: comet-tail artifacts imaged as
vertical hyperechoic lines emanating from the pleural surface with-
out hypoechoic consolidations; LS2: <1 cm of hypoechoic consoli-
dation lacking the hyperechoic line of the pleura and reverberation
artifact in the area of the lesion; LS3: 1–3 cm of hypoechoic con-
solidation; and LS4: >3 cm of hypoechoic consolidation. Consoli-
dations were measured in the dorsal-ventral plane.
Approximately once a week, 1–2 calves that met the above cri-
teria were selected for BALF collection, euthanasia, and post-
mortem examination until all 25 calves were obtained.

Collection and Analysis of Bronchoalveolar

Lavage Fluid
Calves were sedated with xylazine hydrochlorideb (0.05 mg/kg
IV) and butorphanolc (0.1 mg/kg IV) and restrained in sternal
recumbency. The external naris was cleaned with gauze and a
9 mm (outer diameter), 1.5 m flexible fibreoptic bronchoscoped
previously sterilized with a 2% glutaraldehyde solution, was intro-
duced into the ventral nasal meatus of the clean naris. The bron-
choscope then was passed through the nasopharynx into the
trachea, extending into a distal airway within the lung containing
the ultrasonographic evidence of consolidation or, if normal, the
right lung. Once wedged into the bronchus, 2 aliquots of 120 mL
sterile saline4 were sequentially dispensed into and retrieved from
the airway via the biopsy channel. Once 50% of the aliquot was
retrieved, the scope was removed. Diagnostic samples were taken
from the last aliquot retrieved, placed in Ca-EDTA commercial B
blood collection tubes, and held in an ice water bath until submis-
sion to the Animal Health Laboratory at the University of
Guelph. Samples were evaluated within 4 hours of collection.
Wright-stained sediment and cytocentrifuge preparations of BALF
were evaluated to determine the 200 cell differential cell count.
Automated methodse were used to determine total nucleated cell
count (TNCC).

Euthanasia and Postmortem Examination

Fig 2. Right lung with lobar consolidation (LS4) of the cranial
Immediately after BALF collection, calves were euthanized aspect of the right cranial lobe. (A) In situ specimen. Carcass is in
using captive bolt according to the AVMA’s Guidelines for left lateral recumbency. The right 2nd rib (white star) is reflected
Euthanasia. A respiratory system-based postmortem examination dorsally exposing the combined 1st and 2nd intercostal space,
was performed immediately after euthanasia by one author (TO). revealing the cranial aspect of the right cranial lobe. (B) Right
During the postmortem examination, the carcass was in lateral lung removed, showing consolidation of cranial aspect of cranial
recumbency with the affected lung up. The skin, subcutaneous lobe (white star).
 19391676, 2015, 6, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/jvim.13605 by Readcube (Labtiva Inc.), Wiley Online Library on [26/12/2024]. 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
Subclinical Lung Lesions in Dairy Calves 1731

prepared histopathology sections after at least 24 hours of fixation Results

in 10% buffered formalin. The histologic appearance of the lung
was categorized as either normal or consistent with bacterial pneu- One BALF sample was damaged in transport, result-
monia (ie, the presence of neutrophils within the bronchiolar or ing in 24 calves for analysis (LS0, n = 4; LS1, n = 5;
alveolar lumens) or viral pneumonia (ie, the presence of mononu- LS2, n = 5; LS3, n = 5; LS4, n = 5). Descriptive statis-
clear peribronchial or peribronchiolar infiltrates, with or without tics for age, RS, US lesion size, and BALF for each US
epithelial necrosis and lobular atelectasis). category are summarized in Table 3.
The Se and Sp of US for detecting the lung lesions
Statistical Analyses associated with sBRD was 94% (95% CI, 69–100%)
Medians and interquartile ranges were calculated for the follow-
and 100% (95% CI, 64–100%), respectively. Ultrasono-
ing variables: age, RS, US lesion size, TNCC, and differential cell graphic consolidation was associated with firm, red lung
counts including neutrophil proportion, macrophage proportion lesions on gross examination in all cases (n = 15). All
and lymphocyte proportion. For the purposes of analyses, in addi- lung lesions were located within the cranial or right
tion to describing each US category individually, all calves with middle lung lobes. The cranial aspect of the right cra-
US consolidation (LS2–4) were combined into 1 group (consoli- nial lung lobe was completely consolidated in 4/5 LS4
dated, n = 15) for comparison to completely normal (LS0, n = 4) calves (Figs 2, 3). Ultrasound examination failed to
calves and calves with comet-tailing (LS1, n = 5). The TNCC, neu- detect a 1 cm area of atypical consolidation located in
trophil proportion, macrophage proportion and lymphocyte pro- the dorsomedial aspect of the right lung of 1 calf, but
portions were compared using Wilcoxon rank sum test.f
severe diffuse comet-tailing artifacts were observed
Sensitivity and Sp were estimated for US using gross post-
mortem examination as the gold standard. Ultrasonographic lung
within the right 5th ICS of this particular calf.
lesions (positive = any nonaerated lung imaged; negative = only Histopathology results are summarized in Figure 4.
aerated lung imaged) and gross postmortem lesions (posi- Briefly, 4/4 LS0 calves had normal histopathology
tive = presence of dark red, firm lung tissue; negative = absence results, whereas 5/5 LS4 calves had evidence of only
of dark red, firm lung tissue) were coded as dichotomous results. bacterial lung lesions. Both viral and bacterial lesions
Sensitivity was calculated by dividing the number of calves were present histologically in calves in the remaining
affected with both US and gross lesions (true positives) by the categories (LS1–3).
total number of calves with gross lesions (true positive + false The TNCC was 0.52 9 109 cells/L in BALF from
negatives). Specificity was calculated by dividing the number of consolidated lungs (LS2–4, n = 15) versus
calves without either US or gross lesions (true negatives) by the
0.59 9 109 cells/L from completely normal lungs (LS0,
total number of calves without gross lesion (true negatives + false
positives). Exact confidence intervals (95%) were calculated for
n = 4; P = .65). The neutrophil proportion was 14% in
both Se and Sp. BALF from consolidated lungs versus 1.25% from com-
A receiver operator characteristic (ROC) curve was developed pletely normal lungs (P = .005). The neutrophil propor-
to determine which BALF neutrophil proportion provided the tion in calves with comet-tailing (LS1, n = 5) was 2%
highest combined Se and Sp for predicting lung lesions using gross versus 14% from consolidated lungs (P = .05) and
postmortem examination as the gold standard. Exact confidence 1.25% from completely normal lungs (P = .22). The
intervals (95%) were calculated for both Se and Sp. The relative BALF macrophage proportion was 79% from consoli-
risk (95% CI) was calculated based on a contingency table for the dated and 97% from completely normal lungs
outcome, BALF neutrophil proportion ≥4%, and the predictor, (P = .006). The macrophage proportion in calves with
US lung consolidation (LS2–4). Alpha was set at ≤.05 except dur-
comet-tailing was 91% compared to 97% from that of
ing multiple comparisons in which Bonferroni’s correction for 5
comparisons set alpha at ≤.01). Unless otherwise noted, commer-
completely normal lungs (P = .06) and 79% from con-
cially available software was used for analysis.g solidated lungs (P = .24).

Table 3. Description of respiratory score (RS) and bronchoalveolar lavage fluid (BALF) findings grouped by ultra-
sonographic (US) lesion score (LS) in Holstein dairy calves. All values represent medians (interquartile ranges).
Varying Levels of US Consolidation
Normal Comet-Tails All US Consolidation
Variable LS0, n = 4 LS1, n = 5 LS2, n = 5 LS3, n = 5 LS4, n = 5 n = 15
Age (days) 28 (25–41) 32 (25–65) 68 (54–76) 62 (24–64) 71 (71–84) 65 (52–74)
RS 2 (2–2) 2 (2–3) 2 (2–4) 3 (3–3) 2 (2–3) 3 (2–4)
US lesion (cm) 0 0 0.5 (0.25–0.50) 1.0 (1.0–1.5)b 6 (4–8)b 1.5 (0.63–4)a,b
TNCC 0.59 (0.45–0.80) 0.34 (0.30–0.90) 0.51 (0.25–0.73) 0.46 (0.33–0.56) 0.72 (0.52–0.82) 0.52 (0.29–0.73)
(9109 cells/L)
Neut (%) 1 (1–2) 2 (2–11) 25 (9–28) 8 (3–14) 17 (12–20) 14 (7–22)a
Mac (%) 97 (96–99) 91 (78–93) 69 (67–79) 82 (80–92) 79 (74–85) 79 (74–89)a
Lymph (%) 1 (1–2) 7 (5–8) 6 (3–11) 5 (4–8) 3 (3–3) 4 (3–8)

LS0, completely normal ultrasonographic exam; LS1, comet-tailing artifacts on ultrasonographic exam; LS2, <1 cm consolidation on
ultrasonographic exam; LS3, 1–3 cm consolidation on ultrasonographic exam; LS4, >3 cm consolidation on ultrasonographic exam;
TNCC, total nucleated cell count; Neut, BALF neutrophil proportion; Mac, BALF macrophage proportion; Lymph, BALF lymphocyte
proportion.
a
Differs significantly from LS0. bDiffers significantly from LS1. Bonferroni’s correction for multiple comparisons P ≤ .01.
 19391676, 2015, 6, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/jvim.13605 by Readcube (Labtiva Inc.), Wiley Online Library on [26/12/2024]. 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
1732 Ollivett et al

Fig 5. Receiver operator characteristic curve demonstrating the

neutrophil proportion ≥4% in bronchoalveolar lavage fluid pro-
vides the highest combined sensitivity and specificity for detecting
subclinical lung lesions in Holstein dairy calves (n = 24).

in dairy calves with sBRD. Identification of US lung lesions

accurately predicted the presence of gross and histopatho-
logic lung lesions and was associated with an increased
proportion of BALF neutrophils.
Fig 3. Ultrasonographic image of consolidated lung imaged from Other work has supported the general accuracy of
the right 1st intercostal space. Image orientation: left = dorsal, US,17,19,20 but direct comparisons among studies should
right = ventral, top = superficial, bottom = deep. The 2 blood ves- be made with caution considering differences in tech-
sels are the internal thoracic artery and vein.
nique, equipment, and study design. Previously,
researchers did not routinely and systematically extend
their examinations cranially beyond the 3rd ICS.17–20,22–24
In the current study, 4 of 5 of the most severely affected
calves would have been misclassified as normal had
these previously mentioned techniques been imple-
mented because the lesions were visible only from the
right 1st and 2nd ICS. Because BRD often starts in the
cranial aspect of the right cranial lung lobe, it is logical
that this lobe must be imaged during examination.25,26
Although older studies often used linear probes, none
report using a linear rectal transducer. The streamlined
design of the rectal transducer is conducive to a better
examination of the cranial thorax, specifically the right
1st and 2nd ICS. The handle and cord extend from the
Fig 4. Lung tissue histopathology results (n = 24). Black = no midsection of traditional probes instead of the end, pre-
lesions. Gray = bacterial lesion. Striped = viral lesion. LS0: no venting it from easily sliding between the forelimb and
ultrasound (US) consolidation; LS1: comet-tail artifact; LS2: the cranial thoracic body wall for evaluation of the cra-
<1 cm US consolidation; LS3: 1–3 cm US consolidation; LS4: nial aspect of the cranial lobes. This area previously
>3 cm US consolidation. was thought to be unreachable.18
In the current study, 1 case of atypical consolidation
The highest BALF Se (81%, 95% CI, 56–94%) and was not detected because it was surrounded by aerated
Sp (75%, 95% CI, 36–95%) was associated with a cut- lung. Another study17 observed a similar situation in
off of ≥4% neutrophils based on the receiver operator which a 10 cm abscess was obscured by aerated lung.
characteristics (AUC, 0.85; 95% CI, 0.67–1.0; Fig 5). Ultrasound waves cannot penetrate aerated lung tissue,
The presence of US lung lesions increased the risk of which serves to hide lesions deep within the parench-
having a BALF neutrophil proportion ≥4% (RR, 3.9; yma. Fortunately, most lung lesions associated with
95% CI, 1.13–13.45; P = .003). BRD extend to the pleural surface.25,26
Analysis of BALF is used to detect the pulmonary
Discussion inflammation associated with BRD and has been used to
control the confounding effect of sBRD.8 Suggested cut-
To the authors’ knowledge, this is the first study points for the proportion of BALF neutrophils are vari-
evaluating a defined US technique and BALF characteristics able, ranging from 10 to 40%.4,8 Normal calves in the
 19391676, 2015, 6, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/jvim.13605 by Readcube (Labtiva Inc.), Wiley Online Library on [26/12/2024]. 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
Subclinical Lung Lesions in Dairy Calves 1733

current study had neutrophil proportions similar to those Chung, and Sarah Stanger-Guy as well as the Elora
in another study27 which sampled normal lungs at and Ponsonby Dairy Research Centre and Animal
necropsy. Neutrophil proportions also were similar to a Health Laboratory staff.
previous report8, but 18 of 30 clinically normal calves Conflict of Interest Declaration: Authors disclose no
were excluded from that study8 for having neutrophil conflict of interest.
proportions >10%. Others have reported much higher Off-label Antimicrobial Declaration: Authors declare
BALF neutrophil proportions in clinically normal con- no off-label use of antimicrobials.
trol animals,9,10 likely reflecting the presence of sBRD in
the control population. Results from the current study References
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E-22VGS99x17 Veterinary Fiberscope, LSVP International, Los 13. Buczinski S, Forte G, Francoz D, B
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Stata 12.1, Stata Corp LP, College Station, TX 14. Buczinski S, Fort
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