Avian Pathology

ISSN: 0307-9457 (Print) 1465-3338 (Online) Journal homepage: https://www.tandfonline.com/loi/cavp20

Temporal Characterization of Wooden Breast

Myopathy (“Woody Breast”) Severity and
Correlation with Growth Rate and Lymphocytic
Phlebitis in Three Commercial Broiler Strains and a
Random-Bred Broiler Strain

Laura R. Chen, M. Mitsu Suyemoto, Albarra H. Sarsour, H. Alejandro

Cordova, Edgar O. Oviedo-Rondón, Michael Wineland, H. John Barnes & Luke
B. Borst

To cite this article: Laura R. Chen, M. Mitsu Suyemoto, Albarra H. Sarsour, H. Alejandro Cordova,
Edgar O. Oviedo-Rondón, Michael Wineland, H. John Barnes & Luke B. Borst (2019): Temporal
Characterization of Wooden Breast Myopathy (“Woody Breast”) Severity and Correlation with
Growth Rate and Lymphocytic Phlebitis in Three Commercial Broiler Strains and a Random-Bred
Broiler Strain, Avian Pathology, DOI: 10.1080/03079457.2019.1598541

To link to this article: https://doi.org/10.1080/03079457.2019.1598541

Accepted author version posted online: 27

Mar 2019.

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Publisher: Taylor & Francis & Houghton Trust Ltd

Journal: Avian Pathology
DOI: 10.1080/03079457.2019.1598541

Temporal Characterization of Wooden Breast Myopathy (“Woody Breast”) Severity and

Correlation with Growth Rate and Lymphocytic Phlebitis in Three Commercial Broiler Strains

and a Random-Bred Broiler Strain

Laura R. Chena, M. Mitsu Suyemotoa, Albarra H. Sarsourb, H. Alejandro Cordovab, Edgar O. Oviedo-Rondónb,
Michael Winelandc, H. John Barnesa, Luke B. Borsta*
a
Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State
University, 1060 William Moore Drive, Raleigh, NC 27607
b
Prestage Department of Poultry Science, North Carolina State University, Box 7608 Raleigh, NC, 27695
c
Hatchery Consult LLC, 226 West Young Street, Rolesville, NC 27571
*Email address of corresponding author: lbborst@ncsu.edu
Abstract
Wooden breast myopathy (WBM), or “woody” or “wooden” breast affects modern, rapidly growing, high breast-
yield broiler chickens. Decreased meat quality due to undesirable organoleptic properties and condemnation of
affected breast meat cause economic losses. The pathogenesis of WBM remains unknown. In this study, WBM
lesion development was determined in 3 modern broiler strains and Athens Canadian Random Bred (ACRB)
broilers, a 1950’s unselected broiler chicken. Correlations between WBM severity and incubation temperature
profile, sex, strain, body weight, and lymphocytic phlebitis were also determined. At 2, 4, 6, and 8 weeks, samples
of breast muscle from 10 male and 10 female birds from each strain, incubated under optimal or low-early, high-late
temperatures, were scored histologically for severity of WBM and lymphocytic phlebitis. WBM lesions, identified
as early as 2 weeks, became progressively more severe with age and growth in the three commercial broiler strains.
WBM severity was significantly correlated with lymphocytic phlebitis and body weight. Lymphocytic phlebitis and
minimal WBM were present in the ACRB broilers at all samplings, but did not progress in severity over time. There
were no significant differences in severity of WBM among the commercial broiler strains, between sexes, or
between incubation temperature profiles. The positive correlation between WBM severity and lymphocytic phlebitis
indicate vascular injury is likely an important factor in the pathogenesis. Mild muscle lesions in ACRB birds without
overt clinical signs indicate subclinical muscle disease may have been present in broilers prior to the description of
woody breast myopathy.
Key Words: Athens Canadian Random Bred, broiler chickens, growth rate, incubation temperature, lymphocytic
phlebitis, Pectoralis major muscle, woody breast myopathy
Introduction
Wooden breast myopathy (WBM), also known as “woody” or “wooden breast” in its most severe form, is a

myopathy primarily affecting the superficial breast muscle (pectoralis major) that has recently emerged in modern

rapid growth, high-breast-yield broiler chickens (Sihvo et al., 2014). Despite its relatively short recognition in the

poultry industry, woody breast has a worldwide distribution (Sihvo et al., 2014; Mutryn et al., 2015; de Brot et al.,

2016) and results in significant economic losses due to downgrading of meat quality (Kuttappan et al., 2012;

Petracci et al., 2015; Trocino et al., 2015). Recent disposition instructions from the United States Department of

Agriculture Food Safety Inspection Service will likely increase its economic impact in the near future (USDA FSIS

2018).
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Over the growing period, affected chickens do not exhibit clinical signs (Sihvo et al., 2014; de Brot et al., 2016,

Sihvo et al., 2017). Lesions were infrequently identified in organs other than the pectoralis major muscle when

affected chickens were examined at processing or early in the growing period in experimental studies (Sihvo et al.,

2014; Trocino et al., 2015; de Brot et al., 2016). Woody breast is characterised by pallor, white striations, and

strictured and coalescing areas of fibrosis, predominantly in the cranial portion of the pectoralis major muscle

(Sihvo et al 2014; Clark and Velleman 2016; Sihvo et al 2017). In more severely affected birds, the overlying fascia

can also be variably expanded by gelatinous fluid and occasional haemorrhage (Sihvo et al., 2014). Histologically,

gross changes correlate with polyphasic, segmental, myodegeneration and regeneration accompanied by a mixed

inflammatory response and, interestingly, a robust lymphocytic phlebitis (Sihvo et al 2014; de Brot et al 2016, Papah

et al., 2017; Sihvo et al 2017). Structural alterations in myofibers correlate with altered expression of genes involved

in reacting to an injurious and subsequently regenerative stimulus, including reactive oxygen species metabolism,

oxidative stress and signal transduction, muscle development, and blood vessel morphogenesis, among others

(Mutryn et al., 2015; Velleman & Clark 2015; Abasht et al., 2016; Clark & Velleman 2016; Zambonelli et al.,

2016).

While the structural and functional alterations of affected pectoralis major muscles have been described, the

underlying pathogenic pathway remains incompletely understood with a complex, multifactorial process proposed

(Zambonelli et al. 2016). Increased body weight and growth rate have been among the most extensively discussed

factors, with it now thought that they have at the very least a contributory role (Sihvo et al., 2014; Velleman and

Clark 2015; de Brot et al 2016). There is a myopathy in poultry species associated with vitamin E and selenium

deficiency (Kuttapan et a., 2016), but dietary trace mineral levels, including selenium, have been examined without

evidence that they contribute to WBM prevalence or severity (Meluzzi et al. 2016; Sihvo et al. 2017). Rauber et al.

(2016) examined the role of dietary lysine in WBM severity and found a positive correlation, but within the context

of a concurrent increased growth rate. Incubation temperature conditions influence myogenesis and meat quality

traits (Janisch et al., 2015), but have not been examined within the context of WBM. Genotype predispositions for

WBM have been examined amongst modern broiler strains by multiple researchers but without evidence of a

correlation (Bailey et al 2015; Trocino et al., 2015).

A frequently observed microscopic lesion within affected pectoralis major muscles is a robust lymphocytic phlebitis

that has been noted by multiple previous researchers (Sihvo et al. 2014, Sihvo et al. 2017, Papah et al., 2017), with a
 3

recent publication identifying the early presence of this lesion and its potential contributory role to WBM (Papah et

al., 2017).

Further elucidation of the pathogenesis of WBM was the broad goal of this research, with specific objectives to

characterize temporal patterns of lesion development and evaluate correlations between WBM severity and

incubation temperature profile, sex, strain, body weight, and lymphocytic phlebitis in an experimental setting. Three

contemporary rapid-growth, high-breast yield commercial broiler strains were compared with ACRB broilers, a

random-bred strain unselected since the 1950’s (Collins et al., 2016).

Materials and methods

Experimental design
Experimental use of birds was approved by the Animal Care and Use Committee and conducted in compliance with

the Guidelines for Care and Use of Laboratory Animals at North Carolina State University.

Experimental design is as previously described (Chen et al., 2017). Briefly, 8 treatments that included 4 broiler

chicken strains and 2 incubation temperature profiles were evaluated for severity of WBM and lymphocytic phlebitis

in the pectoralis major muscle at 2, 4, 6, and 8 weeks of age. Chicken strains included 3 modern commercial

chicken strains (A/A, A/B, and C/C) and the Athens Canadian Random Bred (ACRB) chicken, a line representative

of 1950’s broiler chickens, obtained from the University of Georgia (Athens, GA) (Collins et al., 2016).

Fertile eggs from all strains were randomly divided between two incubators programmed with either standard (S) or

low-high (LH) incubation temperature profiles as previously described (Da Costa et al., 2016). The S temperature

profile maintained optimal eggshell temperature at 37.8C in the incubator by modulating ambient temperature. In

the hatcher for treatment S, ambient temperature was initially set at 36.9C, decreasing to maintain an eggshell

temperature no greater than 37.9C and then decreasing slowly to 34.7C at the time chicks were removed. For the

LH temperature profile, ambient temperature was set to 37.4C in both the incubator and hatcher without

modulation based on eggshell temperature, resulting in eggshell temperatures below optimum during the first 72

hours of incubation and eggshell temperatures higher than optimum following the 16 th day of incubation.

Temperatures were decreased for the final 24 hours in the hatcher to maintain hatchability.

On the day of hatch, chicks were weighed, sex-identified, individually tagged, and placed into 80 floor pens with 12

birds per pen (n  =  960). There were five pen replicates (60 birds) for each broiler line/sex/incubation profile
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combination. Birds were raised on 6 inches of pine shaving litter covering concrete floors. Birds were fed ad libitum

with diets formulated to meet nutritional requirements containing corn, soybean meal, and distiller’s dried grains

with solubles, milled into a crumble for the starter diet (days 0–14) and pellets for the grower (days 15–35) and

finisher (days 36 to end of study) diets. Nicarbazin (1 lb/ton) was added to the diets to control coccidia.

Sample collection
Two birds per pen were randomly sampled at 2, 4, 6, and 8 weeks of age for a total of 160 birds at each sampling

time point and 10 birds per broiler line/sex/incubation temperature combination. Due to poor hatching rates and

sporadic mortality, particularly in the ACRB, fewer than 10 birds per combination were collected at early time

points to ensure sufficient birds were available at the end of the study. At each time point, birds were euthanized by

cervical dislocation and weighed. A 1-2 cm thick section of the pectoralis major muscle extending from the left

coracosternal (shoulder) joint to the cranial keel at the thickest part of the muscle was collected from each bird,

refrigerated and kept moist with normal saline (0.9% NaCl) until the passage of rigor mortis. The tissue was then

consistently trimmed to provide a full thickness block one cm wide and placed into 10% neutral buffered formalin.

Tissue processing
Two faces of each formalin-fixed muscle sample were removed to obtain both cross and longitudinal sections of

myocytes. Tissues were processed into paraffin wax blocks, cut into 3 to 5 µm thick sections, mounted on glass

slides, and stained with haematoxylin and eosin.

Histomorphologic scoring
Severity of WBM and severity and distribution of lymphocytic phlebitis were both scored in one tissue cross

section. Scoring was performed by one author (LRC) blinded to the sex, strain, and incubation temperature

conditions. A random subset of tissue cross sections was subsequently reviewed by a second author (HJB). WBM

was scored based on the percentage of myocytes undergoing degeneration, necrosis, or regeneration in 10

consecutive 200X (surface area 0.950 mm2) magnification fields as follows: 1  =  0% myocytes affected; 2  =  1–33%

myocytes affected; 3  =  34–66% myocytes affected; and 4  =  67–100% myocytes affected. As WBM lesions are not

uniform throughout the muscle, the recorded score represented the most severely affected area in the tissue section.

Typical lesions representing the different histomorphologic scores are shown in Figure 1.
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Lymphocytic phlebitis was scored in the same tissue section as above based on the number and distribution of

lymphocytes around or in vein walls across the entire section as follows: 1  =  no lymphocytes; 2  =  lymphocytes

infiltrating less than 5 cells thick, focal to multifocal; 3  =  lymphocytes infiltrating less than 5 cells thick, multifocal

and greater than 5 cells thick, focal; and 4  =  lymphocytes greater than 5 cells infiltrating, multifocal (Figure 2).

Interstitial edema, fibrosis and histiocytic and heterophilic inflammation were interpreted as reactive processes and

were not included in either scoring system.

A total of 608 sections were scored for both WBM and lymphocytic phlebitis.

Statistical analyses
Mean WBM scores including all time points and by week were compared among broiler strains and between

incubation temperature profiles and sex using the non-parametric Kruskal-Wallis test with pairwise post-hoc

comparisons performed using a Mann-Whitney U test with Bonferroni’s correction on the p-value. Incubation

temperature profiles and sex were compared within a strain. Mean WBM scores by strain were compared among

weeks using the same methodology. Mean body weights by sex and strain were compared among broiler strains at

weeks 2, 4, 6, and 8 using a one-way analysis of variance (ANOVA) with pairwise post-hoc comparisons using the

Tukey-Kramer method. Correlation of body weight to WBM score; time to WBM score; time to lymphocytic

phlebitis score; and lymphocytic phlebitis score to WBM score was determined by calculating Spearman’s

correlation coefficient rho (rs), with calculations performed by week when body weight was one of the variables. In

all cases, significance was established to be p < 0.05. JMP Pro 13 (Cary, North Carolina) was used for all statistical

analyses.

Results
Histologic features of WBM
Fulminant WBM lesions in the pectoralis major muscle were characterized by multifocal, marked, polyphasic

myocyte degeneration and necrosis with loss of sarcoplasmic cross striations, sarcoplasmic swelling and

fragmentation, vacuolation, fiber splitting, fiber atrophy, contraction bands, granular mineralization, and pyknosis to

karyolysis. In areas with subacute to chronic necrosis, necrotic myocytes were infiltrated by histiocytes and

heterophils. Concurrent with degenerative myocytes were regenerative myocytes characterised by a decrease in
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myocyte diameter, hypertrophy of satellite cells, numerous vesiculated open nuclei, and sarcoplasmic basophilia

(Figure 1).

The endomysium adjacent to these myocyte changes was infiltrated by multifocal, mild to moderate numbers of

histiocytes with occasionally vacuolated cytoplasm (foam cells), heterophils, and lymphocytes. In these areas, as

well as more widespread, the endomysium and perimysium were variably mildly expanded by increased clear space

(edema), mild fibrosis, and mature adipose tissue. Muscle vasculature within the supporting stroma also variably

contained robust, perivascular (veins) to transmural infiltrates of lymphocytes and fewer other mononuclear cells. In

veins with transmural infiltrates, endothelial cells were frequently hypertrophied; lymphocytes preferentially

infiltrated and expanded the tunica intima; and rarely, there was replacement of the tunica media by eosinophilic,

smudged debris (vasculitis) (Figure 2).

Temporal progression of histologic lesions in the pectoralis major muscle

At 2 weeks of age, many muscle samples were largely unremarkable with minimal, peracute to acute, multifocal

myodegeneration of the pectoralis major muscle interpreted to be within normal limits. However, there were

commercial birds with robust lymphocytic perivascular to transmural vascular infiltrates and, variably the same or

different birds, exhibited more significant WBM, with severity scores for both lesions up to 3 (Figure 3A-C). Mild

lesions of myopathy and lymphocytic infiltrates including phlebitis were present in a few ACRB broilers. WBM

lesions in among all birds were most prevalent at 8 weeks (Figure 3 D-F). At all time points, WBM, excluding

mild (score 2) lesions, was polyphasic with concurrent degeneration and regeneration. Regeneration was more

prominent in tissue samples from older age groups.

Wooden breast myopathy and lymphocytic phlebitis severity scores significantly correlated with temporal

progression, with rs values suggestive of a moderate, positive correlation (Figure 4; rs =  0.47 and 0.51,

respectively; p  <  0.05). While a positive correlation was observed between WBM and lymphocytic phlebitis

severity scores (see Lymphocytic phlebitis section), individual samples occasionally had robust lymphocytic

phlebitis in the absence of robust WBM and vice versa.

Incubation temperature profile does not correlate with WBM score

Comparison of mean WBM scores between S and LH incubation temperature profiles by strain including all time

points as well as by sampling week revealed no significant, consistent differences (Figure 5). At week 2, the
 7

ACRB strain WBM scores significantly differed between the two incubation temperature conditions (p  =  0.0179),

but this difference was not present in any of the modern broiler strains and did not continue in subsequent weeks.

Sex does not correlate with WBM score

Comparison of WBM scores between male and female birds by strain including all time points as well as by week

revealed no significant differences (Figure 6, p  >  0.05).

Strain correlates with WBM score

When analysis was performed including all time points together and individually by sampling week, the ACRB

strain broilers had significantly lower WBM scores compared to all other broiler strains (p  <  0.05), with p  <  0.0001

at weeks 4, 6, and 8. None of the tissue sections from ACRB strain chickens received a WBM score >  2. In the

remaining strains, there was no significant difference when all time points were included together and uncommon,

inconsistent differences when compared by week. At weeks 4 and 6, A/A mean WBM scores were significantly

lower than strain C/C mean WBM score (p  =  0.0127 and 0.0196, respectively). This pattern was not present at week

2 and did not persist into week 8.

Body weight correlates with WBM score and is significantly different among strains
Spearman’s correlation coefficient rs between WBM score and body weight was calculated for each sampling week.

At 2, 4, 6, and 8 weeks of age, the rs value was significant (Figure 7, p  <  0.05), with values suggestive of a mild

positive correlation (rs =  0.29, 0.40, 0.41, and 0.47, respectively). Rs between WBM score and body weight was also

calculated across all sampling weeks but by strain and a similar, significant (p  <  0.05) positive correlation was

found, albeit slightly stronger represented by higher r s values, excluding the ACRB strain (rs= A/A 0.60; A/B 0.55;

C/C 0.64; ACRB 0.27).

In order to determine if differences in body weight between the strains was a confounding factor for interpreting an

association between WBM score and strain, mean body weight was compared among strains by week and sex. At

each sampling week, the ACRB chickens were significantly lighter than all other strains examined, with no

significant difference between sexes. At the final sampling week, there were no significant differences in mean body
 8

weight between A/A, A/B, and C/C strains, when compared within sex. At all sampling weeks excluding week 2,

males were significantly heavier than females within the same strain. Amongst males, there were no significant

differences in mean body weights among A/A, A/B, and C/C strains. Amongst females, at weeks 2, 4, and 6,

females from the C/C strain were significantly lighter than females from the A/B strain, though this difference did

not persist into the final sampling week.

Lymphocytic phlebitis score moderately to strongly correlates with WBM score

Spearman’s correlation coefficient rs value between WBM and lymphocytic severity scores within all sampling

weeks was significant (Figure 8, rs=0.68) and suggestive of a moderate to strong positive correlation.

Discussion
The pathogenesis of WBM in the pectoralis major muscle remains incompletely understood despite characterization

of its clinical, gross, histologic, and functional alterations. In this report, myodegeneration and regeneration in the

pectoralis major muscle was histomorphologically scored with sampling at multiple weeks. Scores were evaluated

against multiple variables to identify contributing factors and further expand the current understanding of WBM

pathophysiology.

Histologically, lesions consistent with the spectrum of WBM lesions were identified as early as 2 weeks in this

study. The severe form of WBM, called wooden or woody breast, is identified at slaughter without evidence of

antemortem clinical signs (Sihvo et al., 2014; Sihvo et al., 2017), though there have been recent, experimental

reports identifying evidence of WBM in chickens as young as 1 week of age (Papah et al., 2017; Sihvo et al., 2017).

WBMProgression of lesion severity throughout the grow-out period was present in this experimental flock,

identified both through positive Spearman correlation coefficient rs values, but more simply with the highest mean

scores of WBM severity at the final sampling point.

Lymphocytic phlebitis has been identified as a feature of WBM since its initial description (Sihvo et al., 2014), with

immunostaining characteristics of the lymphocytes consistent with T-cells (Sihvo et al., 2014). This is not a feature

of other myopathies in poultry, such as deep pectoral myopathy or vitamin E/ selenium deficiency induced

nutritional myopathy,yet it has only recently been further investigated (Kuttappan et al., 2016). Sihvo et al. (2017)

used histomorphologic scoring of the WBM and lymphocytic phlebitis components of WBM and found a strong

positive correlation between the two lesions. However, individual birds in which robust lymphocytic phlebitis was

present in the absence of WBM and vice versa were also identified. Papah et al. (2017) similarly identified
 9

lymphocytic phlebitis in WBM lesions, samples without gross evidence of WBM, and birds in the first week of life.

Our study confirms the strong positive correlation between muscle lesions and lymphocytic phlebitis in the

histologic evaluation of WBM lesions; the presence of lymphocytic phlebitis in chickens as young as 2 weeks of

age; and that individual muscle samples can have extensive lymphocytic phlebitis in the absence of significant

muscle lesions and vice versa.

This is the first study that also identifies lymphocytic phlebitis in a non-modern meat type chicken, the Athens

Canadian Random Bred strain, representative of 1950’s meat type chickens (Collins et al., 2016). While no WBM

severity score greater than 2 occurred in ACRB chickens, widespread and occasionally robust lymphocytic phlebitis

was identified within some of the muscle samples from this strain. The combination of these characteristics suggests

that lymphocytic phlebitis precedes and possibly contributes to the muscle injury through altered blood perfusion to

the region. Recently, Sihvo et al. (2018) used techniques to target the early pathogenesis of WBM lesions and

identified reduced vessel density and myofiber ultrastructural changes consistent with hypoxia in muscle adjacent to

WBM lesions, providing support that ischemia may play a role in WBM pathogenesis. Altered gene expression

suggestive of hypoxia and oxidative stress (Mutryn et al., 2015) also fits nicely into this picture of altered local

blood perfusion, though these changes could be a secondary response to the marked inflammation and edema.

Additional studies are needed and consideration should still be given that lymphocytic phlebitis is a secondary lesion

or possibly unrelated to WBM. Regardless, if lymphocytic phlebitis is a primary factor in the development of muscle

injury, then the remaining question is its aetiopathogenesis, with broad differentials to be considered including viral

and immune mediated causes (Sihvo et al., 2014).

The other strong positive correlation identified in this study was between WBM severity score and body weight,

with this correlation present when assessed both by strain and by week. Previous studies have identified this

correlation with WBM (Cruz et al., 2017) as well as in white striping myopathy (Lorenzi et al., 2014; Kuttappan et

al., 2012) and multiple other researchers have considered it to be at least a contributory factor in the disease (Sihvo

et al., 2014; Velleman and Clark 2015; de Brot et al 2016). The specific mechanism that results in this correlation

has been speculated to be insufficient capillary blood supply to remarkably hypertrophied myofibers (Joiner et al.,

2014; Sihvo et al., 2017), though any impact on the cardiovascular system, such as the previously described

lymphocytic phlebitis, within the context of pectoralis major muscle hypertrophy could play a role.
 10

While significant strain differences in WBM severity scores were identified in this study, this was limited to the

ACRB strain, with no differences identified among the 3 modern, commercial strains. ACRB strain birds were

significantly smaller than all other strains at all time points and, within the context of the correlation of WBM score

to body weight, it cannot be concluded that there is a clear genetic association with WBM severity score. Within the

past few years, multiple researchers have similarly evaluated strain differences in WBM, albeit through different

mechanisms, with none identifying a correlation (Bailey et al., 2015; Trocino et al., 2015; Radaelli et al., 2016).

Bailey et al. (2015) concluded from their analysis that greater than 90% of the variance in WBM results from

environmental and management factors rather than genetics.

The majority of myogenesis occurs in ovo (Joiner et al., 2014) and incubation temperature conditions have been

demonstrated to alter pectoralis major muscle meat quality traits and growth (Janisch et al., 2015), and

musculoskeletal leg health (Oviedo-Rondon et al., 2009). In this study, there was no significant difference in WBM

severity score between chickens hatched from standard or sub-optimal incubation temperature profiles. This

suggests environmental and management factors that contribute to WBM likely occur post-hatch.

In this study, no significant difference was identified in WBM severity scores between male and female birds.

Previously published reports on differences in WBM prevalence between sexes are contradictory. Trocino et al.

(2015) identified a lower prevalence of WBM in females compared to males, while Radaelli et al. (2016) did not

find the same to be true. Further evaluation of sex differences is needed.

In conclusion, this study identifies histologic evidence of WBM as early as 2 weeks of age with the prototypical

lesions identified, including myodegeneration, regeneration, fibrosis, inflammation, and lymphocytic phlebitis.

Positive correlations were identified between WBM severity scores, lymphocytic phlebitis scores, and growth rate.

The lymphocytic phlebitis was notably also present in the ACRB strain, most frequently without significant

myodegeneration, regeneration, or inflammation. While questions remain regarding the full pathogenesis of WBM,

the role of the lymphocytic phlebitis and vascular injury should be further explored as a central feature of the

disease.

Disclosure Statement
This work was supported in part by grant #691 from the US Poultry and Egg Association.

The authors of this publication report no financial or business activity that present a conflict of interest.

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Table 1. Bodyweight by strain per week (initially published in Chen et al., 2017)
Week
2 4 6 8
Strain Sex N Mean SEM Mean SEM Mean SEM Mean SEM
A/A F 68 506.9 ab 10.1 1588.6 cd 32.5 2950.0 bc 56.5 4161.7 b 64.1
M 68 545.7 a 7 .8 1756.3 ab 43.0 3439.7 a 77.7 4635.7 a 159.0

A/B F 66 528.4 a 11.2 1689.5 bc 28.7 3095.8 b 68.4 4144.4 b 62.1

M 71 531.9 a 9 .1 1838.7 a 27.5 3649.5 a 57.3 4831.4 a 173.5

C/C F 71 473.5 b 10.0 1482.4 d 28.7 2847.6 c 48.6 3893.4 b 78.8

M 66 506.6 ab 10.6 1759.7 ab 31.7 3478.8 a 62.8 5073.0 a 130.9

ACRB F 67 141.7 c 5 .5 329.3 e 9 .4 598.6 d 19.8 883.4 c 21.6

M 67 149.0 c 6 .9 394.1 e 10.7 718.9 d 36.1 1081.0 c 21.5
SEM – Standard error of the mean
abcd
Items that do not share superscript letters are significantly different (p < 0.05).
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Figure 1. Commercial broiler chickens, pectoralis major muscles, representative histomorphology severity scores.
Scores 1–3 are from 4 week birds; score 4 is from an 8 week bird. Score 1; normal breast muscle. Scores 2, 3, 4;
mild, moderate, and severe wooden breast myopathy. Score 4 muscle equates to ‘woody breast’. There is
myodegeneration and regeneration, necrosis, endo- and perimysial fibrosis, and inflammation. Small, basophilic,
irregular myofibers in score 3 are regenerating fibers. Loss of normal muscle architecture, fiber atrophy, splitting,
and vacuolation, and increased interstitial fibrosis are present in score 4 muscle.
Figure 2. Commercial broiler chickens, 4 weeks, pectoralis major muscles, lymphocytic phlebitis. A. Mild lesion.
Lymphocytes in the wall of the vein extend into the adjacent edematous perimysium. Endothelial cells are swollen.
Degenerating and regenerating myofibers are adjacent to the perimysium. Arteriole is unaffected. B. Moderate to
severe lesion. Wall of vein is densely infiltrated with lymphocytes. Intravascular lymphocytes adhere to the
endothelium amid swollen endothelial cells. C. Severe lesion. Wall of vein is densely infiltrated with lymphocytes.
Only a few remaining red cells identify the vessel lumen. A necrotic fiber is adjacent to the affected vein and
regenerating cells can be seen in the upper left corner. D. Severe lesion flanked by a basophilic, regenerating
myofiber with central rows of vesicular nuclei.
Figure 3. Commercial broiler chickens, 2 weeks, A-C; ACRB broiler chickens, 8 weeks, D-F, pectoralis major
muscles. Similar mild myopathic lesions occurred in both commercial and ACRB broilers at 2 and 8 weeks
respectively. A,D. Degeneration; B,E. Regeneration; C,F. Lymphocytic phlebitis. Although similar, muscle lesions
were less severe in ACRB broilers and did not progress in severity over time. Lymphocytic phlebitis lesions were
similar between Commercial and ACRB broilers except for number of affected birds and extent.
Figure 4. Mean WBM score (error bar  =  1 standard deviation) of strains by week. ACRB strain broilers had
significantly lower WBM scores compared to all other broiler strains across all weeks. At weeks 4 and 6, A/A mean
WBM scores were significantly lower than strain C/C mean WBM score; otherwise, there were no significant
differences among the commercial strains.
Figure 5. Mean WBM score (error bar  =  1 standard deviation) of both incubation temperature profiles by strain. No
significant differences were identified between the mean WBM scores of the 2 incubation temperature profiles in
any of the strains.
Figure 6. Mean WBM scores of both sexes by strain (error bar  =  1 standard deviation). No significant differences
were identified between mean WBM scores of male and female birds of the same strain.
Figure 7. Scatter plot of body weight by WBM score within a week. A mild positive correlation was identified
between WBM score and body weight with the correlation strengthening with temporal progression (r s: week 2  =
 0.29; week 4  =  0.40; week 6  =  0.41; and week 8  =  0.47)
Figure 8. Scatter plot of lymphocytic phlebitis score by WBM score for all birds. A moderate to strong positive
correlation was identified between WBM and lymphocytic severity score (r s=0.68).
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