J. Comp. Path. 2020, Vol. 174, 104e112 Available online at www.sciencedirect.

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INFECTIOUS DISEASE

Cardiac Lesions of Natural and Experimental

Infection by Parrot Bornaviruses
J. Leal de Araujo*, S. S. Hameed†, I. Tizard‡, P. Escandon‡,
P. R. Giaretta‡, J. J. Heatleyx, S. Hoppes‡ and R. R. Rech‡
*Departamento de Ci^encias Veterinarias, Universidade Federal da Paraiba, Areia, Paraiba, Brazil, † College of Veterinary
Medicine, University of Baghdad, Baghdad, Iraq, ‡ Department of Veterinary Pathobiology and x Department of Small Animal
Clinical Sciences, Texas A&M University, College Station, Texas, USA

Summary
Lymphoplasmacytic inflammation associated with bornavirus N protein occurs in the epicardial ganglia,
myocardium and endocardium of birds diagnosed with proventricular dilatation disease (PDD). These path-
ological findings suggest that sudden death in psittacine birds might stem from cardiac compromise due to par-
rot bornavirus (PaBV) infection. Therefore, we investigated cardiac lesions in cases of PDD, searching
databases from 1988 to 2019, and reviewed three experimental studies of PaBV infection. Fifty cases of PDD
in birds infected naturally with PaBV and 27 cases of PDD in birds infected experimentally with PaBV (all
having descriptions of inflammatory cardiac lesions) were reviewed. For each case, five regions of the heart
were evaluated by light microscopy and immunohistochemistry (IHC). These regions were the epicardial
ganglia/nerves, the endocardium, the myocardium, the Purkinje fibres and the great vessels. Sudden death
was documented in 17/50 naturally infected cases, while 23/50 had digestive signs, and only 12/50 had neuro-
logical signs. Grossly, only five naturally-infected and five experimentally-infected cases had cardiomegaly or
hydropericardium. Epicardial ganglioneuritis was the most consistent microscopical finding in natural (46/50)
and experimental cases (26/27), followed by myocarditis (34/50) for naturally-infected and endocarditis for
experimentally-infected birds (6/27). PaBV-2 antigen was detected most frequently by IHC in the epicardial
ganglia (54/77) compared with the other tissues. This retrospective study demonstrates the presence of PaBV
protein and inflammation in the heart of birds infected with PaBV and suggests a link between PaBV and car-
diac disease and sudden death in psittacine birds.

Ó 2019 Elsevier Ltd. All rights reserved.

Keywords: epicardial ganglioneuritis; myocarditis; proventricular dilatation disease; sudden death

Introduction psittacine birds worldwide and is characterized by

Parrot bornaviruses (PaBVs) belong to the species
Pisttaciform 1 and 2 of the recently classified genus Or-
thobornavirus, which encompasses mammalian and
avian bornaviruses (Amarasinghe et al., 2018). In
birds, PaBV causes a lethal disease commonly
referred to as proventricular dilatation disease
(PDD) (Tizard et al., 2016; Amarasinghe et al.,
2018). This disease affects captive populations of
Correspondence to: R. R. Rech (e-mail: rrech@cvm.tamu.edu).
0021-9975/$ - see front matter
https://doi.org/10.1016/j.jcpa.2019.11.008
neurological and/or gastrointestinal signs (Hoppes
et al., 2010). Gross lesions are often limited to enlarge-
ment of the proventriculus and/or crop of affected
birds and emaciation (Hoppes et al., 2010; Tizard
et al., 2016). The most common microscopical
lesions are lymphoplasmacytic infiltration of the
central nervous system (CNS) and ganglia of
gastrointestinal (GI) organs such as the crop,
proventriculus, ventriculus and intestines. However,
inflammatory lesions in the ganglia and nerves of
the adrenal gland, eyes and heart have also been
Ó 2019 Elsevier Ltd. All rights reserved.
 Cardiac Lesions in Parrot Bornavirus Infection
described (Ouyang et al., 2009; Raghav et al., 2010;
Wunschmann et al., 2011; Payne et al., 2011b).
Avian autonomic cardiac innervation consists of
sympathetic nerves arising from the sympathetic
chain of the spinal cord; and parasympathetic nerves
arising from the vagus nerve (Dzialowski and
Crossley, 2015). After experimental intramuscular
inoculation in cockatiels (Nymphicus hollandicus),
PaBV has been demonstrated to travel first to the
CNS and then to peripheral organs such as the GI
tract and periadrenal ganglia. Occurrence of PaBV
in the vagus nerve has been detected as early as 35
days post inoculation (dpi), followed by the spread
to the heart at 40 dpi (Leal de Araujo et al., 2017b).
Several studies have reported cardiac lesions in
PaBV-infected psittacine birds and viral antigen has
been detected in the epicardial ganglia, the myocar-
dium and endocardium, which may be accompanied
of lymphoplasmacytic infiltrates (Ouyang et al., 2009;
Raghav et al., 2010). Additionally, sudden death has
also been reported in birds with PDD (Lierz, 2015).
Therefore, cardiac dysfunction in PDD could play a
role in the declining health of infected birds; however,
it is not clear how often the heart is affected in PDD.
The aim of this study was to evaluate cardiac lesions in
PDD and to determine whether PaBV nucleoprotein is
associated with these lesions in both naturally-infected
and experimentally-infected birds. We also reviewed
the clinical signs shown by these birds. We hypothesized
that PaBV antigen detected by immunohistochemistry
(IHC) and lymphoplasmacytic infiltrates would be pre-
sent in both natural and experimentally-induced PDD
and that sudden death would be a common clinical
sign associated with the presence of cardiac lesions.
Materials and Methods
Case Material
Natural cases were accessed from the avian pathology
archives of the Schubot Exotic Avian Health Center
(1988e1998) and the necropsy records from the
Department of Veterinary Pathobiology
(1999e2018), College of Veterinary Medicine and
Biomedical Sciences, Texas A&M University, Texas,
USA. Search terms included ‘PDD’, ‘proventricular
dilatation disease’, ‘macaw wasting disease’ and
‘avian borna virus’. Naturally occurring cases of
PDD were evaluated and filtered for histological de-
scriptions of lymphoplasmacytic infiltrates in the
heart. For inclusion in the study, cases must have
had evidence of classic lesions of PDD in the GI tract.
From these cases, histological slides of the heart were
re-evaluated to confirm the presence of inflammatory
lesions. Fifty cases of PDD with descriptions of cardiac
105
lesions were selected for additional evaluation based
on histological and immunohistochemical evaluation.
Necropsy accession histories were reviewed with
attention to species, age, sex and for client-reported
or clinician-reported clinical signs related to GI or
neurological dysfunction or sudden, acute or unex-
pected death. We equated the following terms with
sudden death: ‘sudden death’, ‘acute death’, ‘found
dead’, ‘death on examination’, ‘died shortly after pre-
sentation’, ‘sudden collapse’ or ‘died on the way to the
clinic’. We equated the following terms with wasting
(W): ‘wasting away’, ‘weight loss’, ‘progressive weight
loss, emaciation/cachexia’, which was included as a
GI sign. For neurological signs we included: ‘ataxia’,
‘falling off the perch’, ‘visual impairment’, ‘seizures’,
‘lethargy or depression’ and ‘weakness’.
Cockatiels used in three previous experimental
studies were also evaluated retrospectively for cardiac
lesions (Leal de Araujo et al., 2017b; Hameed et al.,
2018). These birds had been inoculated
intramuscularly with PaBV-2. Hearts of 27 cockatiels
with descriptions of cardiac lesions were re-evaluated
for gross and microscopical cardiac lesions, and the
presence of PaBV by IHC.
Histopathology and Immunohistochemistry
Serial transverse sections, from the great vessels to
apex, were obtained from the hearts (Fig. 1). Five car-
diac structures were evaluated by light microscopy:
the epicardial ganglia/nerve, the endocardium, the
myocardium, the Purkinje fibres and the great vessels.
All sections evaluated for inflammatory lesions were
also tested for PaBV N-protein by IHC. Briefly, IHC
was performed using polyclonal antibodies against a
specific region of the PaBV N-protein on serial tissue
sections, mounted on charged slides (Leal de Araujo
et al., 2017b; Hameed et al., 2018). Due to the
intranuclear replication site of PaBV, only cells with
intranuclear or intranuclear and intracytoplasmic
immunolabelling were considered positive, while
cells with only intracytoplasmic immunolabelling
were considered negative (Raghav et al., 2010). For
negative controls, a tissue section from a cockatiel
known to be positive for PaBV without the primary
antibody, and a tissue section from a cockatoo (Cacatua
moluccensis) known to be negative for PaBV and treated
with the primary antibody were used.
Reverse Transcriptase Polymerase Chain Reaction
Cardiac tissues from birds in experimental studies one
and three were analysed retrospectively for by reverse
transcriptase polymerase chain reaction (RT-PCR)
targeting the PaBV matrix (M) protein gene and
106 J. Leal de Araujo et al.

the phosphoprotein (P) gene (Guo et al., 2014;
Hameed et al., 2018). We also investigated PCR
results from study 2, where cardiac tissues were
tested in duplicate for expression of the genes
encoding the M and the P proteins. Briefly, mRNA
encoding the PaBV P protein was detected by using
a Taqman RT-PCR assay performed with
TaqManÒ Fast Virus 1-Step Master Mix (Invitro-
gen, Carlsbad, California, USA) and PaBV P
primers: 50 AAGAAGAA[Y]CC[Y]TCCAT-
GATCTC-3 and 50 -AA[Y]TGCCGAAT[B]A[R]
GTCATC-30 and a Taqman probe (50 -FAM-TCGA-
TAACTG [Y]TCCCTTCCGGTC-BHQ-30 ).
mRNA encoding the PaBV M protein was detected
by using PaBV M primers: 50 -
GGTAATTGTTCCTGGATGG-30 and 50 -ACAC-
CAATGTTCCGAAGACG-30 , and a Taqman probe
(50 -FAM-TCGATAACTG [Y]
TCCCTTCCGGTC-BHQ-30 ).
Results
Clinical Data
The clinical history of the 50 naturally-occurring
cases (Table 1) showed that 31 birds had signs of GI
tract dysfunction, including emaciation, anorexia,
regurgitation, diarrhoea or undigested seeds in the
faeces. Only 18 birds had neurological signs, which
included ataxia, lethargy, depression and seizures,
and 13 birds had both GI and neurological signs.

Fig. 1. Sampling technique of the heart for this study. (A) Serial sections were collected from the great vessels of the heart to the apex,
beginning with a transverse section to include both ventricles (B). This transverse section (C) facilitates the visualization of multiple
epicardial ganglia (inset) on histological evaluation, especially in the hearts of the larger species (macaws, cockatoos, African grey
parrots).
 Cardiac Lesions in Parrot Bornavirus Infection 107

Table 1
Common clinical signs of psittacine species with naturally occurring proventricular dilatation disease

Bird number Genus Sex Age Sudden death GI signs Neurological signs

1 Cockatoo (Cacatua sp.) F 8 years e W LD

2 Cockatoo (Cacatua sp.) F 10 years e W WE
3 Cockatoo (Cacatua sp.) F NI SD e e
4 Cockatoo (Cacatua sp.) NI 8 weeks e R WE
5 Cockatoo (Cacatua sp.) F NI SD R e
6 Cockatoo (Cacatua sp.) NI 1.5 years e W e
7 Cockatoo (Cacatua sp.) M 30 years e A, W e
8 Cockatoo (Cacatua sp.) NI 7 months e W, D, R e
9 Cockatoo (Cacatua sp.) NI 5 weeks SD R e
10 Cockatoo (Cacatua sp.) F 17 years e R e
11 Cockatoo (Cacatua sp.) F 4 months e WA, R LD
12 Macaw (Ara sp.) M 5 months e R, W e
13 Macaw (Ara sp.) NI 1 year SD e e
14 Macaw (Ara sp.) F 20 weeks e A, CSD e
15 Macaw (Ara sp.) M 15 years e V,D L
16 Macaw (Ara sp.) F 7 months e R LD, S
17 Macaw (Ara sp.) M 4 years e W, A e
18 Macaw (Ara sp.) M NI SD e e
19 Macaw (Ara sp.) F NI SD e e
20 Macaw (Ara sp.) F 8.5 years e A, USF LD
21 Macaw (Ara sp.) NI 8 months e W,D,R e
22 Macaw (Ara sp.) M 7 months SD R e
23 Macaw (Ara sp.) NI 8 years e R, CSD LD
24 Macaw (Ara sp.) NI 21 years SD e e
25 Macaw (Ara sp.) NI 2.5 years e e e
26 Macaw (Ara sp.) NI 38 years SD e FDB
27 Psittacus sp. F 5 months e R, W L
28 Psittacus sp. F 4 years e W AT
29 Psittacus sp. NI <2 years SD A WE
30 Psittacus sp. F 5 years e W e
31 Psittacus sp. F NI e e S
32 Psittacus sp. F NI e A, W, USF WE
33 Psittacus sp. M NI SD e e
34 Psittacus sp. M 2 years SD W e
35 Psittacus sp. M 5 months e W e
36 Psittacus sp. F NI SD e e
37 Psittacus sp. M 6 months e R L
38 Psittacus sp. M 12 years e e AT, VI, FDB
39 Psittacus sp. M NI e e FDB
40 Psittacus sp. M NI e e e
41 Alisterus sp. F 1 years SD e e
42 Aratinga sp. M <1 year e W e
43 Aratinga sp. F NI SD e e
44 Guarouba sp. M 4 years SD e e
45 Guarouba sp. F 1.5 years e W, USF e
46 Thectocercus sp. M 2 years e A, R e
47 Myopsitta sp. NI 4 weeks SD e e
48 Myopsitta sp. F 15 years e e L
49 Nymphycus sp. F 5 years e W e
50 Poicephalus sp. NI NI e e e

F, female; M, male; NI, no information; SD, sudden death; GI, gastrointestinal; W, wasting; R, regurgitation, AN, anorexia; D, diarrhoea; CSD,
crop stasis or distension; V, vomiting; USF, undigested seeds in faeces; WE, weakness; AT, ataxia/falling off perch; VI, visual impairment; LD,
lethargy or depression; FDB, feather-damaging behaviour; S, seizures.
108 J. Leal de Araujo et al.
Sudden death was described in 17 cases (34%), while
feather damaging behaviour was reported in three
cases. An echocardiogram of a naturally-occurring
case of PDD revealed evidence of tachycardia (i.e.
heart rate of 375 beats/min post sedation) with occa-
sional arrhythmia, pericardial effusion, ventricular
dilation, decreased ventricular fractional shortening
(20%) and mitral regurgitation, suggestive of cardiac
insufficiency.
The African grey parrot (Psittacus erithacus) was the
most frequent species (14 individuals) and also the
species to most often present with sudden death (five
cases), followed by the blue and gold macaw (Ara ara-
rauna) (five individuals and one case of sudden death).
In experimental study 1, one cockatiel presented
with acute signs of depression and lethargy and died
shortly thereafter. Another was found dead without
any prior signs of illness. In study 3, all eight cocka-
tiels presented with classical GI signs of PDD,
including weight loss and undigested seeds in the
faeces.
Out of the 50 birds, 21 were female and 16 were
male; however, sex was not recorded for 13 cases.
Gross Lesions
Gross cardiac lesions were found in five naturally-
infected cases. Cardiomegaly was observed in three
cases, including a PaBV-positive Quaker parrot (My-
opsita monachus) that was presented with an enlarged
right atrium (Fig. 2A), and an African grey parrot
with cardiomegaly. Hydropericardium was reported
in two cases. In the experimentally-infected cases,
cardiomegaly (Fig. 2B) was seen in four birds and hy-
Fig. 2. Gross cardiac lesions in psittacine birds with PDD were uncommon. Heart sections from three different cases are shown. An
enlarged right atrium was apparent in a natural (A; photograph courtesy of Dr. L. Bryan) and an experimental case (B), and hy-
dropericardium was documented in one experimental case (C).
dropericardium (Fig. 2C) was documented in one
case.
Histopathology and Immunohistochemistry
Microscopical lesions and PaBV immunolabelling
showed that 95% of the naturally-infected cases of
PDD with cardiac lesions had lymphoplasmacytic
infiltration of the ganglia (see Fig. 3A) and nerves of
the epicardium (Table 2). Myocarditis (Fig. 3C)
was present in 68% of these birds, while Purkinje fi-
bres were effaced by lymphoplasmacytic infiltrates
in only 36% of the cases. Mural endocarditis was
observed in 20% of all cases of PPD with cardiac le-
sions. Eight cases had ‘pan-carditis’. Additionally,
18% of the cases had lymphoplasmacytic infiltration
of the tunica media of the great cardiac vessels. No in-
flammatory lesions were observed in the valves.
Of the 46 cases of epicardial ganglioneuritis, 30
were also positive for PaBV-2 IHC (Fig. 3B), while
of the 34 cases with myocarditis, 20 had positive im-
munolabelling (Fig. 3D). For the endocardium and
Purkinje fibres, only 20% and 23%, respectively, of
the cases with inflammation in these areas were also
positive for PaBV-2. For the tunica media of the
great vessels, although only nine cases had
perivascular lymphoplasmacytic infiltrates, PaBV-2
antigen was detected in 15 cases. All naturally-
infected African grey parrots had ganglioneuritis in
the epicardium and eight of the 14 African grey par-
rots also had myocarditis. Four of five blue and gold
macaws had ganglioneuritis in the epicardium or
myocarditis.
 Cardiac Lesions in Parrot Bornavirus Infection 109

Fig. 3. Histological inflammatory lesions in the heart of psittacine birds with PDD. (A) Epicardial ganglioneuritis was the most frequent
histological finding and the cellular infiltrate was comprised mostly of lymphocytes and plasma cells. HE. Bar, 20 mm. (B) PaBV-2
antigen in the nucleus and cytoplasm of neurons and inflammatory cells. IHC. Bar, 20 mm. (C) Lymphoplasmacytic myocarditis.
HE. Bar, 50 mm. (D) Inflammatory cells and cardiomyocytes immunolabelled for PaBV-2. IHC. Bar, 50 mm.

In experimental study 1, 12 cockatiels had lympho-
plasmacytic infiltration of the heart, but had lympho-
plasmacytic infiltration of the nerves and/or
epicardial ganglia. Lymphoplasmacytic infiltrates in
the myocardium and endocardium were present in
34% of the cockatiels. No inflammatory lesions were
observed in the Purkinje fibres or great vessels.
In experimental study 2, seven cockatiels had lym-
phoplasmacytic infiltration of at least one of the five
regions evaluated. Six cockatiels (85.7%) had lym-
phoplasmacytic inflammation of the ganglia and/or
nerves of the epicardium. This type of inflammation
also occurred in the myocardium and Purkinje fibres
of two cockatiels. No inflammatory lesions were pre-
sent in the myocardium or endocardium of the cock-
atiels from this study.
In experimental study 3, all eight cockatiels devel-
oped lymphoplasmacytic infiltration of the heart with
further such inflammation in the ganglia and/or
nerves of the epicardium. These infiltrates were also

Table 2
Psittacine birds with inflammatory lesions and positive bornavirus antigen immunolabelling in the heart

Epicardial ganglia Myocardium Endocardium Purkinje fibres Great vessels

Natural cases (n ¼ 50)

HE 46 34 10 17 9
IHC 30 20 2 4 15
Experimental study I (n ¼ 12)
HE 12 4 4 0 0
IHC 10 4 1 0 2
Experimental study II (n ¼ 7)
HE 6 0 1 1 0
IHC 6 0 1 1 0
Experimental study III (n ¼ 8)
HE 8 1 1 0 0
IHC 8 1 1 0 0
110 J. Leal de Araujo et al.
observed in the myocardium of one cockatiel and in
the endocardium of another. No inflammatory lesions
were observed in the Purkinje fibres or great vessels.
For the experimental studies, only the first study
had differences between the number of birds with in-
flammatory lesions and the number of birds with pos-
itive IHC. Ten birds were positive for PaBV
immunolabelling of the epicardial ganglia, while
four cases were positive for the myocardium, one
case was positive for the endocardium and two cases
were positive for the great vessels. No immunolabel-
ling was detected in the Purkinje fibres. In the other
two experimental studies, immunolabelling was pre-
sent in all regions with inflammation.
Reverse Transcriptase Polymerase Chain Reaction
All 27 samples from the experimental studies tested by
RT-PCR were positive for expression of PaBV genes.
Discussion
We retrospectively analysed cardiac lesions in 77 psit-
tacine birds that were infected with PaBV and/or had
a diagnosis of PDD, including both natural and
experimental infections, over a period of 31 years
from two different necropsy databases and tissue
banks. Out of 50 naturally-infected cases, 34 dated
from before PaBV was recognized as the causative
agent of PDD (Honkavuori et al., 2008; Kistler et al.,
2008). In these cases, IHC was used to confirm
PaBV as the cause of ‘macaw wasting syndrome’.
We identified lymphoplasmacytic infiltration of the
epicardium, myocardium, endocardium, Purkinje fi-
bres and/or cardiac great vessels. These results asso-
ciate inflammatory lesions of the heart in psittacine
birds with PDD, as described elsewhere (Raghav
et al., 2010; Heffels-Redmann et al., 2011; Payne
et al., 2011a; Leal de Araujo et al., 2017a). In the 27
experimentally-infected cases, epicardial ganglio-
neuritis was the most consistent finding, representing
97% of the cardiac lesions. Some of the
experimentally-infected cockatiels lived as few as
114 dpi, which might not have been sufficient time
for PaBV to spread to other regions of the heart.
There is individual and interspecific variation, but
generally, epicardial ganglia in avian species are
concentrated in the dorsal areas, over the posterior
atrial walls and near the great vessels of the heart
(Smith, 1971).
Cardiac diseases in pet and wild birds may be due
to infectious, congenital, toxic, neoplastic or idio-
pathic causes. Two studies have systematically
searched for cardiac lesions in psittacine birds submit-
ted for necropsy examination. In one study, 26
(9.7%) of 269 psittacines subjected to necropsy exam-
ination had evidence of cardiac disease (Oglesbee and
Oglesbee, 1998). In another study of 107 psittacines
submitted for necropsy examination, 36% had gross
lesions in the heart, great vessels of the heart, or
both (Krautwald-Junghanns et al., 2004). The main
histological findings reported for these birds included
myocarditis (59.8%), focal coagulative necrosis
(23.4%) and lipomatosis cordis (48.6%). However,
these studies were published before PaBV was known
to be the causative agent of PDD. We found a reason-
able incidence of myocarditis; >10%, or seven of 50
cases of naturally-occurring PDD. We suggest that
the lack of myocarditis in experimentally-infected
cases may have been due to the relatively short exper-
imental time (generally <200 days), innate natural
resistance of the species used and the bornavirus geno-
type or entry route of naturally-infected cases, which
remains relatively uninvestigated.
Although not frequently reported in pet birds, cardio-
megaly was observed in our study. Hypertrophic and
dilated cardiomyopathy are described uncommonly in
parrots (Schmidt et al., 2015). In Mynah birds (Acrido-
theres spp.), cardiomegaly is reported more commonly
and is associated with haemochromatosis (Ensley et al.,
1979; Rosenthal and Stamoulis, 1993). Dilated
cardiomyopathy in young turkeys, also known as
‘round heart disease’, frequently causes cardiomegaly
and sudden death (Beaufrere and Wakamatsu, 2014).
Although described in chickens (Gallus domesticus), round
heart disease is considered rare in this species (Julian,
2005). In one study evaluating congestive heart failure
in African grey parrots, identification of the aetiology of
cardiac disease was not performed; however, one of the
parrots suffered from advanced PDD, with cardiomegaly
and lymphoplasmacytic infiltrates in the myocardium
and epicardial ganglia found on post-mortem examina-
tion (Juan-Salles et al., 2011). The number of African
grey parrots positive for PaBV with cardiac lesions in
this study suggests that some psittacine species may be
more likely to develop inflammatory cardiac lesions
than others. However, our study numbers were small,
with similar percentages of macaws (5/13) and cockatoos
(3/11) presenting with sudden death.
PaBV-2 antigen was not detected by IHC in the
heart of all naturally-infected cases with inflamma-
tory lesions. There are two possible explanations
for this observation, either or both of which may
have contributed to this finding. Firstly, some of
the formalin-fixed and paraffin wax-embedded
(FFPE) tissues used in this study were collected
over 30 years ago, which might have interfered
with or otherwise reduced the immunoreactivity of
the sections via a phenomenon known as antigen
decay (Grillo et al., 2015). Secondly, other PaBV
 Cardiac Lesions in Parrot Bornavirus Infection
genotypes (PaBV genetic diversity) could have been
involved in some of these cases, leading to lack of
cross-reaction with the antibody used in the study.
Experimentally, different PaBV genotypes likely
associate with different clinical manifestations
(Piepenbring et al., 2016), with clinical signs of either
a digestive or neurological nature. Some genotypes
of PaBV could have a more pronounced cardiac
tropism, but more study is needed to determine
this. Interestingly, PaBV-2 antigen was detected in
the tunica media of great vessels in six naturally-
infected cases and two experimentally-infected cases,
which lacked perivascular cuffing or vasculitis.
It might be suggested that the lack of PaBV immu-
noreactivity in some cardiac inflammatory lesions
supports the hypothesis that another infectious agent
is responsible for PaBV/PDD; however, all of the pre-
sent cases were diagnosed as PDD positive via histopa-
thology. For future studies we recommend that
genotype determination is included so that clinical,
gross and histopathogical changes and genotype asso-
ciations may be investigated further.
In cell culture, PaBV is non-cytopathic and the
behaviour of the virus in vivo appears similar
(Tizard et al., 2016; Murray et al., 2017). Injury to
the tissues is likely caused by T-cell infiltration,
rather than a virus-induced lesion (Hameed et al.,
2018). As seen in the present study, PaBV can be de-
tected in tissues without associated inflammatory or
circulatory lesions. More immunopathological studies
are required to clarify the correlation between PaBV
infection and gross, histopathological and clinical
manifestations of disease.
This retrospective study supports the hypothesis that
PaBV may be involved in the development of cardiac
disease in psittacine birds, and that PDD must be
included as a differential diagnosis in cases of sudden
death or when the necropsy examination reveals car-
diac alterations such as cardiomegaly or hydropericar-
dium. Because epicardial ganglioneuritis can be
considered a diagnostic histological lesion of PDD in
both natural and experimental cases, serial sections of
the heart between the great vessels and the level of the
atrioventricular valves are recommended for routine
post-mortem screening of birds affected by sudden
death or those with clinical signs suggested of PDD.
Acknowledgments
The authors thank D. Turner and the staff of the Vet-
erinary Pathobiology Histology Laboratory for tech-
nical support throughout this study, and the
Schubot Center for Avian Heath for funding this proj-
ect.
111
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½ Received,
Accepted, November 26th, 2019 
July 9th, 2019