A Case of Acute Pulmonary Edema, Splenomegaly, and Ascites in Guinea Fowl
Author(s): B. S. Cowen, H. Rothenbacher, L. D. Schwartz, M. O. Braune, R. L. Owen
Source: Avian Diseases, Vol. 32, No. 1 (Jan. - Mar., 1988), pp. 151-156
Published by: American Association of Avian Pathologists, Inc.
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AVIAN DISEASES32:151-156, 1988
A Case of Acute Pulmonary Edema, Splenomegaly,
and Ascites in Guinea Fowl
B. S. Cowen, H. Rothenbacher,L. D. Schwartz,A
M. O. Braune,and R. L. Owen
Departmentof VeterinaryScience, Collegeof Agriculture,
The PennsylvaniaState University, University Park,Pennsylvania16802
Received 7 August 1987
SUMMARY. Acutepulmonaryedema,splenomegaly, asciteswereobservedin a disease
outbreakin adult white and pearlguinea fowl. The clinical historyand gross and microscopic
lesions resembledthose describedfor marblespleendiseaseof pheasantsand avian adenovirus
groupII splenomegaly chickens.A small numberof intranuclear
of inclusionbodies werefound
in liver, spleen,andlungsectionsof affectedguineafowl.Attemptsto isolatevirusand serological
tests to detect the presenceof viral antigenswere unsuccessful.
Adult female pearl guinea fowl experimentallyexposed to pheasantand turkeyisolates of
type II avian adenovirusesdeveloped gross and microscopiclesions similar to those seen in
the field outbreak.The pheasantisolate was the more virulent. Intranuclear inclusion bodies
were observedin liver, spleen, and lung sections of pearlguinea fowl inoculatedwith either of
the virus isolates, and direct immunofluorescent examination revealed viral antigen in the
spleen and lung.
RESUMEN. Reportede Caso-Edema pulmonaragudo,esplenomegalia ascitisen gallinas
Se present6 un brote con edema pulmonaragudo, esplenomegaliay ascitis en gallinas de
Guinea adultas blancas y pintadas. La historia clinica y las lesiones macro y microsc6picas
fueronsemejantesa las observadasen la enfermedaddel bazo moteado de faisanesy la esple-
nomegaliacausadapor adenovirusaviar grupoII en pollos. Se encontr6 un nfimeropequenio
de cuerposde inclusi6nintranucleares seccionesde higado,bazo y pulm6n.No fructificaron
los intentosparaaislarel virus o paraidentificar antigenosviralespor medio de las pruebas
Gallinasde Guinea pintadasexpuestasexperimentalmente cepas del tipo II de adenovirus
de faisany pollo desarrollaronlesiones macroy microsc6picassimilaresa las observadasen el
brotede campo, siendo la cepa de faisanla mas virulenta.Se encontraron cuerposde inclusi6n
intranucleares seccionesde higado,bazo y pulm6nde gallinasde Guineapintadasinoculadas
con cualquierade las cepas y el examen por inmunofluorescencia directaidentific6el antigeno
viral en bazo y pulm6n.
A literature search failed to reveal any reports (HE) of turkeys are caused by type II avian ad-
of a guinea fowl disease resembling marble spleen enoviruses (1), and pheasants, chickens, and
disease (MSD) of pheasants (5) or avian ade- turkeys are the only known natural hosts (5).
novirus group II splenomegaly (AAS) of chickens However, fowl adenoviruses (type I avian ade-
(5). These diseases plus hemorrhagic enteritis noviruses) (1) have been isolated by Pascucci et
al. (15) from guinea fowl with pancreatitis.
No antibody to type II avian adenoviruses has
Paper No. 7512 in the JournalSeries of the Penn-
been detected in wild bird sera (2). Laboratory
ACurrentaddress:Animal Health Diagnostic Lab- studies have demonstrated that pheasant and
oratory, College of VeterinaryMedicine, P.O. Box chicken isolates of type II avian adenoviruses
30076, MichiganState University, East Lansing,MI will infect turkeys and that turkey isolates will
48909-7576. infect pheasants (3,6,7,12). Additionally, exper-
152 B. S. Cowen et al.
ltilmlllu.illill'l' ll6l/34 5rl""67
<, "-"-"~ ^,' -- '/,~ -,,, ~_DA'TE--
SPECIMENg :- e l
Fig. 1. Lungswith acute edema;enlargedand mottled spleens.
imental HE virus infection of golden pheasant, guinea fowl were observed to be affected. No
peafowl, chickens, and chukars produced en- vaccines or antibiotics had been administered,
larged and mottled spleens but no mortality (5). and no significant disease problems had been
The following is a case report of acute pul- encountered previously by either of these guinea
monary edema, splenomegaly, and ascites in adult fowl flocks.
guinea fowl. Results of experimental type II avi- Eight dead and four live guinea fowl hens were
an adenovirus infection of guinea fowl are also submitted to the Livestock and Poultry Veteri-
discussed. nary Diagnostic Laboratory at The Pennsylvania
State University for examination. Dead and sick
birds were cyanotic, and a greenish diarrhea was
observed on vent feathers. Several birds had dis-
An eastern Pennsylvania game farm experi- tended abdomens which contained copious
enced a sudden onset in mortality (33 of 1500; amounts of straw-colored fluid. Affected hens ap-
2.2%) among white guinea fowl breeders in late peared to be of normal weight and were ovulat-
April 1983. An egg-production drop of 30-50% ing, but the ovarian follicles were hemorrhagic,
was calculated for these breeders, which were and some had ruptured, causing egg-yolk peri-
maintained in four different houses. Collected tonitis. The lungs were edematous, and spleens
eggs were of normal size, shape, and color. Ap- were enlarged (2-4 x) and mottled (Fig. 1); le-
proximately 1 week following the disease onset, sions were similar to those described for MSD
pearl guinea fowl housed adjacent to the white and AAS (5). The lungs also exhibited acute
Pulmonaryedema, splenomegaly,and ascites in guinea fowl 153
Fig. 2. Lung (field case): A congested and edematous lung exhibiting a histiocyte with an amphophilic
inclusion body (arrow).H&E. 1000x.
congestion, and serosanguineous fluid exuded Bacterial culturing of liver sections, abdomi-
from the cut surfaces. Livers were swollen, and nal fluid, and swabs of the abdominal cavity of
some had areas of greenish discoloration on the both dead and moribund birds resulted in a mixed
surface. Most hens had a mild to moderate ca- flora of Escherichia coli, Staphylococcus epider-
tarrhal enteritis, which was not grossly hemor- midis, Streptococcusfaecalis, and Bacillus sp. Be-
rhagic. The intestinal walls of several hens were cause of the suspected viral nature of this disease,
thickened, and cecal worms were present. acute and convalescent sera were evaluated by
Histopathologic examination of hematoxylin- double immunodiffusion (DID) (4) and enzyme-
and-eosin (H&E)-stained sections of spleen linked immunosorbent assay (11) and were found
showed characteristic multifocal histiocytic hy- to be free of reovirus, reticuloendotheliosis virus
perplasia and necrosis. The pulmonary lesion was (REV), and type I and II avian adenovirus an-
acute congestion and edema (Fig. 2). Periportal tibodies (1,4). Spleens were examined for the
hepatitis and cholangitis with multifocal necrosis presence of type II avian adenovirus antigen by
was observed in the liver. Nuclear changes re- DID and direct immunofluorescence (DIF; fluo-
sembling those of type II avian adenovirus in- rescein isothiocyanate conjugate) (8) and were
fections (nuclear vesiculation and inclusion body also found to be negative. An attempt to isolate
formation) were seen in histiocytes of the spleen a type II avian adenovirus from spleen homog-
and lung (Fig. 2) and in hepatocytes (Fig. 3). The enates was made in a B-lymphoblastoid cell line
enteritis ranged from catarrhal to erosive-hem- (MDTC-RP19) (13,14) with no success. At-
orrhagic. Capillaria cross-sections were found in tempts to isolate other viruses by yolk-sac in-
the mucosa. Glomerular and tubular nephritis oculation ofspecific-pathogen-free (SPF) chicken
was present in the kidneys. No brain lesions were embryos (10) and inoculation of SPF chicken-
detected. Special staining techniques (Jenner- embryo-liver-cell culture (16) were also unsuc-
Giemsa and Warthin-Starry) for spirochetes re- cessful.
vealed nothing. In an effort to obtain additional information
A hematological evaluation of four affected about the etiology of the above described disease,
guinea fowl hens revealed a relative lymphope- three clinically normal 18-week-old pearl guinea
nia, monocytosis, and heterophilia. Red blood fowl hens were inoculated intravenously (0.5 ml)
cells had hypochromic nuclei and immature with approximately 5 x 104 TCID50 of pheasant
shapes. or turkey type II avian adenoviruses. Two hens
154 B. S. Cowen et al.
nuclearvesiculation(N). H&E. 1000x.
(representing one of each inoculum type) were and congested edematous lungs (pheasant virus
sacrificed at 4 days postinoculation for patho- only). Gross lesions were more marked in the
logical evaluation. No clinical signs were ob- pheasant virus (PV)-inoculated hen.
served before euthanasia. One hen-was held for Microscopic lesions were similar to those de-
assays of antibody stimulation. scribed for the reported field outbreak and were
Sacrificed hens exhibited distended gas-filled more intense in the guinea fowl exposed to the
intestines and ceca, enlarged marbled spleens, PV isolate. The spleen of the PV-inoculated fowl
q ?w !& V
inclusion bodies in pulmonaryhistiocytes (ar-
Fig. 4. Lung (experimental):
rows). H&E. 1000x.
Pulmonaryedema, splenomegaly,and ascites in guinea fowl 155
w d+lPa ;-
Fig. 5. Liver (experimental): inclusion body in a hepatocyte(arrow).Note the
nuclear vesiculation (N). H&E. 1000 x .
exhibited multifocal necrosis, and nuclei of his- DISCUSSION
tiocyteswere found to be vesiculatedand to con- Thegrossandmicroscopic lesionsin the spleens
tain amphophilic inclusion bodies. Acute and lungsof white and pearlguineafowl plus the
congestion and mononuclear pneumonitis was clinical history of this case are similar to those
detected in the lung of the PV-inoculatedfowl. describedfor MSD of pheasants(5) and AAS of
Pulmonary histiocyteshadvesiculatednucleiwith chickens (5).
eosinophilicto lavenderto slightlybasophilicin- The histopathologic changesseenin guineafowl
clusions (Fig. 4). Cholangiohepatitiswas seen in spleens (field case and experimental) included
the liver of the PV-inoculatedfowl. Numerous lymphocyticdepletion and proliferationand ne-
hepatocytes had enlarged nuclei with eosino- crosis of histiocytes.The type of intranuclear in-
philic to lavender inclusion bodies (Fig. 5). The clusion bodies observed in splenic histiocytes
pancreasof this bird had foci of hydropic de- have been describedas pathognomonicfor MSD
generation. of pheasants(18). Additionally,theseguineafowl
The histopathologic findings for the turkey- exhibited an acute pneumonic congestion and
virus-infected guineafowl weresimilarto but less edema, which closely resembledsigns described
intense than those found in the PV-inoculated for MSD and AAS (5).
guinea fowl; there was less necrosis and fewer Although no virus could be isolated and no
vesiculated nuclei of histiocytes with inclusion viral proteincould be detectedby DID and DIF
bodies. The microscopiclesions observed in the in guinea fowl spleens from the field outbreak,
intestine and kidney of experimentallyinfected the pathologicand DIF findingsof field and/or
guinea fowl (both viruses) were essentially the experimentalcases are suggestive of a "type II
same as those found in the field case. avian adenovirussplenomegaly"of guinea fowl.
Viralproteinwas detectedby DIF in the spleen and
The hematology renalhistopathology also are
and lung of experimentallyinfected guinea fowl suggestiveof a viral etiology. Since the existence
(both virus isolates), which supports the gross of virus and/or viral protein in type II avian-
and microscopicfindings.Weeklyassaysfor pre- adenovirus-infectedbirds (9,17) is transient, it
cipitin formation in guinea fowl experimentally is understandable that virus might be missed in
inoculated twice (3 weeks apart) with the PV field cases. Type II avian-adenovirus-induced
isolate were negativethrough3 months post pri- precipitincould not be detected in either field or
mary inoculation. experimentally infected guinea fowl. In retro-
156 B. S. Cowen et al.
spect, one additional virus-isolation technique 9. Gross, W. B., and C. H. Domermuth. Spleen
should have been attempted: the inoculation of lesions of hemorrhagic enteritis of turkeys. Avian Dis.
SPF turkey poults with the intestinal contents
and/or minced splenic tissue of dead or mori- 10. Hitchner, S. B. Virus propagation in embryo-
bund guinea fowl (4,6). nating eggs. In: Isolation and identification of avian
pathogens, 2nd ed. S. B. Hitchner, C. H. Domermuth,
More research is needed to clarify the etiology H. G. Purchase, and J. E. Williams, eds. Am. Assoc.
of the described guinea fowl disease. Avian Pathol., College Station, Texas. pp. 120-121.
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