The Diagnosis of Aspergillosis in Birds
Michael P.Jones, D VM, Dip.AB VI,
and Susan E. Orosz, PhD, DVM, Dip.ABVP
The diagnosis of aspergillosis can prove to be a difficult
challenge for the avian veterinarian. The diagnosis is
based on the cumulative findings resulting from a
thorough physical examination, appropriate hemato-
logic and chemistry panel analyte analysis, radiography,
endoscopy and laparoscopy, culture, and other ancillary
diagnostic tests. Despite many of the advances made in
diagnostic ability, the diagnosis is not as easy as it
seems. The current methods for the detection and
diagnosis of aspergillosis are reviewed.
Copyright 9 2000 by W. B. Saunders Company.
Key words: Avian aspergillosis, ELISA, antibody, anti-
gen, polymerase chain reaction, fungaL
spergiUus fumigatus is a ubiquitous, sapro-
A phytic fungus capable of causing severe and
life-threatening illness in birds and mammals,
including humans. Aspergillosis develops as a
result o f inhalation o f large numbers of spores
over a short period o f time or chronic exposure
to low levels of spores in a contaminated environ-
ment. Because of its ubiquity in the environ-
ment, it is generally accepted that patients with
aspergillosis may be concurrently affected by
some o t h e r disease process or stressor that inhib-
its the ability of their i m m u n e system to ward off
infection. Most clinically significant AspergiUus
infections are associated with Aspergillus fumiga-
tus, although A. flavus, A. nigeg, A. nidulans, and
A. terreus are also r e p o r t e d as pathogenic, albeit
less commonly. 1
Because of the route o f exposure, aspergillosis
typically affects the respiratory tract (trachea,
bronchi, lungs, and air sacs); however, infection
may b e c o m e invasive and secondarily affect the
coelomic cavity and central nervous system. 1 To
From the Universityof Tennessee, Collegeof VeterinaryMedicine,
Department of ComparativeMedicine, Knoxville, TN.
Address reprint requests to Michael P. Jones, DVM, Dip.
ABVP-Avian, Department of Comparative Medicine, College of
Veterinary Medicine, The University of Tennessee, PO Box 1071,
Knoxville, TN 37901-I-71.
Copyright9 2000 by W. B. Saunders Company.
1055-937X/00/0902-0001510. 00/0
doi:l O.1053/AX. 2000. 4619
52 Seminars in Avian and Exotic Pet Medicine, Vol 9, No 2 (April), 2000: pp 52-58
further complicate matters, clinical signs of infec-
tion with aspergillosis are not always attributable
to infection within the respiratory tract because
of dissemination of infection to other organ
systems. As a result, clinicians frequently rely
heavily on the use of diagnostic tests that attempt
to directly show the organism or serologically
show antigen material or antibody response to
infection. Ultimately, the diagnosis of aspergillo-
sis is based on cumulative findings obtained from
a t h o r o u g h history and physical examination,
appropriate hematologic and plasma biochemi-
cal analysis, radiography, endoscopic and laparo-
scopic evaluation, and other ancillary diagnostic
tests.
Clinical Examination
T h e diagnosis o f aspergillosis can be difficuit
if not frustrating at times, which may be in part
caused by differences in the pathogenicities of
isolates o f A. fumigatus and the status o f the
patient's i m m u n e system. 2 In humans, aspergillo-
sis is considered to be an immunologically medi-
ated lung disease and is commonly f o u n d in
i m m u n o c o m p r o m i s e d patients such as those
with acquired immunodeficiency syndrome, neu-
tropenic cancer, or transplanted o r g a n s ) ,4 T h e
same may hold true for avian patients in that
immunosuppressive events or situations play a
large part in the ability of the patient to fight off
infections with AspergiUus spp. Therefore, fea-
tures of the clinical examination (including a
t h o r o u g h history and physical examination) can
help the clinician to quickly and effectively arrive
at the correct diagnosis.
Environment
T h e environment in which a bird is kept can
provide key information that will help in deter-
mining a diagnosis. Many factors can act as
stressors to the bird's i m m u n e system, including
c o n c u r r e n t disease, trauma, toxicoses, recent
travel, and the administration of therapeutic
 Diagnosis of Aspergillosis
agents (in particular glucocorticoids) to n a m e a
few. Environmental factors that increase the
likelihood of infection with Aspergillus spp in-
clude high humidity or dampness as well as
excessive dryness and p o o r h u s b a n d r y and sani-
tation. I m p r o p e r l y discarded food may also pro-
m o t e the growth o f Aspergillus spp spores, as can
p o o r ventilation in enclosed areas. Spores are
often p r o d u c e d in h u m i d situations, b u t c a n n o t
sporulate when s u b m e r g e d in water. Sporulation
requires that the spores dry out for t h e m to
b e c o m e easily airborne. 5 An ideal e n v i r o n m e n t
for sporulation calls for a period of high humid-
ity, followed by a drying period. Inhalation of
airborne spores or conidia has been r e p o r t e d to
be the c o m m o n route of exposure. 6 These co-
nidia (2 to 3 p m in size) are capable o f bypassing
the physical barriers of the u p p e r respiratory
tract so that they deposit d e e p in the lung
p a r e n c h y m a a n d / o r the caudal air sac m e m -
branes. 7
O t h e r stressors c o m m o n to captive birds in-
clude reproductive activity, high temperatures,
and excessive h u m a n traffic t h r o u g h o u t exhibits
or cage areas. Free-ranging avian species may
also be stressed by similar environmental factors
occurring naturally, as well as by exposure to
environmental toxins such as oil c o n t a m i n a t i o n
or lead.
Signalment
Many clinicians consider certain species of
birds to be m o r e susceptible to aspergillosis than
others. C o m p a n i o n species such as African grey
parrots (Psittacus erithacus), blue-fronted Ama-
zon parrots (Amazona aestiva aestiva) and the
Indian hill m y n a h birds (Gracula religiosa) are
considered to have a higher prevalence o f asper-
gillosis. 8,9 O t h e r species t h o u g h t to be m o r e
susceptible include gyrfalcons (Falco rusticolus),
i m m a t u r e red-tailed hawks (Buteo jamaicensis),
goshawks (Accipiter gentilus), golden eagles (Aq-
uila chrysaetos), bald eagles (Haliaeetus leucocepha-
lus), rough-legged hawks (Buteo lagopus), pen-
guins, birds o f paradise, p h e a s a n t , a n d
waterfowl, s,9
Clinical Signs
Generally, aspergillosis appears as o n e of two
clinical conditions. T h e most c o m m o n is the
53
chronic f o r m in which a bird appears to go
through an i m m u n o c o m p r o m i s i n g event that
inhibits its ability to contain low-level infections
with AspergiUus spp spores, l~ T h e result is coloni-
zation of the respiratory tract with fungal granu-
lomas a p p e a r i n g in the trachea, syrinx, and air
sacs, especially in the caudal thoracic and abdomi-
nal air sacs. T h e second f o r m is an acute illness
that is seen m o r e frequently in cases o f p o o r
sanitation or h u s b a n d r y conditions allowing for
inhalation of a large n u m b e r of spores that
overwhelm the healthy i m m u n e system. T h e
result here is the rapid colonization of the
respiratory tract, in particular the lungs, which
often leads to severe dyspnea with death soon to
follow.
Clinical signs vary d e p e n d i n g on which f o r m
of the disease a bird develops. With the acute
form, clinical signs may develop over a few days
and can include dyspnea, lethargy, depression,
anorexia, weight loss, emaciation, polydipsia,
polyuria, exaggerated respiratory effort, cyano-
sis, and sudden death. 1~ Clinical signs seen with
the chronic f o r m can vary dramatically a n d may
include the same signs exhibited during an acute
infection. Additional clinical features of the
chronic f o r m may include a change in voice
character or quality, loss of voice, respiratory
stridor, ataxia, torticollis, seizures, unilateral or
bilateral rear limb paresis or paralysis, oculonasal
discharge, conjunctivitis, keratitis, nasal or perior-
bital swelling, beak malformations, exercise intol-
erance, and hepatomegaly, d e p e n d i n g on the
organ or organ systems affected. 1,s,1~
Diagnosis
As previously stated, the diagnosis of aspergil-
losis can be difficult at best, a n d no o n e test is
diagnostic for aspergillosis. This can be espe-
cially true of the chronic f o r m of the disease, in
which clinical signs can mimic m a n y o t h e r dis-
ease processes.
Hematology
Hematologic findings can be highly sugges-
tive of aspergillosis. Typically, birds that m o u n t
an a p p r o p r i a t e i m m u n e response show a hetero-
philic leucocytosis, monocytosis, lymphopenia,
hyperproteinemia, and a nonregenerative ane-
mia. T h e total white blood cell c o u n t (WBC) is
54- Jones and Orosz
usually higher than 20,000/pL and may range as
high as or higher than 100,000/pL. 1~ In those
patients with inadequate numbers of circulating
heterophils or inappropriate heterophil func-
tion, the WBC may be in the normal range or
significantly reduced. Morphologic changes such
as those seen with toxic heterophils can also be
associated with severe or deteriorating AspergiUus
spp infections. 13 Hyperproteinemia and nonre-
generative anemias are more consistently seen
with chronic forms of aspergillosis.
Biochemistry
Plasma biochemical analysis of patients sus-
pected of having aspergillosis can be rewarding
but not necessarily diagnostic of any particular
disease. Because o f the close proximity of the air
sacs with the peritoneal spaces and organs not
associated with the respiratory tract (liver, kid-
neys, and the reproductive and gastrointestinal
tracts), there lies the possibility of direct spread
of the disease from the respiratory tract to other
organ systems. Increases in aspartate aminotrans-
ferase (AST) and creatine kinase (CK), if pre-
sent, seem to be the most consistent findings.
Elevations in AST can result from extension o f
granulomas to the liver. Plasma bile acids may
help the practitioner in determining whether or
not elevations in AST are indicative of hepatic
disease and impaired hepatic function or originat-
ing from muscle damage. Elevations in CK may
result from muscle catabolism in emaciated pa-
tients or trauma resulting from seizures or over-
exertion. An increase in the globulins along with
a hyperproteinemia are other findings that may
also indicate possible infection with aspergillosis.
Cytology
Cytological examination of samples taken from
the respiratory tract can provide key information
toward achieving a diagnosis of aspergillosis.
Samples can be obtained from tracheal and air
sac washes, sinus aspirates, choanal swabs, impres-
sion smears from biopsy samples, or directly
from fungal granulomas. However, care must be
taken to prevent environmental contamination
of samples. If fluid is present in the coelomic
cavity, this can also be an excellent source in
identifying fungal organisms. A definitive diagno-
sis of aspergillosis can be made based on cytologi-
cal evidence of lesions consistent with aspergillo-
sis, including large numbers of heterophils,
macrophages, and multinucleated giant cells
(more often seen in a chronic inflammatory
response) in the presence of fungal elements. 14
Cytological samples stained with Wright's stain,
Diff-Quik stain (Baxter Healthcare Corporation,
Scientific Products Division, McGaw Park, IL), or
new methylene blue stain may reveal thick,
septate (and at times nonseptate) branching
fungal hyphae and conidiophores indicative of
aspergillosis.14
Radiology
Radiographs are one of the more helpful
diagnostic tools available for detecting aspergillo-
sis in avian species. They can especially be useful
in the latter stages of disease development.
Radiographs are also indicated when monitoring
disease progression and response to therapy.
Radiographic evidence of aspergillosis in early
stages of the disease may be difficult to identify.
However, lack of radiographic evidence of asper-
gillosis should not deter the veterinarian from
considering aspergillosis if other signs are consis- 9
tent with the disease. Minimally, lateral and
ventrodorsal views should be taken of the coelo-
mic cavity or other affected organ systems. In
patients that cannot handle an anesthetic epi-
sode, standing or perching lateral views as well as
a dorsoventral view can be quite helpful. How-
ever, it has been n o t e d that many patients actu-
ally do well u n d e r anesthesia because they are
receiving 100% oxygen and tend to breathe
more easily.
Radiographic signs consistent with aspergillo-
sis include a p r o m i n e n t parabronchial pattern;
loss o f definition or asymmetry of the air sacs;
hyperinflation of the abdominal air sacs; focal
soft tissue opacities within the oropharynx, peri-
orbital sinuses, trachea, syrinx, lungs, and air
sacs; thickening of the air sac membranes; outlin-
ing o f air sac membranes by fluid accumulation
within the coelomic cavity; and hepatomegaly or
renomegaly, s,1~ Unfortunately, when radio-
graphic signs are easily identifiable, the progno-
sis is very poor.
Endoscopy
Endoscopic evaluation, whether through the
use of flexible or rigid scopes, provides a wealth
 Diagnosis of Aspergillosis
of information that is often immediately avail-
able to the clinician. Because endoscopy allows
for visual inspection of an organ or o r g a n system,
it can provide additional diagnostic i n f o r m a t i o n
pertaining to the presence and severity of a
disease process as well as m o n i t o r i n g disease
progression or regression. Certainly, m o s t of the
clinical signs suggestive of aspergillosis can be
considered indications for endoscopic evalua-
tion of a patient; however, m a n y patients will
require at least a short anesthetic episode, which
may be risky in severely ill patients.
Standard a p p r o a c h e s to endoscopic evalua-
tion should be used to evaluate the entire respira-
tory tract, including the choanal opening, glot-
tis, trachea and syrinx, lung parenchyma, air
sacs, a n d the entire coelomic cavity. I f time a n d
the health of the patient permit, bilateral ap-
proaches to the coelomic cavity should be used.
T h e o r o p h a r y n x (especially a r o u n d the c h o a n a
and rima glottis) is easily evaluated for the
presence of granulomas or lesions suggestive o f
aspergillosis. The trachea should be evaluated in
its entirety to include the syrinx and p r i m a r y
b r o n c h i for plaques, granulomas, or o t h e r struc-
tural changes within the tracheal l u m e n or the
syrinx. This can be accomplished by use of a rigid
or a small flexible scope d e p e n d i n g on the size of
the patient examined. The lower respiratory
tract is best evaluated by laparoscopy for the
presence of granulomas within the lung paren-
c h y m a and on air sacs, thickening a n d cloudi-
ness of the air sacs (particularly the a b d o m i n a l
air sacs), and increased vascularization of the air
sac walls. In the latter stages of the disease, white,
yellow, gray, or green plaques can be visualized
lining the walls of the air sacs and within the lung
parenchyma.
Microbiology
Samples of tissues, fluids, a n d swabs should be
cultured for the presence o f Aspergillus spp.
However, it should be n o t e d that culture o f the
organism in the absence of lesions is n o t diagnos-
tic of aspergillosis because Aspergillus spp are
ubiquitous organisms. 15 Appropriate sources of
samples for culture include d e e p tracheal swabs,
culture of sinus aspirates, choanal swabs, a n d
material obtained f r o m endoscopic evaluation of
the coelomic cavity. Additionally, the tip of the
endoscope, provided sterile technique was used,
5~
can be an excellent source of material for culture
if swabbed immediately after removal. 16 Dee[
tracheal cultures can be obtained while th~
patient is awake or anesthetized. T h e swab s h o u k
be inserted as far as possible into the trachea
removed without contacting any other surfaces
swabbed on Sabouraud dextrose agar, a n d ther
cultured at 37~ (98.6~ for 18 to 24 hours. 9,r
Many fungal organisms including AspergiUus sp[
will grow on b l o o d agar; however, Sabourau(
dextrose agar is used specifically to inhibit bacte
rial proliferation and p r o m o t e fungal growth, a7,1~
Culture may also help to distinguish Aspergii
lus spp from o t h e r organisms such as Penicilliun
spp and Mucor spp, which may cause similal
lesions, s Small white colonies that can be cytologi
cally identified as AspergiUus spp can be presen
in as little as 18 hours and m a t u r e into the
characteristic blue-green colonies in 48 hours.'
In most cases, 1 to 4 colonies are required t(
make a definitive diagnosis of aspergillosis; how
ever, a single colony should not be r e g a r d e d a~
an incidental finding or culture c o n t a m i n a n t
especially in the presence of clinical signs sugges
tive of aspergillosis. 9
Electrophoresis
Plasma or s e r u m p r o t e i n electrophoresi:
(EPH) can be a valuable a n d reliable diagnostic
tool used to provide additional information con
cerning a patient's protein levels; however, it i~
only an adjunctive tool a n d should be used a~
such. EPH is especially helpful in obtaining
m o r e accurate d e t e r m i n a t i o n of the albumin
globulin (A-G) ratio than that provided by c h e m
istry panel analysis. 19 EPH is also helpful whet
m o n i t o r i n g disease progression and response t~
therapy. 19 T h e n o r m a l e l e c t r o p h o r e t o g r a m fol
birds has a p r e a l b u m i n fraction, followed b]
albumin (the largest protein fraction in health,
birds), alpha-1 and alpha-2 globulins, beta-1 anc
beta-2 globulins, and g a m m a globulins. Bird~
affected by mycotic diseases such as aspergillosi~
may show an increase in the beta globulins ir
acute stages. 19 Concomitantly, there is usually
decrease in the albumin concentrations, result
ing in a decreased A-G ratio. Chronic aspergillo
sis infections may show an increase in beta ol
g a m m a fractions or both. 19 Some chronicall,
infected birds may not m o u n t an appropriat~
i m m u n e response, resulting in h y p o p r o t e i n
56 Jones and Orosz
emia. In some patients with aspergillosis, it is not
unusual for protein concentrations to remain
within normal reference ranges. In these cases
shifts in protein fractions and resultant de-
creased A-G ratio as d e t e r m i n e d by EPH can
provide valuable information and may indicate
the need for further diagnostics.a9,2~
Serology
Several serological assays have been devel-
oped to assist in the diagnosis of aspergillosis.
These include the agar gel immunodiffusion
(AGID) test, the indirect enzyme-linked immuno-
sorbent assay (ELISA) for antibody, and the
ELISA for antigen. These tests are used fre-
quently; however, the results should be inter-
preted in conjunction with the clinical history
and other diagnostic tests.
AGID
The AGID technique measures precipitating
antibodies to A.fumigatus, and although sensitive
for long-standing disease, may not be able to
diagnose aspergillosis in its early stages. 2 Most
likely, the antibodies measured may represent
immunoglobulin (Ig) G levels. In a study on the
immunologic response o f aspergillosis in pi-
geons (Columba livia), the birds were immunized
by weekly injections of A. fumigatus. Using an
IgM- and IgG-specific A. fumigatus ELISA assay, it
was f o u n d that the IgM levels were detectable
early and reached their maximum levels by the
second week. In contrast, the IgG levels started
to increase only in the second week and reached
their maximum titer at 63 days (ninth week).21
Indirect ELISA for the Detection o f Antibody
The indirect ELISA (The Raptor Center, The
University of Minnesota, St. Paul, MN), in con-
trast to the AGID, has the ability to detect A.
fumigatus antigen as early as 1 week after expo-
sure to an infective n u m b e r of spores, including
those patients that are not showing overt signs of
aspergillosis. The test uses three species-specific
conjugates from turkeys, psittacines, and falcons
(primarily gyrfalcon and p e r e g r i n e falcon
plasma), and is believed to measure both IgM
and IgG (although this has not been confirmed)
(Redig PT, personal communication, March 29,
1999)9 The ELISA yields a positive or negative
cut-offvalue of 0.121 for the presence of antibod-
ies in the patient's plasma or serum. Titers less
than 0.121 are considered negative; patients with
values between 0.122 and 0.250 are considered
to have had exposure or low-grade disease with
p o o r antibody response, or to have recovered;
values between 0.251 and 0.500 are mid-range
positive and values greater than 0.501 are consid-
ered to be high positive. Values greater than
0.250 were always associated with active aspergil-
losis (Redig PT, personal communication, March
29, 1999). Species for which the antibody ELISA
has shown to work well include macaws, Amazon
parrots, African grey parrots, cockatiels (Nymphi-
cus hoUandicus), gyrfalcons, peregrine falcons
(Falco peregrinus), prairie falcons (Falco mexica-
nus), hybrid falcons, merlins (Falco columbm~us),
kestrel (Falco sparverius) , goshawks (Accipiter genti-
lis), cooper's hawks (Accipiter cooperi), cranes,
waterfowl, galliformes, penguins, and most diur-
nal raptors 9 (Redig PT, personal communica-
tion, March 29, 1999). The one known exception
is that n o n e of the conjugates were successful in
binding owl antibodies in the detection of asper-
gillosis. In o u r practice, this test tends to work
well in c o m p a n i o n birds with chronic aspergillus
infections. Once a diagnosis has been made, it
has been useful as a monitoring tool for the
bird's response to therapy. We often recheck the
plasma titers every 1 to 2 months during the
treatment phase.
A second antibody ELISA (The D e p a r t m e n t
of Comparative Pathology, Avian and Wildlife
Laboratory, University of Miami School of Medi-
cine, Miami, FL) is also available for the detec-
tion of antibody, and is based on techniques
developed at the University of Minnesota; how-
ever, this test uses different conjugates for anti-
body detection (Cray C, personal communica-
tion, February 19, 1999). This test yields a
positive or negative cut-off value of 1.4 for the
presence of antibodies in the patient's plasma or
serum. Antibody titers ranging from 0.0 to 1.3
are considered negative; values between 1.4 and
1.6 are considered to be weakly positive for the
presence of antibodies; a moderate positive value
is given for titers between 1.7 and 2.0, and
patients with titers greater than 2.0 are consid-
ered strongly positive (Cray C, personal commu-
nication, February 19, 1999).
Antibody detection systems are useful for
diagnosing and monitoring a patient's response
 Diagnosis of AspergiUosis
during therapy for aspergillosis; however, as with
many serological tests that measure antibody
production, several factors can limit the useful-
ness o f these tests. Some patients, d e p e n d i n g on
the severity of infection or the presence of o t h e r
disease (s), may not be able to m o u n t an appropri-
ate antibody response, resulting in the produc-
tion of low levels or no antibodies during an
infection (Cray C, personal communication, Feb-
ruary 19, 1999). Additionally, it is believed that
the location of the infection can result in limited
antigenic stimulation, and the stimulation of the
humoral i m m u n e system may be prolonged after
recent infection (Cray C, personal communica-
tion, February 19, 1999). Furthermore, antibody
levels can decline after remission of disease or
therapy 9 (Redig PT, personal communication,
March 29, 1999, and Cray C, personal communi-
cation, February 19, 1999). Thus, paired sam-
pling along with other diagnostic test results may
help in monitoring progression of disease and
determining whether inadequate antibody levels
warrant treatment.
Although both AGID and the ELISA tests for
antibodies report out titers, the level measured
in the patient does not necessarily correlate with
the clinical severity of the disease. A study in
Peking ducks (Anas platyrhynchos) and Cape
shelducks ( Tadorna cana) was unable to correlate
blood parameter values and increasing IgG-
specific ELISA titers for A. fumigatus. 22 This study
would support clinical observations that severity
of infection cannot be correlated reliably with an
antibody titer to AspergiUus spp. In a n o t h e r study
that involved turkey poults (Melaegris gallapavo),
AGID and ELISA levels seemed to be erratic and
did not correlate statistically when c o m p a r e d to
the severity of the pathological lesions. 2 A signifi-
cant difference was found, however, when low-
and high-mortality groups were c o m p a r e d with
mean ELISA and AGID results. 2
ELISA for the Detection of Antigen
T h e ELISA for the detection of Aspergillus spp
antigen is based on an assay developed in Europe
for the diagnosis of invasive aspergillosis in
h u m a n patients. The test measures antigen,
irrespective of its source, which also requires the
clinician to use clinical histories and other diag-
nostic tests to interpret the results. Patients with
environmental exposure to AspergiUus spp anti-
gen may have a weak positive test result. Addition-
57
ally, exogenous contamination o f plasma samples
may yield a positive result as well. Positive or
negative results should be j u d g e d with the knowl-
edge that the half-life of the antigen may not be
long and that the site of infection may also be a
factor (Cray C, personal communication, Febru-
ary 19, 1999). To this point, weak or negative
antigen results have been observed from birds
with confirmed tracheal aspergillus granulo-
mas. 2~ Finally, weak or negative antigen results
are often observed in the presence of positive
antibody titers. I m m u n e complexing may re-
move antigen from the sample, thus preventing
their quantitation in the ELISA assay. False posi-
tive results may also occur because this test is also
known to detect PeniciUium spp as well as other
AspergiUus spp (Cray C, personal communica-
tion, February 19, 1999).
Clinical Use o f Serological Tests
Clinically, the antigen and antibody tests are
useful in different situations. T h e authors con-
sider these tests to be reliable; however, different
clinical syndromes may present diagnostic chal-
lenges. Although the indirect ELISA can detect
antibodies as early as 7 to 10 days postinfection,
in subacute infections or in patients with a p o o r
humoral response (sequestered granulomatous
infections on air sac membranes, immunocom-
promised patients) testing for antigen may pro-
vide more information than testing for antibody
response alone. In more chronic cases in which
antigen levels may be low, ELISA for the detec-
tion of antibodies may be more helpful.
The Next Level
Despite the availability of the aforementioned
diagnostic tests, there is still a great deal of work
and investigation required to enhance the ability
to quickly and effectively diagnose aspergillosis
in avian species. In h u m a n medicine, diagnosti-
cians have been investigating the use of latex
agglutination (Pastorex AspergiUus latex aggluti-
nation test; Sandofi Diagnostics Pasteur, Marnes-
La-Coquette, France) and ELISA methods in
their efforts to detect AspergiUus spp antigen in
serum or urine by identifying galactomannan or
other carbohydrates. Unfortunately, these test
results can also be difficult to assess in patients
because false positives caused by cross-reactivity
with numerous fungi (A. flavus, A. nig~ A. terreus,
58 Jones and Orosz
Paecilomyces variotii, Penicillium s p p , Fusarium s p p ,
Rhizopus oryzae, Trichophyton s p p , Candida s p p ,
Geotrichum candidum, Saccharomyces cerevisiae, a n d
Cryptococcus neoformans) a n d b a c t e r i a l o r g a n i s m s
(Staphylococcus epidermidis, Enterococcus faecalis, Co-
rynebacterium jeikeium, Pseudomonas aeruginosa, a n d
Escherichia coli) m a y o c c u r . I n v a s i v e a s p e r g i l l o s i s
is a m a j o r c l i n i c a l c o n c e r n in i m m u n o c o m p r o -
m i s e d h u m a n p a t i e n t s b e c a u s e o f difficulty i n
detection. In severe systemic infections, investiga-
tors in h u m a n m e d i c i n e r e p o r t t h e u s e o f poly-
m e r a s e c h a i n r e a c t i o n ( P C R ) to d e t e c t A.fumiga-
tus a n d A. flavus i n h i g h - r i s k p a t i e n t s . 23
U n f o r t u n a t e l y , i n f e c t i o n w i t h o t h e r AspergiUus
s p p is n o t u n c o m m o n , w h i c h is l e a d i n g to t h e
d e v e l o p m e n t o f g e n u s - s p e c i f i c P C R assays to
d e t e c t all c l i n i c a l l y r e l e v a n t Aspergillus species. 23
I n a v i a n m e d i c i n e as e a r l y as 1996, r e s e a r c h e r s
investigated the development of a method for
i d e n t i f y i n g A. fumigatus D N A u s i n g a P C R i n
o s t r i c h e s , w i t h p r o m i s i n g results. 24 It is a d v a n c e -
m e n t s s u c h as t h e s e t h a t will e n h a n c e o u r d i a g n o s -
tic a b i l i t i e s to d e t e c t i n f e c t i o n w i t h AspergiUus s p p
( s p e c i f i c a l l y A. fumigatus) r a p i d l y a n d i m p r o v e
t h e l i k e l i h o o d o f a s u c c e s s f u l o u t c o m e in in-
fected birds.
Acknowledgements
T h e authors thank Drs. Carolyn Cray and Patrick
Redig for their assistance in the preparation of this
manuscript.
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